A kind of white flesh ganoderma lucidum standard medicinal material extract and its detection method and use

Abstract

The present invention provides a white-fleshed Ganoderma lucidum standard medicinal material extract. The present invention provides a method for detecting white-fleshed Ganoderma lucidum standard medicinal material or its extract. The present invention also provides a method for identifying the authenticity of white-fleshed Ganoderma lucidum medicinal material, as well as a method for distinguishing between cultivated and wild white-fleshed Ganoderma lucidum. The present invention establishes a white-fleshed Ganoderma lucidum fingerprint and performs chemometric analysis to achieve authenticity identification of white-fleshed Ganoderma lucidum, as well as identification of cultivated and wild white-fleshed Ganoderma lucidum. The detection method of the present invention has strong repeatability, good precision, stability, and strong operability, and can distinguish the authenticity of white-fleshed Ganoderma lucidum medicinal material, as well as cultivated and wild white-fleshed Ganoderma lucidum varieties, with high accuracy.

Description

Technical Field

[0001] The invention relates to a white-fleshed ganoderma lucidum standard medicinal material extract, a detection method and application thereof. Background Art

[0002] Ganoderma lucidum is a large fungus belonging to the genus Ganoderma of the Polyporaceae family and is a precious traditional Chinese medicine in my country. There are many varieties of Ganoderma lucidum, with more than 80 species identified so far. Among them, Ganoderma sinense Zhao, Xu et Zhang and Ganoderma lucidum (Leyss.ex Fr.) Karst. are included in the 2020 edition of the "Chinese Pharmacopoeia". They have the effects of replenishing qi and calming the mind, relieving cough and relieving asthma. Ganoderma is produced throughout the country, including Zhejiang Province, Shandong Province, Heilongjiang Province, Yunnan Province and other places. The main active ingredients of Ganoderma lucidum are polysaccharides, triterpenes, etc. In addition, Ganoderma lucidum also contains proteins, amino acids, polypeptides and other ingredients. Modern pharmacological studies have shown that Ganoderma lucidum has anti-tumor, immune regulation, anti-aging, liver protection, and cardiovascular regulation effects.

[0003] White Ganoderma (Ganoderma leucocontextum), a prominent species of the genus Ganoderma in southwestern my country, is primarily found in Yunnan and Tibet and has been cultivated artificially. While research on Ganoderma lucidum and Ganoderma rubra is extensive and in-depth, research on white Ganoderma lucidum is limited, focusing primarily on biological identification, chemical composition, and neuroprotective effects. Pan Jun et al., "Analysis of Chemical Composition and In Vitro Antioxidant Activity of White Ganoderma," Food Industry Science and Technology, Vol. 42, No. 9, reported that white Ganoderma lucidum is rich in nutrients. Component analysis revealed crude protein content of 18.02% to 19.02% and crude fiber content of 14.30% to 20.05%. The main active ingredients in white Ganoderma lucidum, polysaccharides, triterpenes, and ganoderic acid A, are present in amounts of 0.99% to 1.42%, 1.22% to 1.40%, and 0.069% to 0.084%, respectively, significantly higher than those in Ganoderma lucidum and Ganoderma rubra. The total amino acid content of white Ganoderma lucidum ranges from 113.61 to 163.51 mg / g, with glutamic acid and aspartic acid at higher levels, at 16.12 to 22.62 mg / g and 12.02 to 15.61 mg / g, respectively. The content of medicinal amino acids in white Ganoderma lucidum exceeds 50% of the total amino acid content. White Ganoderma lucidum is characterized by high levels of calcium, magnesium, and iron, making it nutritionally valuable. Its magnesium, calcium, and iron contents range from 914.25 to 942.56 mg / kg, 575.18 to 807.25 mg / kg, and 380.00 to 465.35 mg / kg, respectively. The polysaccharide and triterpenoid content of white Ganoderma lucidum is higher than that of other Ganoderma lucidum varieties, and white Ganoderma lucidum contains novel polysaccharides and triterpenes. Ganoderma polysaccharides, the primary active ingredient in Ganoderma lucidum, can be isolated from its mycelium, spores, and fruiting bodies. Ganoderma polysaccharides have a complex structure and possess potent pharmacological properties, influenced by their branching and side chains. There are many types of terpenoid compounds in Ganoderma lucidum, which have strong medicinal properties and can be used to lower blood sugar, fight cancer, etc. In addition, a large amount of nucleotides and sterols in white-fleshed Ganoderma lucidum also have certain medicinal value and play an indispensable role in maintaining the body's physiological functions (Qiu Hao, et al., Research Progress on Active Ingredients and Their Biological Activities of White-fleshed Ganoderma Lucidum, Guizhou Science 39(3): 2021).

[0004] Currently, there is little research on the quality standards of white-fleshed Ganoderma lucidum. Pan Jun et al., "Heavy Metal Content and Toxicity Evaluation of White-fleshed Ganoderma Lucidum from Different Origins," Journal of Southwest Agriculture, Vol. 34, No. 11, 2021, monitored the changes in heavy metal content in white-fleshed Ganoderma lucidum from different origins in Yunnan and evaluated its safety. Application number CN202210732360.5, Invention Name: Specific Molecular Markers and Methods for Identifying White-fleshed Ganoderma Lucidum, Yunbai Ganoderma Lucidum Variety 2. By using the specific molecular marker primers, if the DNA of other white-fleshed Ganoderma lucidum varieties is detected by agarose gel electrophoresis, the PCR product will be negative, thereby achieving the successful, rapid, and accurate identification of the white-fleshed Ganoderma lucidum variety within the species. Application number: CN201710421417.9, invention name: A primer pair for identifying white-fleshed Ganoderma lucidum and its application. The present invention discloses a primer pair for identifying white-fleshed Ganoderma lucidum and its application. The primer pair and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5-6. (1) The primer pair of the invention has good specificity and can only amplify the specific band of white-fleshed Ganoderma lucidum; (2) The primer pair of the invention has high sensitivity and can quickly and accurately identify white-fleshed Ganoderma lucidum.

[0005] Xie Rong, et al., HPLC analysis and content determination of triterpenoids in Tibetan white meat Ganoderma lucidum, Tibet Agricultural Science and Technology, Vol. 43, No. 4, 2021, reported that the chromatographic conditions were: flow rate 1 mL / min, chromatographic column YMC, C 18 , 250mm×4.6mm, detection wavelength 254nm, mobile phase acetonitrile-acid water (pure water + 0.01% trifluoroacetic acid) gradient elution, the triterpene content of different forms of samples of white-fleshed Ganoderma lucidum cultivated in a solar greenhouse was analyzed; Wang Juan et al., Analysis of Volatile Components of Ganoderma lucidum Fruiting Bodies of Different Varieties, Chinese Edible Fungi, Vol. 40, No. 11, 2021, reported the use of gas chromatography to analyze the volatile components of the fruiting bodies of white-fleshed Ganoderma lucidum, red Ganoderma lucidum, purple Ganoderma lucidum, and big red Ganoderma lucidum.

[0006] The 2020 edition of the "Chinese Pharmacopoeia" states that Ganoderma lucidum comes from red Ganoderma lucidum and purple Ganoderma lucidum, while white Ganoderma lucidum contains triterpenes and polysaccharides that are not isolated in traditional varieties of Ganoderma lucidum. Currently, there is a lack of quality control indicators related to the properties and quality of white Ganoderma lucidum medicinal materials. This indicator is closely related to the composition of white Ganoderma lucidum and the identification of authentic medicinal materials, as well as the identification of artificially cultivated and wild white Ganoderma lucidum. However, the quality differences between authentic and fake white Ganoderma lucidum, and artificially cultivated and wild medicinal materials are still unclear; the reported chromatographic methods for white Ganoderma lucidum have not yet distinguished the authenticity of white Ganoderma lucidum. Summary of the Invention

[0007] The technical solution of the present invention provides a white-fleshed Ganoderma lucidum standard medicinal material extract and its detection method and use. The method can be used to identify the authenticity of white-fleshed Ganoderma lucidum medicinal materials and artificially cultivated and wild medicinal materials.

[0008] The invention provides a white-fleshed ganoderma lucidum standard medicinal material extract, which is an anhydrous ethanol extract of the white-fleshed ganoderma lucidum medicinal material Ganodermaleucocontextum THLi et al., wherein the HPLC fingerprint of the extract comprises 22 characteristic peaks, with peak 2 being used as a reference peak, and the relative retention times of the characteristic peaks are 0.210, 1.000, 1.075, 1.212, 1.463, 1.756, 2.504, 2.580, 2.741, 2.830, 3.065, 3.134, 3.213, 3.351, 3.494, 3.712, 4.163, 4.271, 4.519, 4.788, 4.840, and 4.903, respectively, and the relative retention times fluctuate within a range of ±10%.

[0009] The chromatographic conditions are:

[0010] Chromatographic column: Agilent Eclipse plus C 18 (4.6 mm × 250 mm, 5 μm), mobile phase: A acetonitrile-B 0.05% phosphoric acid water, detection wavelength: 254 nm; column temperature: 35°C; injection volume: 10 μL; flow rate: 1.0 mL min -1 ; Signal acquisition time: 110 min, gradient elution program: 0-50 min, 31%-45% A; 50-55 min, 45%-60% A; 55-100 min, 60%-90% A; 100-110 min, 90% A.

[0011] The average relative peak areas of the characteristic peaks are: 0.516, 1.000, 0.520, 1.734, 0.584, 0.754, 0.415, 1.333, 0.599, 0.403, 3.322, 0.576, 0.864, 0.570, 0.881, 0.474, 0.511, 0.459, 0.467, 0.468, 0.553, 0.603, and the average relative peak areas fluctuate within the range of ±10%.

[0012] Wherein, the preparation method of the extract is:

[0013] Weigh the white-fleshed Ganoderma lucidum medicinal material powder, add anhydrous ethanol, perform ultrasonic extraction, filter, wash the filter residue with 95% ethanol, evaporate the filtrate, make up the volume of the residue with 95% ethanol, filter through a microporous filter membrane, and obtain the white-fleshed Ganoderma lucidum medicinal material extract.

[0014] The present invention also provides the use of the white-fleshed Ganoderma lucidum standard medicinal material extract as a standard reference substance for detecting counterfeit white-fleshed Ganoderma lucidum medicinal materials, distinguishing artificial or wild medicinal materials from their sources, and medicinal material products.

[0015] The present invention provides a method for detecting a standard medicinal material of white-fleshed Ganoderma lucidum or an extract thereof, which comprises the following steps:

[0016] a. Preparation of standard: Take the white flesh Ganoderma lucidum standard medicinal material, add anhydrous ethanol, ultrasonically extract, filter, wash the filter residue with 95% ethanol, evaporate the filtrate to dryness, make up the volume with 95% ethanol, and filter through a microporous filter membrane to obtain the white flesh Ganoderma lucidum medicinal material extract to be tested;

[0017] b. HPLC detection was performed under the following chromatographic conditions: chromatographic column: Agilent Eclipse plus C 18 (4.6 mm × 250 mm, 5 μm), mobile phase: A acetonitrile-B 0.05% phosphoric acid water, detection wavelength: 254 nm; column temperature: 35°C; injection volume: 10 μL; flow rate: 1.0 mL min -1 ; Signal acquisition time: 110 min, gradient elution program: 0-50 min, 31%-45% A; 50-55 min, 45%-60% A; 55-100 min, 60%-90% A; 100-110 min, 90% A.

[0018] The present invention provides a method for identifying the authenticity of white-fleshed Ganoderma lucidum medicinal material, which comprises the following steps:

[0019] a. Using the white flesh Ganoderma lucidum standard medicinal material extract as a reference substance, the 22 common peak areas in the fingerprint were used as variables, SIMCA14.1 software was used for standardization, cluster analysis was performed on the data, a PCA model was established, and the equal variance method and UV method were used for scaling. Two principal components were extracted to obtain model explanatory parameters and predictive ability parameters; according to the principal component analysis results, the original variables of the two principal components were not less than 70%;

[0020] b. Take the medicinal material to be tested and obtain the fingerprint spectrum according to the above detection method;

[0021] c. Perform partial least squares discriminant analysis (PLS-DA) on the relative peak area results of the characteristic peaks of the sample to be tested and the relative peak area results of the characteristic peaks of the standard medicinal material extract in step a;

[0022] d. Analyze the results.

[0023] Further preferably, the model interpretation parameter R in step a is 2 X is 0.75, the prediction ability parameter Q 2 The PC1 score of the sample was 54.20%, the PC2 score was 20.40%, and the sum of the two was 74.60%.

[0024] The present invention provides a method for distinguishing artificially cultivated and wild white-fleshed Ganoderma lucidum, which comprises the following steps:

[0025] a. Take the medicinal material to be tested and obtain the fingerprint spectrum according to the above detection method;

[0026] b. Perform orthogonal partial least squares discriminant analysis (OPLS-DA) on the relative peak area results of the characteristic peaks of the sample to be tested and the relative peak area results of the characteristic peaks of the standard medicinal material extract in step a;

[0027] c. Analyze the results.

[0028] The present invention establishes a fingerprint of white-fleshed Ganoderma lucidum and performs chemometric analysis to achieve the authenticity identification of white-fleshed Ganoderma lucidum, as well as the identification of artificially cultivated and wild white-fleshed Ganoderma lucidum. The detection method of the present invention has strong repeatability, good precision, stability, and strong operability, and can identify the authenticity of white-fleshed Ganoderma lucidum medicinal materials, as well as the varieties of artificially cultivated and wild white-fleshed Ganoderma lucidum with high accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Fingerprints of 10 batches of white-fleshed Ganoderma lucidum

[0030] Figure 2 HCA analysis results of the authenticity of white Ganoderma lucidum

[0031] Figure 3 PCA analysis results

[0032] Figure 4 PLS-DA analysis results

[0033] Figure 5 HCA analysis results of cultivated and wild white-fleshed Ganoderma lucidum

[0034] Figure 6 OPLS-DA analysis results DETAILED DESCRIPTION

[0035] Example 1 Preparation method and detection method of the standard white flesh Ganoderma lucidum medicinal material extract of the present invention

[0036] 1. Source of medicinal materials

[0037] Ten batches of artificially cultivated white-fleshed Ganoderma lucidum (sample information is shown in Table 1) were identified as white-fleshed Ganoderma lucidum (Ganoderma leucocontextum) of the Polyporaceae family by Professor Zhao Bin of Guangdong Jiangmen Vocational College of Traditional Chinese Medicine.

[0038] Table 1 Sample information table

[0039]

[0040]

[0041] 2. Chromatographic conditions

[0042] Chromatographic column: Agilent Eclipse plus C 18 (4.6 mm × 250 mm, 5 μm), mobile phase: A acetonitrile-B 0.05% phosphoric acid water, detection wavelength: 254 nm; column temperature: 35°C; injection volume: 10 μL; flow rate: 1.0 mL min -1 ; Signal acquisition time: 110 min, gradient elution program: 0-50 min, 31%-45% A; 50-55 min, 45%-60% A; 55-100 min, 60%-90% A; 100-110 min, 90% A.

[0043] 3. Preparation of Test Solution

[0044] Weigh about 2 g of white-fleshed Ganoderma lucidum powder, accurately weigh it, accurately add 50 mL of anhydrous ethanol, weigh the mass, ultrasonically extract for 30 minutes, filter, wash the residue with 95% ethanol, evaporate the filtrate, dilute the residue to 2 ml with 95% ethanol, filter with a 0.45 μm microporous filter membrane, and set aside.

[0045] 4. Repeatability test

[0046] Accurately weigh 6 portions of S1 test sample, treat according to the method under "2", aspirate 10 μL, and inject according to the chromatographic conditions under "1". Use peak 2 as the reference peak to calculate the average relative retention time and average relative peak area RSD values of each common peak, which are 0.02% and 9.53%, respectively.

[0047] 5. Precision test

[0048] Accurately weigh the S1 test sample, process it according to the method under item "2", aspirate 10 μL, and continuously inject it 6 times according to the chromatographic conditions under item "1". Use peak 2 as the reference peak to calculate the average relative retention time and average relative peak area RSD values of each common peak, which are 0.04% and 4.76% respectively, indicating that the instrument has good precision.

[0049] 6. Stability test

[0050] Accurately weigh the S1 test sample, treat it according to the method under "2", aspirate 10 μL, and inject it at 0, 2, 4, 8, 12, and 24 h according to the chromatographic conditions under "1". Use peak 2 as the reference peak to calculate the average relative retention time and average relative peak area of each common peak. The RSD values are 0.04% and 6.18%, respectively, indicating that the test solution has good stability within 24 h.

[0051] 7. Establishment of fingerprint

[0052] The HPLC fingerprints of 10 batches of samples were analyzed using the Chinese medicine chromatographic fingerprint similarity evaluation system analysis software. After comparison, 22 common peaks were identified in the 10 batches of white meat Ganoderma lucidum samples. Figure 1 ,The similarity calculation results of the fingerprint maps are shown in Table 2 and Table 3.

[0053] Table 2 Similarity results of fingerprint spectra of 10 batches of white flesh Ganoderma lucidum

[0054]

[0055] Table 3 Average relative retention time and average relative peak area of common peaks

[0056]

[0057]

[0058] Example 2 Authenticity Identification Test of the White-fleshed Ganoderma Lucidum Medicinal Material of the Present Invention

[0059] 1. Source of medicinal materials

[0060] Ten batches of cultivated white Ganoderma were identified by Professor Zhao Bin of the Guangdong Jiangmen Vocational College of Traditional Chinese Medicine as Ganoderma leucocontextum, a Polyporaceae fungus. Their sources are listed in Table 1. Three other batches of counterfeit white Ganoderma were identified as Ganoderma lucidum (the fruiting body of the Polyporaceae fungus Ganoderma lucidum) and Ganoderma rubiginosa (the fruiting body of the Polyporaceae fungus Ganoderma leucocontextum). Sample information is listed in Table 4.

[0061] Table 4 Authenticity identification test sample information

[0062]

[0063] 2. Chemometric Analysis

[0064] 2.1 Cluster Analysis (HCA)

[0065] The 22 common peak areas obtained from the fingerprint were used as variables, and the data were standardized using SIMCA14.1 software. The clustering method was Ward, and cluster analysis was performed on the data. The results are shown in Figure 2 . 10 batches of white-fleshed Ganoderma lucidum and 3 batches of fake Ganoderma lucidum can be clustered into 2 categories, 10 batches of white-fleshed Ganoderma lucidum can be clustered into one category, and 3 batches of fake Ganoderma lucidum can be clustered into one category. It is possible to clearly distinguish between genuine and fake white-fleshed Ganoderma lucidum.

[0066] 2.2 Principal Component Analysis (PCA)

[0067] The 22 common peak areas of 13 batches of samples were normalized and imported into SIMCA14.1 software to establish a PCA model. The equal variance (UV) method was used for scaling and two principal components were extracted to obtain the model explanatory parameter R. 2 X is 0.75, and the prediction ability parameter Q2 is 0.52. The PCA score graph is as follows Figure 3 According to the principal component analysis, the sample has an important discriminant variable PC1 score of 54.20% and PC2 score of 20.40%, and the sum of the two is 74.60%, indicating that the two principal components extracted can explain 74.60% of the original variables and can explain most of the sample information.

[0068] 2.3 Partial Least Squares Discriminant Analysis (PLS-DA)

[0069] The common peak areas of the 13 batches of samples were normalized to form a 22×13 matrix and imported into SIMCA14.1 software for PLS-DA analysis. The results are shown in Figure 4 The fitting parameters of the established PLS-DA model, model parameter R 2 X is 0.70, R 2 Y is 0.91, which is greater than 0.5, indicating that the model has good stability and predictive ability. Figure 4 It can be seen that compared with the PCA model, the samples can be clearly divided into two categories, the classification effect is significant, and the genuine and counterfeit white-fleshed Ganoderma lucidum can be clearly distinguished, which is consistent with the clustering results, indicating that this method has a better classification effect.

[0070] Example 3 Method for Identifying Cultivated and Wild White Ganoderma Lucidum

[0071] Ten batches of cultivated white-fleshed Ganoderma lucidum were identified as Ganoderma leucocontextum, a Polyporaceae fungus, by Professor Zhao Bin of the Guangdong Jiangmen Vocational College of Traditional Chinese Medicine. Details are shown in Table 1. Two batches of wild white-fleshed Ganoderma lucidum were also found; sample information is shown in Table 5.

[0072] Table 5 Sample information of artificial cultivation and wild identification test

[0073]

[0074] 2. Cluster Analysis (HCA)

[0075] The 22 common peak areas obtained from the fingerprint were used as variables, and the data were standardized using SIMCA14.1 software. The clustering method was Ward, and cluster analysis was performed on the data. The results are shown in Figure 5. 10 batches of artificially cultivated white-fleshed Ganoderma lucidum and 2 batches of wild white-fleshed Ganoderma lucidum can be clustered into 2 categories, 10 batches of artificially cultivated white-fleshed Ganoderma lucidum can be clustered into one category, and 2 batches of wild white-fleshed Ganoderma lucidum can be clustered into one category. It is possible to clearly distinguish artificially cultivated and wild white-fleshed Ganoderma lucidum.

[0076] 3. Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA)

[0077] The common peak areas of the 13 batches of samples were normalized to form a 22×13 matrix and imported into SIMCA14.1 software for OPLS-DA analysis. The results are shown in Figure 6 .from Figure 6 It can be seen that the samples can be clearly divided into two categories, and the classification effect is significant. It can clearly distinguish between wild and artificially cultivated white-fleshed Ganoderma lucidum, which is consistent with the results of cluster analysis, indicating that this method has a good classification effect.

[0078] Example 3 Verification experiment of the present invention

[0079] The method of the present invention was used to purchase Ganoderma lucidum medicinal materials from different sources, and a verification test of artificial cultivation and wild white Ganoderma lucidum identification and authenticity verification test was conducted. The results showed that the accuracy of the method was higher than 80%. The results are shown in Tables 6 and 7.

[0080] Table 6 Authenticity verification test results

[0081]

[0082] Table 7 Results of verification test on artificial cultivation and wild identification

[0083]

[0084] In summary, the method for analyzing the results of the present invention is to identify the common peaks by detecting the fingerprint of white-fleshed Ganoderma lucidum, and to clarify the correlation between the peak area and the quality of different varieties of medicinal materials by analyzing the average relative peak area of the common peaks. The quality of white-fleshed Ganoderma lucidum medicinal materials is comprehensively evaluated from multiple indicators, and the method is easy to operate and can be used to identify the authenticity of white-fleshed Ganoderma lucidum.

Claims

1. A method for constructing a fingerprint of a standard medicinal material of white flesh Ganoderma lucidum or its extract, characterized by: It includes the following steps: a. Preparation of standard: Take the white flesh Ganoderma lucidum standard medicinal material, add anhydrous ethanol, ultrasonically extract, filter, wash the filter residue with 95% ethanol, evaporate the filtrate to dryness, dilute the residue with 95% ethanol, and filter through a microporous filter membrane to obtain the white flesh Ganoderma lucidum medicinal material extract to be tested; b. HPLC detection, chromatographic conditions are: chromatographic column: Agilent Eclipse plus C 18 , specifications: 4.6mm×250mm, 5μm, mobile phase: A acetonitrile-B 0.05% phosphoric acid water, detection wavelength: 254 nm; column temperature: 35℃; injection volume: 10 μL; flow rate: 1.0 mL·min -1 ; Signal acquisition time: 110 min, gradient elution program: 0-50 min, 31%-45%A; 50-55 min, 45%-60%A; 55-100 min, 60%-90% A; 100-110 min, 90%A.

2. The method for constructing the fingerprint of the standard medicinal material of white flesh Ganoderma lucidum or its extract according to claim 1, characterized in that: The HPLC fingerprint contains 22 characteristic peaks, with peak 2 as the reference peak. The relative retention times of the characteristic peaks are 0.210, 1.000, 1.075, 1.212, 1.463, 1.756, 2.504, 2.580, 2.741, 2.830, 3.065, 3.134, 3.213, 3.351, 3.494, 3.712, 4.163, 4.271, 4.519, 4.788, 4.840, and 4.903, and the relative retention times fluctuate within the range of ±10%.

3. The method for constructing the fingerprint of the standard medicinal material of white flesh Ganoderma lucidum or its extract according to claim 2, characterized in that: The average relative peak areas of the characteristic peaks are: 0.516, 1.000, 0.520, 1.734, 0.584, 0.754, 0.415, 1.333, 0.599, 0.403, 3.322, 0.576, 0.864, 0.570, 0.881, 0.474, 0.511, 0.459, 0.467, 0.468, 0.553, 0.603, and the average relative peak areas fluctuate within the range of ±10%.

4. Use of the method for constructing a fingerprint of a standard medicinal material of white-fleshed Ganoderma lucidum or an extract thereof according to any one of claims 1 to 3 in detecting counterfeit white-fleshed Ganoderma lucidum medicinal materials, distinguishing between artificial and wild medicinal materials, and standard reference substances for medicinal material products.

5. A method for identifying the authenticity of white-fleshed Ganoderma lucidum medicinal material, characterized by: It includes the following steps: a. The standard medicinal material extract of white flesh Ganoderma lucidum was used as the reference substance. The peak areas of 22 common peaks in the fingerprint were used as variables. SIMCA14.1 software was used for standardization processing. Cluster analysis was performed on the data. A PCA model was established. The equal variance method and UV method were used for scaling. Two principal components were extracted to obtain the model explanatory parameters and predictive ability parameters. According to the principal component analysis results, the original variables of the two principal components were not less than 70%; b. Take the medicinal material to be tested and obtain a fingerprint according to the construction method of claim 1; c. Perform partial least squares discriminant analysis (PLS-DA) on the relative peak area results of the characteristic peaks of the samples to be tested and the relative peak area results of the characteristic peaks of the standard medicinal material extracts in step a; d. Analyze the results.

6. The method for identifying the authenticity of white-fleshed Ganoderma lucidum medicinal material according to claim 5, characterized in that: The model explanation parameter R in step a is 2 X The prediction ability parameter Q is 0.

73. 2 The PC1 score of the sample was 57.80%, and the PC2 score was 15.30%, respectively. The sum of the two was 73.10%.

7. A method for distinguishing cultivated and wild white-fleshed Ganoderma lucidum, characterized in that: It includes the following steps: a. Take the medicinal material to be tested and obtain a fingerprint according to the construction method of claim 1; b. Perform orthogonal partial least squares discriminant analysis (OPLS-DA) on the relative peak area results of the characteristic peaks of the sample to be tested and the relative peak area results of the characteristic peaks of the standard medicinal material extract in step a; c. Analyze the results.

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