A method for identifying tangerine peel from different raw material sources and its application

Abstract

The present invention provides a method for identifying tangerine peel from different raw material sources and its application. The identification method comprises the following steps: (1) mixing a sample to be tested with a first solvent for extraction, collecting the filtrate and then mixing it with a second solvent for extraction to obtain a test solution; (2) performing liquid chromatography detection on the test solution, and judging the raw material source of the sample to be tested by combining the detection result with a characteristic chromatogram. The identification method provided by the present invention can effectively and accurately identify and analyze the raw material source of tangerine peel, provides a new method for identifying tangerine peel from different raw material sources, and greatly expands the application prospect of tangerine peel.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine analysis and identification, and specifically relates to a method for identifying tangerine peels from different raw material sources and an application thereof, and in particular relates to an accurate method for identifying tangerine peels from different raw material sources and an application thereof. Background Art

[0002] At present, the identification of Chinese herbal medicines under the Chinese Pharmacopoeia mainly includes property identification, microscopic identification, and physical and chemical identification. Among them, physical and chemical identification includes physical identification, chemical identification, spectral identification, chromatographic identification, etc. Chromatographic identification is still mainly based on thin-layer chromatography identification. However, the identification of different varieties of Chinese herbal medicines is rarely reflected in the Chinese Pharmacopoeia. The main reason is that there is a lack of research on the differences in the material components of different varieties of Chinese herbal medicines, and the differences in chemical information are difficult to control.

[0003] CN109374762B discloses a method for identifying Guang Chenpi and Chenpi varieties based on metabolomics. It includes: preparing Guang Chenpi and Chenpi samples, detecting by ultra-high performance liquid phase-mass spectrometry, obtaining LC-MS data, and identifying compounds in Guang Chenpi and Chenpi samples using UNIFI and online biological database. Then, after pre-processing the LC-MS raw data of Guang Chenpi and Chenpi, SIMCA-P14.0 software is imported for multivariate statistical analysis to obtain the marker metabolites of Guang Chenpi and Chenpi, and the change trend of the marker metabolites is intuitively displayed through a heatmap. The analysis results of the invention are displayed in the form of identified compounds, PCA score graphs, OPLS-DA score graphs, s-plot graphs, and heatmap graphs, clearly revealing the differences between Guang Chenpi and Chenpi, indicating that metabolomics can be used to distinguish Guang Chenpi and Chenpi, and the technical methods are advanced and the experimental results are reliable, providing an excellent method for the identification and quality evaluation of Chenpi varieties.

[0004] CN109142590B discloses a method and device for identifying and classifying Guang Chenpi, which belongs to the technical field of identification of traditional Chinese medicine. The identification and classification method comprises the following steps: establishing a standard Guang Chenpi database: taking a Guang Chenpi sample, using gas chromatography ion mobility spectrometry to detect, obtaining GC-IMS analysis data of standard Guang Chenpi, and establishing a standard Guang Chenpi database; identification and determination: using a gas chromatography ion mobility spectrometry instrument to detect the tangerine peel sample to be tested according to the detection method of the Guang Chenpi sample, obtaining GC-IMS analysis data of the tangerine peel sample to be tested, extracting characteristic data therein, analyzing and comparing with the data in the standard Guang Chenpi database, and determining the type of the tangerine peel sample to be tested. The method has good repeatability, is simple and fast, and has a wide range of application value.

[0005] In the pharmacopoeia, only tangerine peel and Guangdong tangerine peel are included in the tangerine peel medicinal material, and the main cultivated varieties are tea-branch tangerine peel (Guangzhou tangerine peel), Dahongpao, Wenzhou mandarin orange, and Fuju, but there is a lack of specific identification methods between cultivated varieties. At present, the price of Guangdong tangerine peel medicinal materials circulating on the market is more than 20 times higher than that of general tangerine peel (the price is 6 to 8 yuan / kg), and there are also certain differences in their pharmacologically active ingredients. Therefore, establishing a specific identification method between different cultivated varieties of tangerine peel medicinal materials will have a positive impact on the current research of new Chinese medicines, especially classic prescriptions, in order to fix the factors that affect the stability of medicinal materials, such as the origin, origin, processing method, harvesting period, etc., to ensure the stability of the quality of new Chinese medicines, especially classic prescriptions. Although there are articles that have studied the fingerprints of active ingredients of different tangerine peels, there are no reports on the specific identification methods of tangerine peel (red tangerine, ponkan) and tangerine peel (mandarin orange), which greatly limits the reasonable application of tangerine peel medicinal materials of different cultivated varieties. Therefore, how to provide an accurate identification method of dried tangerine peel (red orange, ponkan) and dried tangerine peel (honey orange) has become a problem to be solved urgently. Summary of the invention

[0006] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a method for identifying tangerine peel from different raw material sources and its application, and in particular to provide an accurate method for identifying tangerine peel from different raw material sources and its application. The identification method provided by the present invention can effectively and accurately identify and analyze the raw material sources of tangerine peel, provides a new method for identifying tangerine peel from different raw material sources, and greatly expands the application prospects of tangerine peel.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In one aspect, the present invention provides a method for identifying tangerine peel from different raw material sources, the identification method comprising the following steps:

[0009] (1) extracting the sample to be tested by mixing it with a first solvent, collecting the filtrate and then extracting it with a second solvent to obtain a test solution;

[0010] (2) The liquid to be tested is subjected to liquid chromatography detection, and the detection result is combined with the characteristic spectrum to determine the source of the raw materials of the sample to be tested.

[0011] The identification method is used to identify tangerine peel made from red mandarin orange and tangerine peel made from mandarin orange, or to identify tangerine peel made from ponkan orange and tangerine peel made from mandarin orange.

[0012] In step (1), the first solvent comprises water.

[0013] In step (1), the second solvent comprises methanol.

[0014] The above method can effectively distinguish the raw material sources of the samples to be tested with high accuracy by effectively extracting the samples to be tested, then performing liquid-phase detection, and combining specific characteristic spectra.

[0015] Preferably, the material-liquid ratio of the sample to be tested to the first solvent in step (1) is 1:(5 - 15).

[0016] Preferably, the temperature for mixing and extracting with the first solvent in step (1) is 96 - 100 °C, and the time is 30 - 60 min.

[0017] Preferably, the volume ratio of the second solvent to the filtrate in step (1) is (2.5 - 3.5):1.

[0018] Preferably, the temperature for mixing and extracting with the second solvent in step (1) is 10 - 35 °C, and the time is 5 - 15 min.

[0019] Among them, the material-liquid ratio of the sample to be tested to the first solvent can be 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14 or 1:15, etc.; the temperature for mixing and extracting with the first solvent can be 96 °C, 97 °C, 98 °C, 99 °C or 100 °C, etc.; the time can be 30 min, 35 min, 40 min, 45 min, 50 min, 55 min or 60 min, etc.; the volume ratio of the second solvent to the filtrate can be 2.5:1, 2.6:1, 2.7:1, 2.8:1, 2.9:1, 3:1, 3.1:1, 3.2:1, 3.3:1, 3.4:1 or 3.5:1, etc.; the temperature for mixing and extracting with the second solvent can be 10 °C, 15 °C, 20 °C, 25 °C, 30 °C or 35 °C, etc.; the time can be 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, 11 min, 12 min, 13 min, 14 min or 15 min, etc. However, it is not limited to the above-listed values, and other unlisted values within the above numerical range are equally applicable.

[0020] Preferably, in the liquid chromatography detection of step (2), the flow rate of the mobile phase is 0.8 - 1.2 mL / min.

[0021] Preferably, the mobile phase includes mobile phase A and mobile phase B. Mobile phase A is acetonitrile, mobile phase B is phosphoric acid solution, and the volume fraction of phosphoric acid in the phosphoric acid solution is 0.08 - 0.12%.

[0022] Preferably, the wavelength for the liquid chromatography detection is 310 - 330 nm.

[0023] Among them, the flow rate of the mobile phase can be 0.8 mL / min, 0.9 mL / min, 1 mL / min, 1.1 mL / min, 1.2 mL / min, etc., the volume fraction of phosphoric acid can be 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, etc., and the wavelength can be 310 nm, 315 nm, 320 nm, 325 nm, 330 nm, etc., but not limited to the values listed above. Other unlisted values within the above numerical ranges are equally applicable.

[0024] Preferably, the elution process of the liquid chromatography detection in step (2) is as follows:

[0025] At 0 - 45 min, the volume fraction of mobile phase A changes uniformly from 4.5 - 5.5% to 15.5 - 16.5%, and the volume fraction of mobile phase B changes uniformly from 94.5 - 95.5% to 83.5 - 84.5%;

[0026] At 45 - 65 min, the volume fraction of mobile phase A changes uniformly from 15.5 - 16.5% to 23.5 - 24.5%, and the volume fraction of mobile phase B changes uniformly from 83.5 - 84.5% to 75.5 - 76.5%;

[0027] At 65 - 90 min, the volume fraction of mobile phase A changes uniformly from 23.5 - 24.5% to 59.5 - 60.5%, and the volume fraction of mobile phase B changes uniformly from 75.5 - 76.5% to 39.5 - 40.5%;

[0028] At 90 - 91 min, the volume fraction of mobile phase A changes uniformly from 59.5 - 60.5% to 4.8 - 5.2%, and the volume fraction of mobile phase B changes uniformly from 39.5 - 40.5% to 94.8 - 95.2%.

[0029] Among them, from 0 to 45 minutes, the initial volume fraction of mobile phase A can be 4.5%, 4.7%, 4.9%, 5.1%, 5.3% or 5.5%, etc., and the initial volume fraction of mobile phase B can be 94.5%, 94.7%, 94.9%, 95.1%, 95.3% or 95.5%, etc. The volume fraction of mobile phase A can change to 15.5%, 15.7%, 15.9%, 16.1%, 16.3 or 16.5%, etc., and the volume fraction of mobile phase B can change to 83.5%, 83.7%, 83.9%, 84.1%, 84.3% or 84.5%, etc. From 45 to 65 minutes, the volume fraction of mobile phase A can change to 23.5%, 23.7%, 23.9%, 24.1%, 24.3% or 24.5%, etc., and the volume fraction of mobile phase B can change to 75.5%, 75.7%, 75.9%, 76.1%, 76.3% or 76.5%, etc. From 65 to 90 minutes, the volume fraction of mobile phase A can change to 59.5%, 59.7%, 59.9%, 60.1%, 60.3% or 60.5%, etc., and the volume fraction of mobile phase B can change to 39.5%, 39.7%, 39.9%, 40.1%, 40.3% or 40.5%, etc. From 90 to 91 minutes, the change value of the volume fraction of mobile phase A can be 4.8%, 4.9%, 5.0%, 5.1% or 5.2%, etc., and the volume fraction of mobile phase B can change to 94.8%, 94.9%, 95.0%, 95.1% or 95.2%, etc., but not limited to the values listed above. Other unlisted values within the above numerical range are equally applicable.

[0030] The above control of specific detection parameters can effectively improve the accuracy of detection.

[0031] Preferably, the characteristic chromatogram in step (2) is obtained by a method including the following steps:

[0032] (1’) Treat the dried tangerine peels made from red tangerines or ponkan oranges and the dried tangerine peels made from satsumas respectively according to the above-mentioned extraction and detection methods to obtain liquid phase results;

[0033] (2’) Compare the liquid phase results of the dried tangerine peels made from red tangerines or ponkan oranges and the dried tangerine peels made from satsumas. Take the hesperidin peak as the S peak, and take D-glucaric acid ((E,E)-2,4-bis[3-(4-hydroxy-3-methoxyphenyl)-2-propenoate] D-glucaric acid) and auraptene as the characteristic peaks of the dried tangerine peels made from red tangerines or ponkan oranges, and take naringin, apamin and 3,5,6,7,8,3',4'-heptamethoxyflavone as the characteristic peaks of the dried tangerine peels made from satsumas, and calculate the relative retention time of each characteristic peak relative to the S peak.

[0034] The tangerine peel made from red tangerine or ponkan and the tangerine peel made from satsuma mandarin for preparing the characteristic chromatogram are of known origin and can be obtained on the market.

[0035] Preferably, the relative retention time of D-glucaric acid ((E,E)-2,4-bis[3-(4-hydroxy-3-methoxyphenyl)-2-propenoate] D-glucarate) is 0.84 - 0.93, such as 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92 or 0.93, etc.; the relative retention time of auraptene is 1.25 - 1.39, such as 1.25, 1.26, 1.27, 1.28, 1.29, 1.30, 1.31, 1.32, 1.33, 1.34, 1.35, 1.36, 1.37, 1.38 or 1.39, etc., but not limited to the values listed above, and other unlisted values within the above numerical range are equally applicable.

[0036] Preferably, the relative retention time of naringin is 0.88 - 0.98, the relative retention time of apamin is 1.16 - 1.28, and the relative retention time of 3,5,6,7,8,3',4'-heptamethoxyflavone is 1.32 - 1.46.

[0037] Preferably, the specific process of combining the test result with the characteristic chromatogram to judge the raw material source of the sample to be tested is as follows: compare the test result with the characteristic chromatogram, observe whether characteristic peaks appear in the test result, calculate the relative retention time of the characteristic peaks, and then compare with the characteristic chromatogram to judge the coincidence situation, so as to judge that the sample to be tested is tangerine peel made from red tangerine or ponkan or tangerine peel made from satsuma mandarin.

[0038] On the other hand, the present invention also provides the application of the identification method as described above in the identification of traditional Chinese medicines.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] The present invention provides a method for identifying tangerine peel from different raw material sources. By effectively extracting the sample to be tested, then performing liquid phase detection, and combining with a specific characteristic chromatogram, the raw material source of the sample to be tested can be effectively distinguished with high accuracy. Description of the Drawings

[0041] Figure 1 It is the liquid phase result diagram of the tangerine peel medicinal material in the process of preparing the characteristic chromatogram in Example 1;

[0042] Figure 2 It is the liquid phase result diagram of the hesperidin reference substance in the process of preparing the characteristic chromatogram in Example 1. Detailed Embodiments

[0043] The technical solution of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations on the present invention.

[0044] In the following examples, the instrument used was a Shimadzu high-performance liquid chromatography integrated instrument (LC-2030C 3D Plus); the chromatographic column was Waters XSelect HSS T3 (5μm, 4.6×250mm).

[0045] Example 1

[0046] This example provides a method for identifying tangerine peels from different raw material sources. The specific steps are as follows:

[0047] Preparation of characteristic chromatogram:

[0048] Take 10 g of standard tangerine peel herbs (sieved through No. 1 sieve, sourced from two different batches of red oranges, one batch of ponkan oranges, and three different batches of satsumas), place them in a round-bottom flask, add 10 times the amount of water (100 mL), soak for 30 min, heat to boiling and maintain gentle boiling for 45 min, filter while it is hot, and after cooling to 20°C, take 1 mL of the water extract, add 3 mL of methanol, shake well, centrifuge, take the supernatant, and filter to obtain the test solution. Perform liquid chromatography detection on the test solution, use acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, and perform gradient elution according to the regulations in the following table; the flow rate is 1.0 mL per minute; the column temperature is 30°C; the detection wavelength is 320 nm, and the injection volume is 10 μL.

[0049]

[0050] The test results are as Figure 1 shown. The main peaks are marked with numbers. In addition, the hesperidin reference substance was detected according to the above method, and the results are as Figure 2 shown. It can be found from the figure that peak 8 is the hesperidin peak, with strong absorption, moderate retention time, and baseline separation, so it is selected as the S peak.

[0051] In addition, peaks 7 and 9 in tangerine peels (red oranges, ponkan oranges) only appear in the test results of tangerine peels (red oranges, ponkan oranges), and the characteristic peaks in tangerine peels (red oranges) and tangerine peels (ponkan oranges) are the same; peaks 7, 9, and 11 in tangerine peels (satsumas) only appear in the test results of tangerine peels (satsumas). Therefore, peaks 7 and 9 are selected as the characteristic peaks of tangerine peels (red oranges, ponkan oranges), and peaks 7, 9, and 11 are selected as the characteristic peaks of tangerine peels (satsumas), and their mass spectrometry characterizations are as follows:

[0052]

[0053] Afterwards, calculate the relative retention times of each characteristic peak. The relative retention time of D-glucaric acid ((E,E)-2,4-bis[3-(4-hydroxy-3-methoxyphenyl)-2-propenoate] D-glucarate) is 0.89, that of auraptene is 1.32, that of naringin is 0.93, that of melittin is 1.22, and that of 3,5,6,7,8,3',4'-heptamethoxyflavone is 1.39.

[0054] Respectively weigh 10 g of the medicinal material powders (passing through No. 1 sieve) of the samples to be tested, namely ponkan oranges (batch number: YC2020010503), ponkan oranges (batch number: YC2019110406), red tangerines (batch number: YC2019110201), satsumas (batch number: YC2019110501), satsumas (batch number: YC2019110402), and satsumas (batch number: YC2019110301), place them in a round-bottom flask, add 10 times the amount of water (100 mL), soak for 30 min, heat to boiling and maintain gentle boiling for 45 min, filter while it is hot, and after cooling to 20 °C, take 1 mL of the water extract, add 3 mL of methanol, shake well, centrifuge, take the supernatant, and filter to obtain the test solution. Perform liquid chromatography detection on the test solution, using acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, and perform gradient elution according to the regulations in the following table; the flow rate is 1.0 mL per minute; the column temperature is 30 °C; the detection wavelength is 320 nm, and the injection volume is 10 μL.

[0055]

[0056]

[0057] Compare the test results with the characteristic chromatogram. The results show that the relative retention time of D-glucaric acid ((E,E)-2,4-bis[3-(4-hydroxy-3-methoxyphenyl)-2-propenoate] D-glucarate) is 0.89 and that of auraptene is 1.31, which are consistent with the characteristic chromatograms of red tangerines and ponkan oranges; the relative retention time of naringin is 0.94, that of melittin is 1.21, and that of 3,5,6,7,8,3',4'-heptamethoxyflavone is 1.38, which are consistent with the characteristic chromatogram of satsumas, and effective identification has been achieved for all.

[0058] Precision:

[0059] Perform a precision test on the method provided in Example 1, and the results are as follows:

[0060] Table 1 Precision test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - red tangerine and ponkan orange varieties

[0061]

[0062] Table 2 Precision test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - satsuma variety

[0063]

[0064]

[0065] Repeatability:

[0066] The repeatability test was carried out on the method provided in Example 1, and the results are as follows:

[0067] Table 3 Repeatability test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - Red tangerine and Ponkan varieties

[0068]

[0069] Table 4 Repeatability test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - Mandarin orange variety

[0070]

[0071]

[0072] Stability:

[0073] The stability test was carried out on the method provided in Example 1, and the results are as follows:

[0074] Table 5 Stability test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - Red tangerine and Ponkan varieties

[0075]

[0076] Table 6 Stability test - RSD (%) results of relative retention time and relative peak area of characteristic peaks - Mandarin orange variety

[0077]

[0078]

[0079] The above results show that the identification method provided by the present invention has high precision, good repeatability and good stability, and has excellent effects.

[0080] The applicant declares that the present invention illustrates the identification method and its application of tangerine peel from different raw material sources through the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

[0081] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0082] In addition, it should be noted that, in the above specific embodiments, the various specific technical features described can be combined in any suitable manner without conflict. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A method for identifying tangerine peel from different raw material sources, characterized in that, The identification method includes the following steps: (1) Mix the sample to be tested with water for extraction, collect the filtrate and then mix it with methanol for extraction to obtain the test solution; (2) Perform liquid chromatography detection on the test solution. The liquid chromatography detection conditions are as follows: the flow rate of the mobile phase is 0.8 - 1.2 mL / min. The mobile phase includes mobile phase A and mobile phase B. Mobile phase A is acetonitrile, and mobile phase B is a phosphoric acid solution. The volume fraction of phosphoric acid in the phosphoric acid solution is 0.08 - 0.12%. The detection wavelength is 310 - 330 nm. The chromatographic column is Waters XSelect HSS T3, with a specification of 5 μm, 4.6×250 mm. The elution process of the liquid chromatography detection is as follows: From 0 to 45 min, the volume fraction of mobile phase A changes uniformly from 4.5 - 5.5% to 15.5 - 16.5%, and the volume fraction of mobile phase B changes uniformly from 94.5 - 95.5% to 83.5 - 84.5%; From 45 to 65 min, the volume fraction of mobile phase A changes uniformly from 15.5 - 16.5% to 23.5 - 24.5%, and the volume fraction of mobile phase B changes uniformly from 83.5 - 84.5% to 75.5 - 76.5%; From 65 to 90 min, the volume fraction of mobile phase A changes uniformly from 23.5 - 24.5% to 59.5 - 60.5%, and the volume fraction of mobile phase B changes uniformly from 75.5 - 76.5% to 39.5 - 40.5%; From 90 to 91 min, the volume fraction of mobile phase A changes uniformly from 59.5 - 60.5% to 4.8 - 5.2%, and the volume fraction of mobile phase B changes uniformly from 39.5 - 40.5% to 94.8 - 95.2%; (3) Combine the detection results with the characteristic chromatogram to judge the raw material source of the sample to be tested; The identification method is used to identify tangerine peel made from red tangerines and tangerine peel made from satsumas, or to identify tangerine peel made from ponkan oranges and tangerine peel made from satsumas; The characteristic chromatogram in step (3) is obtained by a method including the following steps: (1') Process the tangerine peel made from red tangerines or ponkan oranges and the tangerine peel made from satsumas respectively according to the extraction and detection methods described in steps (1) - (2) to obtain the liquid phase results; (2') Compare the liquid phase results of the tangerine peel made from red tangerines or ponkan oranges and the tangerine peel made from satsumas. Take the hesperidin peak as the S peak, and take D - glucaric acid ((E,E)-2,4 - bis[3-(4 - hydroxy - 3 - methoxyphenyl)-2 - acrylate] D - glucarate), auraptene as the characteristic peaks of the tangerine peel made from red tangerines or ponkan oranges, and take naringin, apamin, 3,5,6,7,8,3',4'-heptamethoxyflavone as the characteristic peaks of the tangerine peel made from satsumas, and calculate the relative retention time of each characteristic peak relative to the S peak.

2. The identification method according to claim 1, characterized in that, The material - liquid ratio of the sample to be tested and water in step (1) is 1:(5 - 15).

3. The identification method according to claim 1, characterized in that, The temperature of the extraction by mixing with water in step (1) is 96 - 100 °C, and the time is 30 - 60 min.

4. The identification method according to claim 1, characterized in that, The volume ratio of methanol to the filtrate in step (1) is (2.5 - 3.5):

1.

5. The identification method according to claim 1, characterized in that, The temperature for the mixed extraction with methanol in step (1) is 10 - 35 °C, and the time is 5 - 15 min.

6. The identification method according to claim 1, characterized in that, The relative retention time of D-glucaric acid ((E,E)-2,4-bis[3-(4-hydroxy-3-methoxyphenyl)-2-propenoate] D-gluconic acid) is 0.84 - 0.93, and the relative retention time of auraptene is 1.25 - 1.

39.

7. The identification method according to claim 1, characterized in that, The relative retention time of naringin is 0.88 - 0.98, the relative retention time of apamin is 1.16 - 1.28, and the relative retention time of 3,5,6,7,8,3',4'-heptamethoxyflavone is 1.32 - 1.

46.

8. The identification method according to claim 1, characterized in that, The specific process of judging the raw material source of the sample to be tested by combining the detection result with the characteristic chromatogram is as follows: compare the detection result with the characteristic chromatogram, observe whether characteristic peaks appear in the detection result, calculate the relative retention time of the characteristic peaks, and then compare with the characteristic chromatogram to judge the coincidence situation, so as to judge that the sample to be tested is tangerine peel made from red tangerine or ponkan or tangerine peel made from satsuma mandarin.

9. An application of the identification method according to any one of claims 1-8 in the identification of traditional Chinese medicines.

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