Identification method of Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. and its application

Abstract

The present invention provides a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. and its application. The identification method includes the following steps: (1) Mix the sample to be tested with a solvent for reflux extraction, collect the filtrate, mix it with an organic solution and let it stand still. Then take the supernatant for desolvation, dissolve the residue with an organic solution and filter it to obtain the test solution A; (2) Perform liquid chromatography detection on the test solution A obtained in step (1), compare the detection result with the characteristic chromatogram, and judge whether the sample to be tested is Euryale ferox Salisb. or stir-fried Euryale ferox Salisb. The identification method provided by the present invention for Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. has high accuracy and strong characteristics, and is of great significance for the quality control of clinical Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. decoction pieces.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine analysis, and particularly relates to a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. and its application, especially an accurate and highly characteristic method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. and its application. Background Art

[0002] Euryale ferox Salisb. is the dried ripe semen of the plant Euryale ferox Salisb. of the Nymphaeaceae family. It has a slight odor and a bland taste, and enters the spleen and kidney meridians. The raw Euryale ferox Salisb. has the effects of tonifying the kidney and securing essence, invigorating the spleen and stopping diarrhea, and eliminating dampness and arresting leukorrhea, and is used for spermatorrhea and emission, enuresis and frequent urination, chronic diarrhea due to spleen deficiency, leukorrhea, and vaginal discharge; Stir-fried Euryale ferox Salisb. is a processed product of Euryale ferox Salisb. decoction pieces, which has not been included in the current pharmacopoeia, and has the effects of invigorating the spleen and tonifying the kidney, and securing and arresting leukorrhea. After stir-frying and roasting, Euryale ferox Salisb. is warm in nature and has a slight fragrance, increasing its astringent property, and enhancing its effects of invigorating the spleen and tonifying the kidney, and securing and arresting leukorrhea.

[0003] At present, the identification of traditional Chinese medicine decoction pieces under the Chinese Pharmacopoeia mainly includes morphological identification, microscopic identification, physicochemical identification. Among them, physicochemical identification includes physical identification, chemical identification, spectral identification, chromatographic identification, etc., and chromatographic identification still mainly relies on thin-layer chromatographic identification. However, the specific identification of traditional Chinese medicinal materials and their processed products is rarely reflected in the Chinese Pharmacopoeia. The main reason is that there is insufficient research on the differences in the material components of traditional Chinese medicinal materials and their processed products, and it is difficult to control the chemical information differences.

[0004] Stir-fried Euryale ferox Salisb. is superior in invigorating the spleen and astringing, and is commonly used for diarrhea due to spleen deficiency and spermatorrhea caused by insufficient closure of the kidney essence, and can also be used for leukorrhea due to spleen deficiency, such as the classic famous prescription Yihuang Decoction (Fu Qingzhu's Gynecology), Ganshu Decoction recorded in Luo's Huiyue Yijing, and Suoyang Gujing Pills included in the Chinese Pharmacopoeia (2020 Edition). However, currently, only Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. with bran are included in the pharmacopoeia. The quality control only includes morphological identification, microscopic identification, thin-layer chromatographic identification and other inspection items. There is no identification method for the components of Euryale ferox Salisb., and stir-fried Euryale ferox Salisb. decoction pieces have not been included. Currently, it is difficult to control the quality of Euryale ferox Salisb. and its decoction pieces well. The identification of Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. can only rely on subjective morphological differences, which greatly limits the safe and reasonable application of Euryale ferox Salisb.

[0005] Su Guiyun et al. summarized the differences in appearance and odor between stir-fried Euryale ferox Salisb. with bran and Euryale ferox Salisb., but it is inevitable to have problems of misidentification when only examining from this aspect (Su Guiyun, Liu Guotong. Euryale ferox Salisb. and Stir-fried Euryale ferox Salisb. with Bran [J]. Capital Medicine, 2011, 000(015):49).

[0006] Since the current poor identification effect of Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. severely restricts the safe and reasonable application of Euryale ferox Salisb. Therefore, how to provide an accurate method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. has become an urgent problem to be solved. Summary of the Invention

[0007] In view of the deficiencies of the prior art, the purpose of the present invention is to provide a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb., and especially to provide an accurate and highly characteristic method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. and its application. The method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. provided by the present invention has high accuracy and strong characteristics, and is of great significance for the quality control of clinical Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. slices.

[0008] To achieve the purpose of this invention, the following technical solutions are adopted:

[0009] On the one hand, the present invention provides a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb., and the identification method includes the following steps:

[0010] (1) Mix the test sample with a solvent for reflux extraction, collect the filtrate, mix it with an organic solution and let it stand, then take the supernatant for desolvation, dissolve the residue with an organic solution and filter it to obtain the test solution A;

[0011] (2) Perform liquid chromatography detection on the test solution A obtained in step (1), compare the detection result with the characteristic chromatogram, and judge whether the test sample is Euryale ferox Salisb. or stir-fried Euryale ferox Salisb.

[0012] The above identification method can quickly and accurately identify the test sample by extracting the test sample, then performing liquid chromatography detection and comparing the results with the characteristic chromatogram. It has the characteristics of high accuracy and strong characteristics, and is of great significance for the quality control of clinical Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. slices; by using the method of desolvation and then redissolving with methanol solution, the peak shape of the peaks in the liquid chromatography detection result can be improved, and the response is better.

[0013] Preferably, before mixing the test sample with the solvent in step (1), the test sample is also pulverized and sieved.

[0014] Preferably, the sieve is a No. 3 sieve.

[0015] The above specific sieve can control the particle size of the test sample and improve the repeatability of the detection results.

[0016] Preferably, the solvent in step (1) includes water.

[0017] Preferably, the temperature of the reflux extraction in step (1) is 110 - 130 °C.

[0018] Preferably, the organic solution in step (1) includes methanol solution, and the volume fraction of the methanol solution is 50 - 100%.

[0019] Preferably, the flow rate of the mobile phase for the liquid chromatography detection in step (2) is 0.8 - 1.2 mL / min;

[0020] Preferably, the mobile phase for the liquid chromatography detection in step (2) comprises mobile phase A and mobile phase B. Mobile phase A is acetonitrile, and mobile phase B is a phosphoric acid solution with a volume fraction of 0.08 - 0.12%.

[0021] Among them, the temperature for reflux extraction can be 110°C, 112°C, 114°C, 116°C, 118°C, 120°C, 122°C, 124°C, 126°C, 128°C, 130°C, etc.; the volume fraction of the methanol solution can be 50%, 60%, 70%, 80%, 90%, 100%, etc.; 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 the phosphoric acid solution can be 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, etc. However, it is not limited to the values listed above, and other unlisted values within the above numerical ranges are equally applicable.

[0022] Preferably, the elution process for the liquid chromatography detection in step (2) is as follows: from 0 - 15 min, the volume fraction of mobile phase A changes uniformly from 0 to 2.8 - 3.2%, and the volume fraction of mobile phase B changes uniformly from 100% to 96.8 - 97.2%; from 15 - 20 min, the volume fraction of mobile phase A changes uniformly from 2.8 - 3.2% to 9.5 - 10.5%, and the volume fraction of mobile phase B changes uniformly from 96.8 - 97.2% to 89.5 - 90.5%; from 20 - 60 min, the volume fraction of mobile phase A changes uniformly from 9.5 - 10.5% to 17.5 - 18.5%, and the volume fraction of mobile phase B changes uniformly from 89.5 - 90.5% to 81.5 - 82.5%.

[0023] Among them, from 0 to 15 minutes, the volume fraction of mobile phase A can change to 2.8%, 2.9%, 3%, 3.1%, 3.2%, etc., and the volume fraction of mobile phase A can change to 96.8%, 96.9%, 97%, 97.1%, 97.2%, etc. From 15 to 20 minutes, the volume fraction of mobile phase A can change to 9.5%, 9.7%, 9.9%, 10.1%, 10.3%, 10.5%, etc., and the volume fraction of mobile phase B can change to 89.5%, 89.7%, 89.9%, 90.1%, 90.3%, 90.5%, etc. From 20 to 60 minutes, the volume fraction of mobile phase A can change to 17.5%, 17.7%, 17.9%, 18.1%, 18.3%, 18.5%, etc., and the volume fraction of mobile phase B can change to 81.5%, 81.7%, 81.9%, 82.1%, 82.3%, 82.5%, etc., but not limited to the values listed above. Other unlisted values within the above numerical range are equally applicable.

[0024] The above specific elution parameters can improve the resolution of the peaks in liquid chromatography and improve the detection accuracy.

[0025] Preferably, the chromatographic column for the liquid chromatography detection in step (2) is Shim-pack GIST C18-AQ.

[0026] Preferably, the characteristic chromatogram in step (2) is obtained by a method including the following steps:

[0027] (1’) Mix and reflux extract the semen euryales and stir-fried semen euryales with a solvent respectively, collect the filtrate, mix it with an organic solution and let it stand, then take the supernatant and remove the solvent, dissolve the residue with an organic solution and filter to obtain the test solution B;

[0028] (2’) Perform liquid chromatography detection on the test solution B obtained in step (1’) to obtain the detection results of semen euryales and stir-fried semen euryales;

[0029] (3’) Compare the detection results of semen euryales and stir-fried semen euryales obtained in step (2’), take the gallic acid peak as the reference peak (S peak) of the reference substance, and the gallic acid glucoside peak as the characteristic peak, and calculate the relative retention time or relative peak area of the characteristic peak.

[0030] The extraction methods of semen euryales and stir-fried semen euryales and the liquid chromatography detection parameters in the method for obtaining the above characteristic chromatogram are the same as those described above.

[0031] The characteristic chromatogram obtained by the above method can effectively show the distinguishing features of semen euryales and stir-fried semen euryales, and improve the accuracy and characteristic of the identification method of semen euryales and stir-fried semen euryales.

[0032] Preferably, the relative retention time in step (3’) is 1.15 - 1.27, such as 1.15, 1.16, 1.17, 1.18, 1.19, 1.20, 1.21, 1.22, 1.23, 1.24, 1.25, 1.26 or 1.27, etc., but not limited to the values listed above. Other unlisted values within the above numerical range are equally applicable.

[0033] Preferably, the comparison of the detection result with the characteristic chromatogram in step (2) is specifically as follows: Observe whether characteristic peaks appear in the detection result, calculate the relative retention time or relative peak area of the characteristic peaks, and then compare with the characteristic chromatogram to determine whether the sample to be tested is Euryale ferox Salisb. or stir-fried Euryale ferox Salisb.

[0034] On the other hand, the present invention also provides the application of the above-mentioned identification method of Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. in the identification of traditional Chinese medicine decoction pieces.

[0035] Compared with the prior art, the present invention has the following beneficial effects:

[0036] The present invention provides an identification method for Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. By extracting the sample to be tested, then detecting it by liquid chromatography and comparing the results with the characteristic chromatogram, it can quickly and accurately identify the sample to be tested, with high accuracy and strong characteristic. It has very important significance for the quality control of clinical Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. decoction pieces; using the method of desolvation and then redissolving with methanol solution can improve the peak shape of the peaks in the liquid chromatography detection result and have better response; by selecting a specific sieve, the particle size of the sample to be tested can be controlled, improving the repeatability of the detection result; by controlling specific elution parameters, the resolution of the peaks in the liquid chromatography can be improved, enhancing the detection accuracy; at the same time, the characteristic chromatogram obtained by a specific method can effectively show the distinguishing features between Euryale ferox Salisb. and stir-fried Euryale ferox Salisb., improving the accuracy and characteristic of the identification method for Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 is the detection result diagram in the preparation of the characteristic chromatogram;

[0038] Figure 2 is the detection result diagram of gallic acid sample in the preparation of the characteristic chromatogram;

[0039] Figure 3 is the detection result diagram of repeated injection of Euryale ferox Salisb. medicinal material;

[0040] Figure 4 is the detection result diagram of repeated injection of stir-fried Euryale ferox Salisb. medicinal material;

[0041] Figure 5 is the detection result diagram of Examples 1 - 3;

[0042] Figure 6 is the detection result diagram of Example 4;

[0043] Figure 7 It is the repeated detection result diagram of Example 1;

[0044] Figure 8 It is the detection result diagram of No. 4 sieve;

[0045] Figure 9 It is the detection result diagram without sieving. Detailed implementation manners

[0046] The technical solution of the present invention will be further described below through specific implementation manners. 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.

[0047] In the following examples, stir-fried Euryale ferox seeds are prepared by the following method:

[0048] Take the original Euryale ferox seeds medicinal materials, remove impurities, sieve through a 16-mesh sieve to remove drug residues, place the purified Euryale ferox seeds in a frying machine at a temperature of 260 °C and a rotation speed of 20 revolutions / min, and fry until slightly yellow to obtain the stir-fried Euryale ferox seeds.

[0049] Instruments used: Shimadzu high-performance liquid chromatography integrated instrument (LC-2030C 3D Plus); the chromatographic column is Shim-pack GIST C18-AQ (5 μm, 4.6×250 mm).

[0050] Preparation of characteristic chromatogram:

[0051] Take the Euryale ferox seeds medicinal materials and stir-fried Euryale ferox seeds medicinal materials, crush them respectively, sieve through a No. 3 sieve, take 10 g each, weigh accurately, place them in a stoppered conical flask, add 150 mL of pure water, reflux and extract for 1 hour with an electric heating mantle (set at 120 °C), filter while it is hot, collect the filtrate, add 100 mL of pure water to the medicinal residues and reflux and extract for 1 hour, filter, combine the two filtrates, and make up to 250 mL with water. After shaking well, measure 50 mL of the extract, add 50 mL of methanol, stir well, let stand for 10 minutes, transfer 50 mL of the supernatant, recover the solvent under reduced pressure at 60 °C, dissolve the residue in 10 mL of a 50% (v / v) methanol solution, filter through a 0.45 μm filter membrane, and take the filtrate to obtain the test solution. Perform liquid chromatography detection on the test solution, use acetonitrile as mobile phase A and a 0.1% (v / v) 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 270 nm, and the injection volume is 20 μL.

[0052]

[0053] The detection results are as Figure 1 shown. The main peaks are respectively denoted as 1-5. In addition, the gallic acid reference substance is detected according to the above method, and the results are asFigure 2 As shown. It can be found from the figure that the peak No. 1 is gallic acid, and it has a fast peak emergence and a large peak area. Therefore, it is selected as the S peak. In addition, the peak No. 5 only appears in the test results of stir-fried Euryale ferox seeds. Therefore, it is selected as the characteristic peak and its mass spectrometry characterization is carried out. The results are as follows:

[0054] Adduct ion [M-H]-, MS / MS data: 271.0438; 211.0254; 169.0126; 124.0154, the actual value of m / z is 331.065, which proves that it is galloyl glucoside. Then, the test results of repeated processing, injection and collection of different batches of Euryale ferox seed materials and stir-fried Euryale ferox seed materials are carried out (see Figures 3 - 4 ), and the relative retention time is calculated to be 1.15 - 1.27.

[0055] Example 1

[0056] This example provides a method for identifying Euryale ferox seeds and stir-fried Euryale ferox seeds. The specific steps are as follows:

[0057] Take the test sample, crush it, pass through a No. 3 sieve, take 10 g, weigh it accurately, place it in a stoppered conical flask, add 150 mL of pure water, reflux and extract for 1 hour with an electric heating mantle (set at 120 °C), filter while it is hot, collect the filtrate, add 100 mL of pure water to the residue and reflux and extract for 1 hour, filter, combine the two filtrates, and make up to 250 mL with water. After shaking well, measure 50 mL of the extract, add 50 mL of methanol, stir well, let it stand for 10 minutes, transfer 50 mL of the supernatant, recover the solvent under reduced pressure at 60 °C, dissolve the residue in 10 mL of a 50% (v / v) methanol solution, filter through a 0.45 μm filter membrane, and take the filtrate to obtain the test solution. Carry out liquid chromatography detection on the test solution, use acetonitrile as mobile phase A and a 0.1% (v / v) phosphoric acid solution as mobile phase B, and carry out 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 270 nm, and the injection volume is 20 μL.

[0058] Example 2

[0059] This example provides a method for identifying Euryale ferox seeds and stir-fried Euryale ferox seeds. The specific steps are as follows:

[0060] Take the sample to be tested, crush it, sieve it through a No. 3 sieve, take 10 g, weigh it accurately, place it in a stoppered conical flask, add 150 mL of pure water, reflux and extract for 1 hour with an electric heating mantle (set at 120 °C), filter while it is hot, collect the filtrate, add another 100 mL of pure water to the medicinal residue and reflux and extract for 1 hour, filter, combine the two filtrates, and make up to 250 mL with water. After shaking well, measure 50 mL of the extract, add 50 mL of methanol, stir well, let stand for 10 minutes, transfer 50 mL of the supernatant, filter through a 0.45 μm filter membrane, and take the filtrate 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 by volume 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 270 nm, and the injection volume is 20 μL.

[0061] Example 3

[0062] This example provides a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb., and the specific steps are as follows:

[0063] Take the sample to be tested, crush it, sieve it through a No. 3 sieve, take 10 g, weigh it accurately, place it in a stoppered conical flask, add 150 mL of pure water, reflux and extract for 1 hour with an electric heating mantle (set at 120 °C), filter while it is hot, collect the filtrate, add another 100 mL of pure water to the medicinal residue and reflux and extract for 1 hour, filter, combine the two filtrates, and make up to 250 mL with water. After shaking well, measure 50 mL of the extract, add 100 mL of methanol, stir well, let stand for 10 minutes, transfer 50 mL of the supernatant, filter through a 0.45 μm filter membrane, and take the filtrate 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 by volume 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 270 nm, and the injection volume is 20 μL.

[0064] Example 4

[0065] This example provides a method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. Except for the different samples to be tested, the rest are the same as in Example.

[0066] The test results of Examples 1 - 3 are as Figure 5 shown. It can be seen from the figure that the samples to be tested in Examples 1 - 3 do not contain characteristic peaks, so it is determined that they are all Euryale ferox Salisb.; at the same time, it can be seen from Figure 5 that by using the method of desolvation and then redissolving with methanol solution, compared with the method of only using alcohol precipitation, the peak shape and response of the chromatographic peaks are better in the present invention.

[0067] The test result of Example 4 is as Figure 6As shown. It can be seen from the figure that the characteristic peak of galloyl glucoside appears, and the relative retention time is calculated to be 1.2114. From this, it can be obtained that the sample to be tested in Example 4 is stir-fried Euryale ferox seeds.

[0068] In addition, the identification method of Example 1 was repeated five times, and five times were repeated respectively on the premise of changing the type of sieve (No. 4 sieve) and not sieving, with the rest unchanged. The results are as Figures 7 - 9 . It can be seen from the figure that by using the No. 3 sieve, the present invention significantly improves the repeatability of the detection results compared with other sieves.

[0069] The applicant declares that the present invention uses the above embodiments to illustrate the identification method and its application of Euryale ferox seeds and stir-fried Euryale ferox seeds of the present invention. However, 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 substitution 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.

[0070] 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 technical concept scope of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0071] In addition, it should be noted that in the various specific technical features described in the above specific embodiments, they can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb., characterized in that, the identification method comprises the following steps: (1) Crushing the sample to be tested, passing through a No. 3 sieve, mixing the sample to be tested with water and refluxing and extracting at 110-130 °C, collecting the filtrate, mixing it with an organic solution and standing, then taking the supernatant for desolvation, dissolving the residue with an organic solution, filtering to obtain the test solution A; (2) Performing liquid chromatography detection on the test solution A obtained in step (1), comparing the detection result with the characteristic chromatogram, and judging whether the sample to be tested is Euryale ferox Salisb. or stir-fried Euryale ferox Salisb.; The elution process of the liquid chromatography detection is as follows: from 0 to 15 min, the volume fraction of mobile phase A changes uniformly from 0 to 2.8-3.2%, and the volume fraction of mobile phase B changes uniformly from 100% to 96.8-97.2%; from 15 to 20 min, the volume fraction of mobile phase A changes uniformly from 2.8-3.2% to 9.5-10.5%, and the volume fraction of mobile phase B changes uniformly from 96.8-97.2% to 89.5-90.5%; from 20 to 60 min, the volume fraction of mobile phase A changes uniformly from 9.5-10.5% to 17.5-18.5%, and the volume fraction of mobile phase B changes uniformly from 89.5-90.5% to 81.5-82.5%; The flow rate of the mobile phase for the liquid chromatography detection is 0.8-1.2 mL / min; the mobile phase for the liquid chromatography detection includes mobile phase A and mobile phase B, mobile phase A is acetonitrile, mobile phase B is a phosphoric acid solution, and the volume fraction of the phosphoric acid solution is 0.08-0.12%; the chromatographic column for the liquid chromatography detection is Shim-pack GIST C18-AQ; The characteristic chromatogram is obtained by a method comprising the following steps: (1') Mixing Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. with a solvent respectively for refluxing and extracting, collecting the filtrate, mixing it with an organic solution and standing, then taking the supernatant for desolvation, dissolving the residue with an organic solution, filtering to obtain the test solution B; (2') Performing liquid chromatography detection on the test solution B obtained in step (1') to obtain the detection result of Euryale ferox Salisb. and the detection result of stir-fried Euryale ferox Salisb.; (3') Comparing the detection result of Euryale ferox Salisb. and the detection result of stir-fried Euryale ferox Salisb. obtained in step (2'), taking the gallic acid peak as the S peak and the galloyl glucoside peak as the characteristic peak of stir-fried Euryale ferox Salisb., and calculating the relative retention time or relative peak area of the characteristic peak.

2. The method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. according to claim 1, characterized in that, the organic solution in step (1) includes a methanol solution, and the volume fraction of the methanol solution is 50-100%.

3. The method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. according to claim 1, characterized in that, the relative retention time in step (3') is 1.15-1.

27.

4. The method for identifying Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. according to claim 1, characterized in that, the comparison of the detection result with the characteristic chromatogram in step (2) specifically is: observing whether a characteristic peak appears in the detection result, calculating the relative retention time or relative peak area of the characteristic peak, and then comparing it with the characteristic chromatogram to judge whether the sample to be tested is Euryale ferox Salisb. or stir-fried Euryale ferox Salisb.

5. Use of the identification method of Euryale ferox Salisb. and stir-fried Euryale ferox Salisb. according to any one of claims 1-4 in the identification of traditional Chinese medicine decoction pieces.

Images