Construction method and application of fingerprint spectrum of Chinese medicinal materials of Plantago asiatica, Plantago asiatica and mixed original Plantago asiatica

Abstract

This invention relates to the field of traditional Chinese medicine detection technology, and particularly to a method for constructing fingerprint spectra of Plantago asiatica, Plantago asiatica var. chinensis, and Plantago asiatica var. chinensis mixed origin, and their applications. The method for constructing the fingerprint spectra of Plantago asiatica var. chinensis includes the following steps: taking the whole herb or a predetermined part of Plantago asiatica var. chinensis, the predetermined part being the rhizome, leaf, or spike; performing ultra-high performance liquid chromatography (UHPLC) on the test sample solution; the chromatographic conditions include: mobile phase A comprising acetonitrile, and mobile phase B comprising an aqueous phosphoric acid solution, with gradient elution. The fingerprint spectra constructed by this invention can be used to solve the problem of distinguishing and identifying Plantago asiatica, Plantago asiatica var. chinensis, and commercially available mixed origin Plantago asiatica var. chinensis, as well as to analyze the quality of Plantago asiatica and clarify its origin.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, in particular to a method for constructing fingerprints of Plantago asiatica, Plantago asiatica and mixed-origin Plantago asiatica Chinese medicinal materials and applications thereof, specifically to a method for constructing fingerprints of Plantago asiatica Chinese medicinal materials, a method for constructing fingerprints of Plantago asiatica Chinese medicinal materials, a method for constructing fingerprints of mixed-origin Plantago asiatica Chinese medicinal materials, a method for identifying different parts of Plantago asiatica Chinese medicinal materials, a method for identifying different parts of Plantago asiatica Chinese medicinal materials, a method for identifying Plantago asiatica Chinese medicinal materials, Plantago asiatica Chinese medicinal materials and mixed-origin Plantago asiatica Chinese medicinal materials, and a method for evaluating the quality of Plantago asiatica Chinese medicinal materials, Plantago asiatica Chinese medicinal materials and mixed-origin Plantago asiatica Chinese medicinal materials. Background Art

[0002] Plantain is the dried whole herb of Plantago asiatica L. or Plantago depressa Willd. of the Plantaginaceae family. It is cold in nature and sweet in taste. It has the effects of clearing away heat, promoting diuresis, relieving stranguria, removing phlegm, cooling blood, and detoxifying. Plantain mainly contains phenylethanol glycosides such as plantainoside, verbascoside, isoverbascoside, and plantagoside D, flavonoids such as luteolin, luteolin, and apigenin and their glycosides, as well as iridoids such as geniposide and aucubin and their glycosides. These chemical components have pharmacological effects such as antiviral, tumor, and immunity enhancement, anti-oxidation, and neuroprotection. However, the 2020 edition of the "Chinese Pharmacopoeia" only stipulates the limit of plantainoside under the item of plantain content determination. This single indicator component is obviously difficult to fully reflect the overall quality of plantain materials.

[0003] In addition, the herbal materials of plantain circulating on the market often have the situation of mixing the source of plantain and plantain, which is difficult to distinguish simply from the properties. The characteristic spectrum / fingerprint spectrum of traditional Chinese medicine is a chemical representation of the overall nature of traditional Chinese medicine, and is widely used in the quality evaluation of traditional Chinese medicine. Therefore, by constructing the common peak patterns of the fingerprints of plantain, plantain and commercially mixed original herbal materials of plantain, the situation of mixing the source of plantain and plantain on the market can be solved, and the problem of their uneven quality can be solved. However, the common peaks of the characteristic spectrum / fingerprint spectrum of the plantago medicinal materials currently constructed are less calibrated and identified, making it difficult to distinguish between plantain, plantain and commercially mixed original herbal materials of plantain. Summary of the invention

[0004] Based on this, the first aspect of the present invention provides a method for constructing a fingerprint of a Chinese medicinal material Plantago asiatica. The technical solution is as follows:

[0005] A method for constructing a fingerprint spectrum of a Chinese medicinal material Plantago asiatica comprises the following steps:

[0006] The whole plant or a predetermined part of the Chinese medicinal material Plantago asiatica is taken, crushed, and then an extraction solvent is added for extraction treatment, and the extract is collected to prepare a test solution, wherein the predetermined part is a rhizome, a leaf or an ear;

[0007] Taking the test solution for ultra-high performance liquid chromatography determination;

[0008] The chromatographic conditions of the ultra-high performance liquid chromatography include:

[0009] The mobile phase includes mobile phase A and mobile phase B, wherein the mobile phase A includes acetonitrile, and the mobile phase B includes a phosphoric acid aqueous solution, and the elution is performed by gradient elution;

[0010] The gradient elution includes the following procedures:

[0011] 0-2min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%;

[0012] 2min to 4min, the volume fraction of the mobile phase A increases from 10% to 13%, and the volume fraction of the mobile phase B decreases from 90% to 87%;

[0013] 4min to 14min, the volume fraction of the mobile phase A is maintained at 13%, and the volume fraction of the mobile phase B is maintained at 87%;

[0014] From 14min to 24min, the volume fraction of the mobile phase A increased from 13% to 15%, and the volume fraction of the mobile phase B decreased from 87% to 85%;

[0015] From 24 to 25 minutes, the volume fraction of the mobile phase A increases from 15% to 17%, and the volume fraction of the mobile phase B decreases from 85% to 83%;

[0016] From 25min to 31min, the volume fraction of the mobile phase A increased from 17% to 22%, and the volume fraction of the mobile phase B decreased from 83% to 78%;

[0017] From 31min to 41min, the volume fraction of the mobile phase A increases from 22% to 40%, and the volume fraction of the mobile phase B decreases from 78% to 60%;

[0018] From 41min to 50min, the volume fraction of the mobile phase A increases from 40% to 68%, and the volume fraction of the mobile phase B decreases from 60% to 32%;

[0019] From 50min to 53min, the volume fraction of the mobile phase A increases from 68% to 88%, and the volume fraction of the mobile phase B decreases from 32% to 12%;

[0020] From 53min to 54min, the volume fraction of the mobile phase A is reduced from 88% to 10%, and the volume fraction of the mobile phase B is increased from 12% to 90%;

[0021] From 54 min to 60 min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%.

[0022] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography further include:

[0023] 0-3min, the detection wavelength is 240nm-250nm; 3min-48min, the detection wavelength is 320nm-340nm, 48min-60min, the detection wavelength is 240nm-250nm.

[0024] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a column temperature of 28°C to 32°C.

[0025] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a flow rate of 0.25 mL / min to 0.35 mL / min.

[0026] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: an injection volume of 1 μL to 2 μL.

[0027] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: using octadecylsilane bonded silica gel as a filler.

[0028] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: the volume fraction of phosphoric acid in the phosphoric acid aqueous solution is 0.05% to 0.2%.

[0029] In one embodiment, the extraction solvent is methanol-water solution.

[0030] In one embodiment, 40 mL to 60 mL of the extraction solvent is added for every 1 gram of the whole plant or predetermined part of the Chinese medicinal material Plantago asiatica.

[0031] In one embodiment, the extraction process is ultrasonic treatment.

[0032] In one of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0033] In one embodiment, the ultrasonic treatment time is 20 min to 40 min.

[0034] The second aspect of the present invention provides a method for constructing a fingerprint spectrum of a Chinese medicinal material Plantago asiatica. The technical scheme is as follows:

[0035] A method for constructing a fingerprint spectrum of a Chinese medicinal material of Plantago asiatica comprises the following steps:

[0036] The whole plant or a predetermined part of the Chinese medicinal material Plantago asiatica is taken, crushed, and then an extraction solvent is added for extraction treatment, and the extract is collected to prepare a test solution, wherein the predetermined part is a rhizome, a leaf or an ear;

[0037] Taking the test solution for ultra-high performance liquid chromatography determination;

[0038] The chromatographic conditions of the ultra-high performance liquid chromatography include:

[0039] The mobile phase includes mobile phase A and mobile phase B, wherein the mobile phase A includes acetonitrile, and the mobile phase B includes a phosphoric acid aqueous solution, and the elution is performed by gradient elution;

[0040] The gradient elution includes the following procedures:

[0041] 0-2min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%;

[0042] 2min to 4min, the volume fraction of the mobile phase A increases from 10% to 13%, and the volume fraction of the mobile phase B decreases from 90% to 87%;

[0043] 4min to 14min, the volume fraction of the mobile phase A is maintained at 13%, and the volume fraction of the mobile phase B is maintained at 87%;

[0044] From 14min to 24min, the volume fraction of the mobile phase A increased from 13% to 15%, and the volume fraction of the mobile phase B decreased from 87% to 85%;

[0045] From 24 to 25 minutes, the volume fraction of the mobile phase A increases from 15% to 17%, and the volume fraction of the mobile phase B decreases from 85% to 83%;

[0046] From 25min to 31min, the volume fraction of the mobile phase A increased from 17% to 22%, and the volume fraction of the mobile phase B decreased from 83% to 78%;

[0047] From 31min to 41min, the volume fraction of the mobile phase A increases from 22% to 40%, and the volume fraction of the mobile phase B decreases from 78% to 60%;

[0048] From 41min to 50min, the volume fraction of the mobile phase A increases from 40% to 68%, and the volume fraction of the mobile phase B decreases from 60% to 32%;

[0049] From 50min to 53min, the volume fraction of the mobile phase A increases from 68% to 88%, and the volume fraction of the mobile phase B decreases from 32% to 12%;

[0050] From 53min to 54min, the volume fraction of the mobile phase A is reduced from 88% to 10%, and the volume fraction of the mobile phase B is increased from 12% to 90%;

[0051] From 54 min to 60 min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%.

[0052] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography further include:

[0053] 0-3min, the detection wavelength is 240nm-250nm; 3min-48min, the detection wavelength is 320nm-340nm, 48min-60min, the detection wavelength is 240nm-250nm.

[0054] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a column temperature of 28°C to 32°C.

[0055] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a flow rate of 0.25 mL / min to 0.35 mL / min.

[0056] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: an injection volume of 1 μL to 2 μL.

[0057] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: using octadecylsilane bonded silica gel as a filler.

[0058] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: the volume fraction of phosphoric acid in the phosphoric acid aqueous solution is 0.05% to 0.2%.

[0059] In one embodiment, the extraction solvent is methanol-water solution.

[0060] In one embodiment, 40 mL to 60 mL of the extraction solvent is added to every 1 g of the whole plant or predetermined part of the Plantago Chinese medicinal material.

[0061] In one embodiment, the extraction process is ultrasonic treatment.

[0062] In one of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0063] In one embodiment, the ultrasonic treatment time is 20 min to 40 min.

[0064] The third aspect of the present invention provides a method for constructing a fingerprint of a mixed-origin Chinese medicinal material of Plantago asiatica, and the technical scheme thereof is as follows:

[0065] A method for constructing a fingerprint spectrum of a mixed-origin plantain Chinese medicinal material comprises the following steps:

[0066] Take the mixed original plantain Chinese medicinal material, crush it, add the extraction solvent to perform extraction treatment, collect the extract, and prepare the test solution;

[0067] Taking the test solution for ultra-high performance liquid chromatography determination;

[0068] The chromatographic conditions of the ultra-high performance liquid chromatography include:

[0069] The mobile phase includes mobile phase A and mobile phase B, wherein the mobile phase A includes acetonitrile, and the mobile phase B includes a phosphoric acid aqueous solution, and the elution is performed by gradient elution;

[0070] The gradient elution includes the following procedures:

[0071] 0-2min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%;

[0072] 2min to 4min, the volume fraction of the mobile phase A increases from 10% to 13%, and the volume fraction of the mobile phase B decreases from 90% to 87%;

[0073] 4min to 14min, the volume fraction of the mobile phase A is maintained at 13%, and the volume fraction of the mobile phase B is maintained at 87%;

[0074] From 14min to 24min, the volume fraction of the mobile phase A increased from 13% to 15%, and the volume fraction of the mobile phase B decreased from 87% to 85%;

[0075] From 24 to 25 minutes, the volume fraction of the mobile phase A increases from 15% to 17%, and the volume fraction of the mobile phase B decreases from 85% to 83%;

[0076] From 25min to 31min, the volume fraction of the mobile phase A increased from 17% to 22%, and the volume fraction of the mobile phase B decreased from 83% to 78%;

[0077] From 31min to 41min, the volume fraction of the mobile phase A increases from 22% to 40%, and the volume fraction of the mobile phase B decreases from 78% to 60%;

[0078] From 41min to 50min, the volume fraction of the mobile phase A increases from 40% to 68%, and the volume fraction of the mobile phase B decreases from 60% to 32%;

[0079] From 50min to 53min, the volume fraction of the mobile phase A increases from 68% to 88%, and the volume fraction of the mobile phase B decreases from 32% to 12%;

[0080] From 53min to 54min, the volume fraction of the mobile phase A is reduced from 88% to 10%, and the volume fraction of the mobile phase B is increased from 12% to 90%;

[0081] From 54 min to 60 min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%.

[0082] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography further include:

[0083] 0-3min, the detection wavelength is 240nm-250nm; 3min-48min, the detection wavelength is 320nm-340nm, 48min-60min, the detection wavelength is 240nm-250nm.

[0084] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a column temperature of 28°C to 32°C.

[0085] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: a flow rate of 0.25 mL / min to 0.35 mL / min.

[0086] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: an injection volume of 1 μL to 2 μL.

[0087] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: using octadecylsilane bonded silica gel as a filler.

[0088] In one embodiment, the chromatographic conditions of the ultra-high performance liquid chromatography also include: the volume fraction of phosphoric acid in the phosphoric acid aqueous solution is 0.05% to 0.2%.

[0089] In one embodiment, the extraction solvent is methanol-water solution.

[0090] In one embodiment, 40 mL to 60 mL of the extraction solvent is added for every 1 g of the mixed-base original plantain Chinese medicinal material.

[0091] In one embodiment, the extraction process is ultrasonic treatment.

[0092] In one of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0093] In one embodiment, the ultrasonic treatment time is 20 min to 40 min.

[0094] The fourth aspect of the present invention provides a method for identifying different parts of a Chinese medicinal material, Plantago asiatica, and the technical solution thereof is as follows:

[0095] A method for identifying different parts of a Chinese medicinal material, Plantago asiatica, comprises the following steps:

[0096] Taking the part of the Chinese medicinal material Plantago asiatica to be tested, crushing it, adding an extraction solvent to perform extraction treatment, collecting the extract, and preparing a sample solution to be tested;

[0097] According to the above-mentioned chromatographic conditions, the sample solution to be tested is subjected to ultra-high performance liquid chromatography, the obtained chromatogram is matched with the fingerprint spectrum of the above-mentioned preset part, the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded, and the attribution of the part to be tested is determined.

[0098] In one embodiment, the identification criteria include:

[0099] The characteristic peak of verbascoside was recorded as peak 17, which was used as a reference peak. The characteristic peak with a retention time ratio of 0.943 to 0.950 to peak 17 was recorded as peak 16, and the characteristic peak with a retention time ratio of 1.350 to 1.360 to peak 17 was recorded as peak 21.

[0100] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica shows a characteristic peak with a retention time consistent with the fingerprint of the rhizome, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.006-0.029; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0-0.009, then the part to be tested is determined to be the rhizome;

[0101] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.001 to 0.004; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.002 to 0.011, then the part to be tested is determined to be the ear;

[0102] If the chromatogram of the tested part of the Chinese medicinal material Plantago asiatica shows a characteristic peak with a retention time consistent with the above-mentioned fingerprint spectrum of the leaves, and meets the following conditions at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.002-0.005; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.012-0.024, then the tested part is determined to be the leaf.

[0103] In one embodiment, the extraction solvent is methanol-water solution.

[0104] In one embodiment, 40 mL to 60 mL of the extraction solvent is added to each 1 g of the tested part of the Chinese medicinal material Plantago asiatica.

[0105] In one embodiment, the extraction process is ultrasonic treatment.

[0106] In one of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0107] In one embodiment, the ultrasonic treatment time is 20 min to 40 min.

[0108] The fifth aspect of the present invention provides a method for identifying different parts of a Chinese medicinal material of Plantago, and the technical scheme thereof is as follows:

[0109] A method for identifying different parts of a Chinese medicinal material of Plantago asiatica comprises the following steps:

[0110] Taking the part of the Chinese medicinal material of Plantago asiatica to be tested, crushing it, adding an extraction solvent to perform extraction treatment, collecting the extract, and preparing a sample solution to be tested;

[0111] According to the above-mentioned chromatographic conditions, the sample solution to be tested is subjected to ultra-high performance liquid chromatography, the obtained chromatogram is matched with the fingerprint spectrum of the above-mentioned preset part, the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded, and the part to be tested is determined.

[0112] In one embodiment, the identification criteria include:

[0113] The characteristic peak of plantagoside D is recorded as peak 18, and with it as the reference peak, the characteristic peak with a retention time ratio of 0.944 to 0.966 to peak 18 is recorded as peak 17 (verbascoside), and the characteristic peak with a retention time ratio of 1.272 to 1.286 to peak 18 is recorded as peak 21;

[0114] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the rhizome, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0-0.392; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.226-0.655, then the part to be tested is determined to be the rhizome;

[0115] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 1.119-3.312; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.228-1.076, then the part to be tested is determined to be the ear;

[0116] If the chromatogram of the tested part of the Plantago Chinese medicinal material shows a characteristic peak with a retention time consistent with the above-mentioned fingerprint spectrum of the leaves, and at the same time meets the following conditions: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0.091~0.671; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.676~5.169, then the tested part is determined to be the leaf.

[0117] In some embodiments, the extraction solvent is methanol-water solution.

[0118] In some embodiments, 40 mL to 60 mL of the extraction solvent is added for every 1 g of the tested part of the Plantago Chinese medicinal material.

[0119] In some embodiments, the extraction process is ultrasonic treatment.

[0120] In some of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0121] In some embodiments, the ultrasonic treatment time is 20 min to 40 min.

[0122] The sixth aspect of the present invention provides a method for identifying Chinese medicinal materials including Plantago asiatica, Plantago asiatica and mixed-origin Plantago asiatica, and the technical scheme thereof is as follows:

[0123] A method for identifying Chinese medicinal materials of Plantago asiatica, Chinese medicinal materials of Plantago asiatica and Chinese medicinal materials of mixed origin of Plantago asiatica comprises the following steps:

[0124] Take the Chinese medicinal materials to be tested, crush them, add extraction solvent to perform extraction treatment, collect the extract, and prepare the sample solution to be tested;

[0125] According to the above-mentioned chromatographic conditions, or the above-mentioned chromatographic conditions, or the above-mentioned chromatographic conditions, the sample solution to be tested is subjected to ultra-high performance liquid chromatography determination, the obtained chromatogram is matched with the fingerprint spectrum of the whole plant of the above-mentioned Plantago asiatica Chinese medicinal material, the fingerprint spectrum of the whole plant of the above-mentioned Plantago asiatica Chinese medicinal material and the fingerprint spectrum of the above-mentioned mixed-base Plantago asiatica Chinese medicinal material, the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded, and the type of the Chinese medicinal material to be tested is determined.

[0126] In some embodiments, the identification criteria include:

[0127] The characteristic peak of plantagoside is recorded as peak 11, and the characteristic peak with a retention time ratio of 1.346 to 1.372 with respect to peak 11 is recorded as peak 17 (verbascoside);

[0128] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the whole herb of the Chinese medicinal material Plantago asiatica, and the ratio of the peak area of ​​peak 17 to peak 11 is 48.799-152.931, it is determined that the Chinese medicinal material to be tested is the whole herb of the Chinese medicinal material Plantago asiatica;

[0129] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the whole herb of the Chinese medicinal material Plantago, and the ratio of the peak area of ​​peak 17 to peak 11 is 0.013-0.055, then it is determined that the Chinese medicinal material to be tested is the whole herb of the Chinese medicinal material Plantago;

[0130] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the above-mentioned mixed-base original plantain Chinese medicinal material, and the ratio of the peak area of ​​peak 17 to peak 11 is 0.148-2.326, then it is determined that the Chinese medicinal material to be tested is a mixed-base original plantain Chinese medicinal material.

[0131] In some embodiments, the extraction solvent is methanol-water solution.

[0132] In some embodiments, 40 mL to 60 mL of the extraction solvent is added for every 1 g of the Chinese medicinal material to be tested.

[0133] In some embodiments, the extraction process is ultrasonic treatment.

[0134] In some of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0135] In some embodiments, the ultrasonic treatment time is 20 min to 40 min.

[0136] The seventh aspect of the present invention provides a method for evaluating the quality of Chinese medicinal materials including Plantago asiatica, Plantago asiatica and mixed-origin Plantago asiatica. The technical scheme is as follows:

[0137] A method for evaluating the quality of Chinese medicinal materials including plantain, plantain and mixed plantago asiatica, comprising the following steps:

[0138] A group of Chinese medicinal materials to be tested are taken, crushed respectively, and then an extraction solvent is added for extraction treatment, and the extract is collected to prepare a group of sample solutions to be tested, wherein the group of Chinese medicinal materials to be tested includes one or more of the Chinese medicinal materials of Plantago asiatica to be tested, the Chinese medicinal materials of Plantago asiatica to be tested, and the Chinese medicinal materials of mixed-base Plantago asiatica to be tested;

[0139] According to the above chromatographic conditions, or the above chromatographic conditions, or the above chromatographic conditions, respectively, a group of the sample solutions to be tested are subjected to ultra-high performance liquid chromatography determination, and the obtained chromatograms are matched with the fingerprint spectrum of the whole plant of the above-mentioned Chinese medicinal material Plantago asiatica, the fingerprint spectrum of the whole plant of the above-mentioned Chinese medicinal material Plantago asiatica, and the fingerprint spectrum of the above-mentioned mixed-base original Chinese medicinal material Plantago asiatica, and the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded;

[0140] According to formula (1), the peak area value x of the characteristic peak ij Processing is performed to obtain the numerical matrix R ij , the logarithmic matrix R ij Calculate the standard deviation and get the volatility value S j ; The peak area value x of the characteristic peak ij Imported into SPSS25.0, the correlation matrix was obtained, and according to formula (3), the conflict value A was obtained. j ; According to formula (4), calculate the information value C j ; According to formula (5), calculate the weight coefficient W j According to formula (6), the peak area value x of the characteristic peak i (j) Perform averaging to obtain the dimensionless unit sequence x' i’ (j), taking the maximum value unit sequence as the optimal reference sequence x0(j), the correlation coefficient ξ is calculated according to formula (7) i (j); Calculate the comprehensive evaluation result r according to formula (8) i ;

[0141]

[0142]

[0143] C j =S j ×A j (4)

[0144]

[0145]

[0146]

[0147]

[0148] Among them, i represents the batch of plantain, j represents the chromatographic peak, r represents the Pearson correlation coefficient, and ρ is 0.5;

[0149] r i The larger the value, the better the quality of the Chinese medicinal materials to be tested.

[0150] In some embodiments, the extraction solvent is methanol-water solution.

[0151] In some embodiments, 40 mL to 60 mL of the extraction solvent is added for every 1 g of the Chinese medicinal material to be tested.

[0152] In some embodiments, the extraction process is ultrasonic treatment.

[0153] In some of the embodiments, the volume fraction of methanol in the methanol aqueous solution is 40% to 80%.

[0154] In some embodiments, the ultrasonic treatment time is 20 min to 40 min.

[0155] Compared with the traditional solution, the present invention has the following beneficial effects:

[0156] According to the characteristics of the differences in components contained in different sources of plantain, the present invention reasonably controls the chromatographic conditions of ultra-high performance liquid chromatography, and respectively constructs fingerprints of different parts of plantain, different parts of plantain and mixed original plantain medicinal materials. The constructed fingerprint can be used to solve the problem of distinguishing and identifying plantain, plantain and commercially available mixed original plantain medicinal materials, and can also be used to analyze the quality of plantain and clarify the source of plantain. It is more targeted, the analysis method is more objective and comprehensive, and has guiding value for comprehensively reflecting the overall quality of plantain medicinal materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0157] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0158] Figure 1 This is the chromatogram overlay of 6 batches of Plantago asiatica (whole herb);

[0159] Figure 2 This is the chromatogram overlay of 6 batches of Plantago asiatica (rhizome);

[0160] Figure 3 This is the chromatogram overlay of 6 batches of Plantago asiatica (leaves);

[0161] Figure 4 This is the chromatogram overlay of 6 batches of Plantago asiatica (ear);

[0162] Figure 5 It is the comparative fingerprint of the whole herb, rhizome, leaf and spike of Plantago asiatica;

[0163] Figure 6 This is the chromatogram overlay of 7 batches of Plantago asiatica (whole herb);

[0164] Figure 7 This is the chromatogram overlay of 7 batches of Plantago asiatica (rhizome);

[0165] Figure 8 This is the chromatogram overlay of 7 batches of Plantago asiatica (leaves);

[0166] Fig. 9 This is the chromatogram overlay of 7 batches of Plantago asiatica (ear);

[0167] Fig.10 It is the comparative fingerprint of the whole herb, rhizome, leaf and spike of Plantago asiatica;

[0168] Fig.11 The chromatograms of 8 batches of mixed-origin plantain are overlaid;

[0169] Fig.12 The fingerprints of the plantago asiatica (whole herb), the plantago asiatica (whole herb) and the mixed-origin plantago asiatica are compared.

[0170] Fig.13 It is the chromatogram of the mixed reference substance (peak 1: geniposide, peak 2: 1-caffeoylquinic acid, peak 11: plantagoside, peak 13: luteolin, peak 17: verbascoside; peak 18: plantagoside D; peak 19: isoverbascoside; peak 20: apigenin-7-O-glucuronide; peak 24: luteolin; peak 25: apigenin);

[0171] Fig.14 The PCA score graph of Plantago asiatica (whole herb), Plantago asiatica (whole herb) and mixed origin;

[0172] Fig.15 The HCA graphs of Plantago asiatica (whole herb), Plantago asiatica (whole herb) and mixed-origin Plantago asiatica;

[0173] Fig.16 The PLS-DA graphs of Plantago asiatica (whole herb), Plantago asiatica (whole herb) and mixed-origin Plantago asiatica;

[0174] Fig.17 This is the VIP value chart of Plantago asiatica (whole herb), Plantago asiatica (whole herb) and mixed origin. DETAILED DESCRIPTION

[0175] The present invention will be further described in detail below in conjunction with specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thoroughly understood.

[0176] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0177] 1. Instruments and Materials

[0178] 1.1 Instrument

[0179] Thermo Fisher ultra-high performance liquid chromatograph (Waters, USA); KQ-700DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); Milli-Q Direct ultrapure water system (Merck Co., Ltd.); ME204E 1 / 10,000 balance (METTLERTOLEDO); DHG-9147A electric constant temperature drying oven (Shanghai Jinghong Experimental Equipment Co., Ltd.).

[0180] 1.2 Materials

[0181] Reference substances: geniposide (batch number 111828-201805, mass fraction, 98.1%); plantain glycoside (batch number 111914-202105, mass fraction ≥ 96.0%); luteolin (batch number 111720-202111, mass fraction ≥ 96.6%); verbascoside (batch number 11530-201914, mass fraction ≥ 95.2%); luteolin (batch number 111520-202006, mass fraction ≥ 94.4%); apigenin (batch number 111901-202004, mass fraction ≥ 99. 4%) were purchased from China Food and Drug Inspection Institute; 1-caffeoylquinic acid (batch number DST202101, mass score ≥98%) was purchased from Lotte Beauty Pharmaceutical Co., Ltd.; plantagoside D (batch number CFS202101, mass score ≥98.0%) was purchased from Wuhan Tianzhi Biotechnology Co., Ltd.; isoverbascoside (batch number 19092702, mass score ≥92.6%) was purchased from Chengdu Gelip Biotechnology Co., Ltd.; apigenin-7-O-glucuronide (batch number 21062901, mass score ≥98.3%) was purchased from Chengdu Pufeide Biotechnology Co., Ltd.

[0182] Methanol (analytical grade, Tianjin Fuyu Fine Chemical Co., Ltd.), acetonitrile (chromatographic grade, Merck Co., Ltd.), phosphoric acid (chromatographic grade, Foshan Xilong Chemical Co., Ltd.), and water were ultrapure water (obtained from the laboratory Milli-Q ultrapure water system).

[0183] There were 21 batches of plantain herbs, and the plantain herbs were identified by Wei Mei, chief pharmacist of Guangdong Yifang Pharmaceutical Co., Ltd. as the dried whole herbs of Plantago asiatica L. or Plantago depressa Willd. of the Plantaginaceae family. The collected herbs were 6 batches of plantain, 7 batches of plantain, and 8 batches of mixed original plantain. Among them, the plantain and plantain herbs were further separated according to the three parts of rhizome, ear and leaf, and samples of four parts including whole herb, rhizome, ear and leaf were obtained. The specific medicinal material information table is shown in Table 1.

[0184] Table 1

[0185]

[0186]

[0187] 2 Methods and Results

[0188] 2.1 Construction method of fingerprint spectrum of different parts of Chinese medicinal materials of Plantago asiatica, construction method of fingerprint spectrum of different parts of Chinese medicinal materials of Plantago asiatica, construction method of fingerprint spectrum of mixed original Chinese medicinal materials of Plantago asiatica

[0189] 2.1.1 Chromatographic conditions

[0190] Chromatographic column: Aglient SB C18 chromatographic column (2.1×100 mm, 1.6 μm); mobile phase: acetonitrile (mobile phase A)-0.1% phosphoric acid aqueous solution (mobile phase B), gradient elution: 0-2 min, 10% (A); 2-4 min, 10%-13% (A); 4-14 min, 13% (A); 14-24 min, 13%-15% (A); 24-25 min, 15%-17% (A); 25-31 min, 17%-22% (A ); 31-41min, 22%-40% (A); 41-50min, 40%-68% (A); 50-53min, 68%-88% (A); 53-54min, 88%-10% (A); 54-60min, 10% (A); detection wavelength: 0-3min, 245nm; 3-48min, 330nm; 48-60min, 245nm; column temperature 30℃; flow rate 0.3mL / min; injection volume 1μL.

[0191] 2.1.2 Preparation of reference solution

[0192] Take appropriate amount of geniposide, 1-caffeoylquinic acid, plantagoside, luteolin, verbascoside, plantagoside D, isoverbosaside, apigenin-7-O-glucuronide, luteolin, and apigenin reference substance, accurately weigh, and add 60% methanol aqueous solution to make a solution containing 24.66 μg geniposide, 17.93 μg 1-caffeoylquinic acid, 48. A mixed reference solution of 91 μg, 30.64 μg of luteolin, 62.60 μg of verbascoside, 64.97 μg of plantagoside D, 41.01 μg of isovorascoside, 43.06 μg of apigenin-7-O-glucuronide, 18.52 μg of luteolin and 14.23 μg of apigenin was filtered through a 0.22 μm microporous filter membrane, and the filtrate was taken to obtain the reference solution.

[0193] 2.1.3 Preparation of test solution

[0194] Take this product and grind it into fine powder (pass through No. 2 sieve), take 1.0g, weigh it accurately, add 50mL of 60% methanol aqueous solution by volume, weigh the mass, ultrasonicate for 30 minutes, take it out, put it to room temperature, make up the lost mass with 60% methanol aqueous solution by volume, shake well, filter, and take the filtrate to obtain the test solution.

[0195] 2.1.4 Methodological investigation

[0196] Prepare the test solution according to the method under "2.1.3", and perform the injection and determination according to the chromatographic conditions under "2.1.1", and examine the precision, repeatability and stability respectively.

[0197] The results showed that under various tests, the RSD value of the relative retention time of each characteristic peak was less than 1%, and the RSD value of the relative peak area of ​​each characteristic peak was less than 5%, indicating that the method is objective, accurate and reliable.

[0198] 2.1.5 Fingerprint calibration

[0199] Take the whole herb, leaves, rhizomes and spikes of Plantago asiatica from batches P1 to P6, prepare the test solution according to the method in "2.1.3", and inject and measure according to the chromatographic conditions in "2.1.1" to obtain chromatograms of 24 samples. Among them, the chromatograms of 6 batches of Plantago asiatica (whole herb) are as follows: Figure 1 As shown in Figure 2, there are 24 common peaks in total. The chromatogram overlay of 6 batches of Plantago asiatica (rhizome) is shown in Figure 2. Figure 2 As shown in FIG. 1 , there are 16 common peaks in total; the chromatogram overlay of 6 batches of Plantago asiatica (leaves) is shown in FIG. Figure 3 As shown in FIG. 1 , there are 23 common peaks in total. The chromatogram overlay of 6 batches of Plantago asiatica (ears) is shown in FIG. Figure 4 As shown, a total of 22 common peaks were marked.

[0200] The comparative fingerprints of the whole herb, rhizome, leaf and spike of Plantago asiatica are shown in Figure 5 .

[0201] Take the whole herb, leaves, rhizomes and spikes of Plantago from batches C1 to C7, prepare the test solution according to the method in "2.1.3", and inject and measure according to the chromatographic conditions in "2.1.1" to obtain chromatograms of 28 samples. Among them, the chromatograms of 7 batches of Plantago (whole herb) are as follows: Figure 6 As shown in Figure 2, there are 22 common peaks in total. The chromatogram overlay of 7 batches of Plantago asiatica (rhizome) is shown in Figure 2. Figure 7 As shown in FIG. 1 , there are 10 common peaks in total; the chromatogram overlay of 7 batches of Plantago asiatica (leaves) is shown in FIG. Figure 8 As shown in FIG. 1 , there are 22 common peaks in total. The chromatogram overlay of 7 batches of Plantago asiatica (ears) is shown in FIG. Fig. 9 As shown, a total of 16 common peaks were marked.

[0202] The comparative fingerprints of the whole herb, rhizome, leaf and spike of Plantago asiatica are shown in Fig.10 .

[0203] Take the whole plant of mixed origin plantain from batches H1 to H8, prepare the test solution according to the method under "2.1.3", inject and measure according to the chromatographic conditions under "2.1.1", and obtain chromatograms of 8 samples. Among them, the chromatograms of the 8 batches of mixed origin plantain are as follows: Fig.11 As shown, a total of 23 common peaks were marked.

[0204] The comparative fingerprints of Plantago asiatica (whole herb), Plantago asiatica (whole herb) and mixed-origin Plantago asiatica are shown in Fig.12 .

[0205] See the chromatogram of the mixed reference substance for Fig.13 A total of 10 chromatographic peaks were identified, namely peak 1: geniposide, peak 2: 1-caffeoylquinic acid, peak 11: plantagoside, peak 13: luteolin, peak 17: verbascoside; peak 18: plantagoside D; peak 19: isoverbascoside; peak 20: apigenin-7-O-glucuronide; peak 24: luteolin; peak 25: apigenin.

[0206] The similarities of the six batches of Plantago (whole herb) were 1.000, 0.995, 0.997, 0.989, 0.992, and 0.998, respectively; the similarities of the six batches of Plantago (rhizomes) were 1.000, 0.990, 0.990, 0.985, 0.993, and 0.991, respectively; the similarities of the six batches of Plantago (leaves) were 1.000, 0.993, 0.996, 0.995, 0.994, and 0.995, respectively; the similarities of the six batches of Plantago (ears) were 1.000, 0.985, 0.997, 0.996, 0.996, and 0.998, respectively. The similarities of the seven batches of Plantago (whole herb) were 1.000, 0.993, 0.997, 0.991, 0.972, The similarities of the seven batches of Plantain (rhizomes) were 1.000, 0.982, 0.998, 0.995, 0.977, 0.986 and 0.978 respectively; the similarities of the seven batches of Plantain (leaves) were 1.000, 0.998, 0.994, 0.993, 0.918, 0.913 and 0.988 respectively; the similarities of the seven batches of Plantain (ears) were 1.000, 0.906, 0.980, 0.963, 0.922, 0.998 and 0.915 respectively. The similarities of the eight mixed batches of Plantain (whole herb) were 1.000, 0.867, 0.941, 0.961, 0.910, 0.859, 0.926 and 0.930 respectively. Among them, the similarity results between batches of different medicinal parts with different origins were all >0.91, indicating that the chemical components of the 6 batches of Plantago asiatica and 7 batches of Plantago asiatica medicinal materials were stable between batches, while the similarities of two batches of mixed batches of original Plantago asiatica medicinal materials were 0.867 and 0.859, which were <0.9, indicating that there were certain differences between the batches of mixed batches of original Plantago asiatica, which may be related to the source ratio of the medicinal materials.

[0207] 2.2 Identification methods of different parts of Chinese medicinal materials of Plantago asiatica and different parts of Chinese medicinal materials of Plantago asiatica

[0208] 2.2.1 Variance analysis of peak areas of different parts of Plantago asiatica

[0209] The peak areas of the chromatograms of 24 samples of the whole herb, leaves, rhizomes and spikes of Plantago asiatica from batches P1 to P6 under item "2.1.5" were processed and imported into SPSS25.0 for single-factor ANOVA variance analysis. According to the results of the variance homogeneity test, most of the chromatographic peak areas had homogeneous variances. The peak areas of these chromatographic peaks with homogeneous variances were subjected to Bonferroni multiple comparison significance analysis, and a few with unequal variances were subjected to non-parametric multiple comparison analysis using Tamhane T2. The results of variance analysis of fingerprint peak areas of 6 batches of Plantago asiatica from different parts (unit: mAU*min) are shown in Table 2. It can be seen from Table 2 that peaks 4, 5, 11 (plantain glycosides), 16 and 18 (plantaginoside D) in the 6 batches of Plantago asiatica are mainly derived from rhizomes, peaks 1 (geniposide), 3, 7, 8, 9, 22, 23, 25 (apigenin), 26, 27 and 29 are mainly derived from spikes, peaks 2 (1-caffeoylquinic acid), 10, 13 (luteolin), 14, 15, 17 (verbascoside), 19 (isoverbascoside), 20 (apigenin-7-O-glucuronide), 21, 24 (luteolin) and 28 are mainly derived from leaves, and peaks 6 and 12 are basically not detected in various parts of Plantago asiatica. The significance results of the peak areas of each part showed that the differences in the peak areas of the following chromatographic peaks in different parts were statistically significant (P < 0.05), among which peak 1 ear > stem, peak 2 leaf, whole herb > stem > ear, peak 3 ear > stem, peak 7 ear > whole herb > stem > leaf, peak 8 ear > whole herb > stem, peak 9 ear > stem, peak 14 leaf, whole herb > ear > stem, peak 23 ear > stem.

[0210] Table 2

[0211]

[0212]

[0213] Note: The significance among different parts is represented by a for whole plant, b for stem, c for spike, and d for leaf

[0214] 2.2.2 Analysis of variance of peak areas of different parts of Plantago

[0215] The peak areas of the chromatograms of 28 samples of the whole herb, leaves, rhizomes and spikes of Plantago from batches C1 to C7 under item "2.1.5" were processed and imported into SPSS25.0 for single-factor ANOVA analysis of variance. According to the results of the variance homogeneity test, most of the chromatographic peak areas had homogeneous variances. The peak areas of these chromatographic peaks with homogeneous variances were subjected to Bonferroni multiple comparison significance analysis, and the peak areas of a few chromatographic peaks with unequal variances were subjected to non-parametric multiple comparison analysis using Tamhane T2. The results of variance analysis of the peak areas of fingerprint spectra of different parts of 7 batches of Plantago (unit: mAU*min) are shown in Table 3. It can be seen from Table 3 that peak 7 and peak 13 (luteolin) in the 7 batches of Plantago medicinal materials are mainly derived from the stem part, peak 1 (geniposide), peak 8, peak 17 (verbascoside), peak 22, peak 26, and peak 29 are mainly derived from the panicle part, and peak 2 (1-caffeoylquinic acid), peak 3, peak 4, peak 5, peak 6, peak 9, peak 10, peak 11 (plantainoside), peak 12, peak 14, peak 15, peak 16, peak 18 (plantaginoside D), peak 19 (isoverbascoside), peak 20 (apigenin-7-O-glucuronide), peak 21, peak 23, peak 24 (luteolin), peak 25 (apigenin), peak 27, and peak 28 are mainly derived from the leaf part. The significance results of the peak areas of each part showed that the differences in the peak areas of the following chromatographic peaks in different parts were statistically significant (P < 0.05), among which Peak 2 leaf > ear > stem, Peak 3 leaf > stem (undetectable in stem), Peak 5 and Peak 6 leaf, ear, whole herb > stem, Peak 7 leaf, whole herb > stem, Peak 11 and Peak 16 leaf > ear, stem, Peak 17 ear > leaf > stem, Peak 23 ear > stem.

[0216] Table 3

[0217]

[0218]

[0219] Note: The significance among different parts is represented by a for whole plant, b for stem, c for spike, and d for leaf

[0220] 2.2.3 Independent sample t-test of peak areas of Plantago asiatica and different parts of Plantago asiatica

[0221] The peak areas of the chromatograms of 24 samples of the whole grass, leaves, rhizomes, and spikes of P. plantaineum from batches P1 to P6 under item "2.1.5" and the peak areas of the chromatograms of 28 samples of the whole grass, leaves, rhizomes, and spikes of P. plantaineum from batches C1 to C7 under item "2.1.5" were processed and imported into SPSS25.0 for independent sample t-tests. After normality test, the significant results were calculated. The results of the independent sample t-test (unit: mAU*min) of the peak areas of different parts of P. plantaineum and P. plantaineum are shown in Table 4. As can be seen from Table 4, there is no statistical difference in the peak area of ​​peak 1 (geniposide acid) between P. plantaineum and P. plantaineum (P>0.05); the peak area of ​​peak 2 (1-caffeoylquinic acid) in the stems and spikes of P. plantaineum is lower than that in P. plantaineum (P<0.05); the peak area of ​​peak 3 in the whole grass, stems, spikes, and leaves of P. plantaineum is higher than that in P. plantaineum (P<0 .05); the peak area of ​​peak 4 in the ear of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 5 in the whole plant, stem, ear and leaf of Plantago asiatica was lower than that in Plantago asiatica (P<0.05); the peak areas of peak 7 and peak 8 in the whole plant, stem, ear and leaf of Plantago asiatica were higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 9 in the whole plant, stem and leaf of Plantago asiatica was lower than that in Plantago asiatica (P<0.05); the peak area of ​​peak 11 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 12 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 13 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 14 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 15 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 16 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0.05); the peak area of ​​peak 17 (plantain glycoside) in the whole plant, stem and leaf of Plantago asiatica was higher than that in Plantago asiatica (P<0 The peak areas of peak 14 in the whole plant, stem and leaves of Plantago asiatica were higher than those of Plantago asiatica (P<0.05); the peak areas of peak 15 in the whole plant and stem of Plantago asiatica were higher than those of Plantago asiatica (P<0.05); the peak areas of peak 16 and peak 18 (phytoside D) in the whole plant, ear and stem of Plantago asiatica were lower than those of Plantago asiatica (P<0.05); the peak areas of peak 17 (verbascoside) in the whole plant, stem and leaves of Plantago asiatica were higher than those of Plantago asiatica (P<0.05). The peak areas of peak 19 (isoverbascoside) in the stem, ear and leaf of Plantago asiatica were higher than those in Plantago asiatica (P<0.05); the peak areas of peak 21 in the stem and leaf of Plantago asiatica were lower than those in Plantago asiatica (P<0.05); the peak areas of peak 22 in the whole plant, ear and leaf of Plantago asiatica were higher than those in Plantago asiatica (P<0.05); the peak areas of peak 23 and peak 29 in the whole plant, stem, ear and leaf of Plantago asiatica were higher than those in Plantago asiatica (P<0.05). Among them, peak 6 and peak 12 were basically undetectable in Plantago asiatica and mainly existed in Plantago asiatica.

[0222] Table 4

[0223]

[0224]

[0225]

[0226] Note: The significance between different parts of Plantago and Plantago asiatica is marked with "*", where "*" indicates significance P<0.01, and "**" indicates significance P<0.001

[0227] 2.2.4 Identification criteria

[0228] Combined with the fingerprints of different parts of the Chinese medicinal materials of Plantago asiatica, combined with the variance analysis and independent sample t-test of the peak areas of different parts of the Chinese medicinal materials of Plantago asiatica, combined with the relative peak areas of different parts of the Chinese medicinal materials of Plantago asiatica, it is concluded that:

[0229] The identification criteria for different parts of the Chinese medicinal material Plantago asiatica can be:

[0230] The characteristic peak of verbascoside was recorded as peak 17, which was used as a reference peak. The characteristic peak with a retention time ratio of 0.943 to 0.950 to peak 17 was recorded as peak 16, and the characteristic peak with a retention time ratio of 1.350 to 1.360 to peak 17 was recorded as peak 21.

[0231] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica shows a characteristic peak with a retention time consistent with the fingerprint of the rhizome, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.006-0.029; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0-0.009, then the part to be tested is determined to be the rhizome;

[0232] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.001 to 0.004; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.002 to 0.011, then the part to be tested is determined to be the ear;

[0233] If the chromatogram of the tested part of the Chinese medicinal material Plantago asiatica shows a characteristic peak with a retention time consistent with the above-mentioned fingerprint spectrum of the leaves, and meets the following conditions at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.002-0.005; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.012-0.024, then the tested part is determined to be the leaf.

[0234] Combined with the fingerprints of different parts of Plantago Chinese medicinal materials, combined with the variance analysis and independent sample t-test of the peak areas of different parts of the above-mentioned Plantago Chinese medicinal materials, combined with the relative peak areas of different parts of the Plantago Chinese medicinal materials, it is concluded that:

[0235] The identification criteria for different parts of Plantago asiatica can be:

[0236] The characteristic peak of plantagoside D is recorded as peak 18, and with it as the reference peak, the characteristic peak with a retention time ratio of 0.944 to 0.966 to peak 18 is recorded as peak 17 (verbascoside), and the characteristic peak with a retention time ratio of 1.272 to 1.286 to peak 18 is recorded as peak 21;

[0237] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the rhizome, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0-0.392; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.226-0.655, then the part to be tested is determined to be the rhizome;

[0238] If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 1.119-3.312; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.228-1.076, then the part to be tested is determined to be the ear;

[0239] If the chromatogram of the tested part of the Plantago Chinese medicinal material shows a characteristic peak with a retention time consistent with the above-mentioned fingerprint spectrum of the leaves, and at the same time meets the following conditions: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0.091~0.671; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.676~5.169, then the tested part is determined to be the leaf.

[0240] 2.3 Identification methods of Chinese medicinal materials of Plantago asiatica, Chinese medicinal materials of Plantago asiatica and mixed Chinese medicinal materials of Plantago asiatica

[0241] 2.3.1 Variance analysis of peak area of ​​Plantago asiatica, Plantago asiatica and mixed-base Plantago asiatica

[0242] The peak areas of the chromatograms of the six samples of the whole herb of Plantago asiatica from batches P1 to P6, the peak areas of the chromatograms of the seven samples of the whole herb of Plantago asiatica from batches C1 to C7, and the peak areas of the chromatograms of the eight samples of the whole herb of Plantago asiatica from batches H1 to H8 under item "2.1.5" were processed and imported into SPSS25.0 for single-factor ANOVA analysis of variance. According to the results of the variance homogeneity test, most of the chromatographic peak areas had homogeneity of variance. The peak areas of the chromatographic peaks with homogeneity of variance were subjected to Bonferroni multiple comparison significance analysis, and the peak areas of the chromatographic peaks with unequal variance were subjected to non-parametric multiple comparison analysis using Tamhane T2. The results of fingerprint peak area variance analysis (unit: mAU*min) of 6 batches of whole herbs of Plantago asiatica, 7 batches of whole herbs of Plantago asiatica and 8 mixed-origin whole herbs of Plantago asiatica are shown in Table 5. It can be seen from Table 5 that the herbal medicines of Plantago asiatica from different sources are sorted according to the chromatographic peaks with statistically significant differences in peak area values, and are mainly divided into the following four categories: the first category, peak 5, peak 9, peak 11 (macroside), peak 16, Plantago > mixed-origin Plantago > Plantago asiatica; the second category, peak 7, peak 14, peak 17 (verbascoside), peak 19 (isoverbascoside), peak 22, peak 23, peak 27, Plantago asiatica > mixed batch > Plantago asiatica; the third category, peak 6, peak 12, Plantago > mixed batch > Plantago asiatica (not detected); the fourth category, peak 8, Plantago asiatica > mixed batch > Plantago asiatica (not detected). In addition, peak 3 is a significant difference peak between Plantago asiatica and Plantago asiatica. Therefore, the above 15 peaks can be used as important identification characteristic peaks of Plantago asiatica, Plantago asiatica and mixed batch medicinal materials.

[0243] Table 5

[0244]

[0245] Note: The significance of different medicinal materials is represented by e for Plantago asiatica, f for Plantago asiatica, and g for mixed batches.

[0246] 2.3.2 Study on fingerprint chemical pattern recognition of Chinese medicinal materials of Plantago asiatica, Chinese medicinal materials of Plantago asiatica and mixed Chinese medicinal materials of Plantago asiatica

[0247] 2.3.2.1 Principal component analysis (PCA) of plain plantain, plantain and mixed batch original plantain

[0248] The peak area data of the chromatograms of the six samples of the whole plant of P1 to P6 batches, the seven samples of the whole plant of C1 to C7 batches, and the eight samples of the whole plant of mixed origin from H1 to H8 batches were standardized and imported into SIMCA14.0 for principal component analysis. The principal component score diagram obtained is as follows: Fig.14As shown, 5 principal components were extracted with eigenvalues ​​> 1, and the cumulative variance contribution rate was 84.9%, which can reflect the overall information of 21 batches of plantain.

[0249] 2.3.2.2 Cluster analysis of plain plantain, plantain and mixed batch original plantain (HCA)

[0250] The cluster analysis diagram of 6 samples of whole herb of Plantago asiatica from batches P1 to P6, 7 samples of whole herb of Plantago asiatica from batches C1 to C7, and 8 samples of whole herb of mixed origin Plantago asiatica from batches H1 to H8 under item "2.1.5" is shown in the figure below: Fig.15 As shown, 6 batches of plantain are obviously clustered into one category, 4 batches of plantain are obviously clustered into one category, and the remaining plantains are clustered into one category with the mixed batch of plantain herbs. It can be seen that the overall quality of the mixed batch of plantain herbs is similar to that of plantain.

[0251] 2.3.2.3 Partial least squares regression analysis (PLS-DA) of plain plantain, plantain and mixed batch original plantain

[0252] The peak areas of the chromatograms of the six samples of the whole herb of Plantago asiatica from batches P1 to P6, the seven samples of the whole herb of Plantago asiatica from batches C1 to C7, and the eight samples of the whole herb of Plantago asiatica from batches H1 to H8 under item "2.1.5" were processed and respectively imported into SIMCA for PLS-DA analysis. The 21 batches of samples were subjectively classified according to the known categories, see Fig.16 As can be seen from the figure, the samples can be clearly divided into three categories. Among them, R2Y reaches 0.924, indicating that the model has a good fit, and Q2 reaches 0.72, indicating that the model has a strong predictive ability. The projection importance (VIP) of the 29 variables in the model is further analyzed, and the results are shown in Fig.17 Variables with VIP values ​​greater than 1 were selected as important quality indicators for the classification of plantain, plantain and mixed batch plantain samples. A total of 15 important characteristic indicators were extracted, which were consistent with the results of variance analysis of the three. The importance ranking was peak 3, peak 8, peak 28, peak 12, peak 14, peak 7, peak 5, peak 17, peak 6, peak 19, peak 23, peak 11, peak 22, peak 27, peak 9, and peak 16.

[0253] 2.3.3 Identification criteria

[0254] Combining the fingerprints of Plantago asiatica, Plantago asiatica and mixed-based original plantain Chinese medicinal materials, combined with the variance analysis and chemical pattern recognition research of the peak areas of the above-mentioned Plantago asiatica, Plantago asiatica and mixed-based original plantain Chinese medicinal materials, combined with the relative peak areas of Plantago asiatica, Plantago asiatica and mixed-based original plantain Chinese medicinal materials, it is concluded that:

[0255] The identification criteria of plantain herbs from different sources can be:

[0256] The characteristic peak of plantagoside is recorded as peak 11, and the characteristic peak with a retention time ratio of 1.346 to 1.372 with respect to peak 11 is recorded as peak 17 (verbascoside);

[0257] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the whole herb of the Chinese medicinal material Plantago asiatica, and the ratio of the peak area of ​​peak 17 to peak 11 is 48.799-152.931, it is determined that the Chinese medicinal material to be tested is the whole herb of the Chinese medicinal material Plantago asiatica;

[0258] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the whole herb of the Chinese medicinal material Plantago, and the ratio of the peak area of ​​peak 17 to peak 11 is 0.013-0.055, then it is determined that the Chinese medicinal material to be tested is the whole herb of the Chinese medicinal material Plantago;

[0259] If the chromatogram of the Chinese medicinal material to be tested shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the above-mentioned mixed-base original plantain Chinese medicinal material, and the ratio of the peak area of ​​peak 17 to peak 11 is 0.148-2.326, then it is determined that the Chinese medicinal material to be tested is a mixed-base original plantain Chinese medicinal material.

[0260] 2.4 Quality evaluation methods for Chinese medicinal materials of Plantago asiatica, Chinese medicinal materials of Plantago asiatica and mixed Chinese medicinal materials of Plantago asiatica

[0261] 2.4.1 CRITIC method

[0262] The CRITIC method is an objective weighting method that calculates weights based on the importance criterion of inter-layer correlation. It is applicable to data with correlation between the analyzed indicators. It combines the impact of data volatility and data correlation on weights, and determines indicator weights based on volatility (data standard deviation) and conflict between indicators. Among them, volatility takes data standard deviation, and conflict between indicators is explained by correlation between indicators. If there is a strong positive correlation between two indicators, it means that the conflict between the two indicators is low. The weighting steps are as follows:

[0263] 2.4.1.1 Data Homogenization

[0264] Obtain the peak area values ​​x of 29 chromatographic peaks (representing n evaluation indicators) in 21 batches (representing m evaluation objects) of Plantain from batches P1 to P6, batches C1 to C7, and batches H1 to H8 under item "2.1.5" ij , where i represents the batch of plantain (evaluation object), i = 1, 2, 3...m, and j represents the chromatographic peak (evaluation index), j = 1, 2, 3...n. According to formula (1), the numerical matrix R is obtained: ij, where R represents a numerical matrix. The 29 chromatographic peaks used in this study are positive indicators R ij , positive indicators are also called benefit indicators, the larger the indicator value, the better; negative indicators are also called cost indicators, the smaller the indicator value, the better. The negative indicator is obtained according to formula (2) The numerical matrix R ij , which is not involved in this study.

[0265] Positive indicators

[0266]

[0267] Negative indicators

[0268]

[0269] 2.5.1.2 Calculating volatility and conflict

[0270] Logarithmic matrix R ij Calculate the standard deviation sd and get the volatility S j .

[0271] The peak area value x ij Imported into SPSS25.0, the correlation matrix (correlation coefficient is Pearson correlation coefficient r ij ), and the conflict A is obtained by formula (3): j The data are shown in Table 6.

[0272]

[0273] 2.5.1.3 Calculating Information and Weight

[0274] The information volume C after the two are associated is obtained by formula (4): j .

[0275] The weight coefficients W of 29 indicators of 21 batches of plantain herbs were obtained by formula (5): j The data are shown in Table 6.

[0276] C j =S j ×A j (4)

[0277]

[0278] Table 6

[0279]

[0280]

[0281] 2.5.2 Comprehensive evaluation of medicinal material quality based on grey correlation degree of CRITIC method

[0282] 2.5.2.1 Optimal sequence selection

[0283] Since the dimensions of the evaluation indicators (29 chromatographic peaks) of the 21 batches of plantain are inconsistent, the peak area values ​​of the original characteristic peaks of the evaluation indicators are averaged to obtain the dimensionless unit sequence x' i’ (j), see formula (6). The maximum unit sequence of the 29 indicators after dimensionless transformation is used as the optimal reference sequence x0(j).

[0284]

[0285] 2.5.2.2 Calculation of correlation coefficient

[0286] Refer to formula (7) to calculate the correlation coefficient, and the results are shown in Table 7. Where ρ is the resolution coefficient. The smaller ρ is, the greater the resolution is. Generally, the value range of ρ is [0, 1]. When ρ≤0.5463, the resolution is the best, and ρ=0.5 is usually taken.

[0287] Referring to formula (8), the correlation is calculated. It can be seen that the correlations of the 29 index peaks are 0.41, 0.68, 0.63, 0.73, 0.74, 0.72, 0.61, 0.47, 0.51, 0.46, 0.24, 0.34, 0.49, 0.44, 0.39, 0.58, 0.24, 0.58, 0.43, 0.49, 0.57, 0.46, 0.47, 0.43, 0.55, 0.48, 0.65, 0.51, and 0.64.

[0288]

[0289]

[0290] Table 7 (%)

[0291]

[0292]

[0293] 2.5.2.3 Comprehensive evaluation of grey correlation based on CRITIC method

[0294] The weight coefficient W of each indicator calculated by the CRITIC method j As shown in Table 6, combined with the weight coefficient W j According to formula (8), the comprehensive evaluation results of the grey correlation degree of 21 batches of plantain based on the CRITIC method were obtained. i, and the quality ranking is carried out accordingly, and the results are shown in Table 8. From the data in Table 8, it can be seen that the top 5 in quality ranking are H6, C3, C2, P2, and C6, and the quality is mainly concentrated in the plantago, the comprehensive quality evaluation of mixed batches, flat plantago, and plantago i The average values ​​were 2.40%, 2.18%, and 2.46%, respectively, indicating that the overall quality of Plantain was the best, followed by mixed batch Plantain, and Plantain was the worst. i The larger the value, the better the quality of the Chinese medicinal materials to be tested.

[0295] Table 8

[0296]

[0297]

[0298]

[0299] The present invention constructs fingerprints of different origins of plantain medicinal materials, which can be used to authenticate the source composition of plantain medicinal materials according to the characteristics that the ingredients contained in different sources of plantain are different.

[0300] The present invention performs comparative analysis of variance analysis and independent sample T test, as well as chemical identification pattern (PCA, HCA and PLS-DA) analysis, and respectively screens out identification indicators and importance rankings of respective identification models for identification of different parts of Plantago asiatica, identification of different parts of Plantago asiatica, and identification of Plantago asiatica, Plantago asiatica and mixed-based original Plantain, and can be used for analytical identification work in these situations.

[0301] The present invention adopts the CRITIC method to objectively weight each index component in the fingerprint spectrum, determines the index weight value of each analyzed sample, and combines the grey correlation degree to perform the overall quality evaluation of the three sources of plantain herbs. The quality method based on the CRITIC method and the grey correlation degree can be used to rank and evaluate the quality differences of multiple batches of plantain herbs from different sources.

[0302] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0303] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for constructing fingerprints of Chinese medicinal materials including Plantago asiatica, Plantago asiatica and mixed-origin Plantago asiatica, characterized in that: The following steps are involved: The whole plant and predetermined parts of the Chinese medicinal material Plantago asiatica are taken, crushed, added with an extraction solvent, subjected to ultrasonic treatment, and the extract is collected to prepare a Plantago asiatica test solution, wherein the predetermined parts are rhizomes, leaves, and spikes; The whole plant and predetermined parts of the Chinese medicinal material Plantago are taken, crushed, added with an extraction solvent, and ultrasonically treated, and the extract is collected to prepare a Plantago test solution, wherein the predetermined parts are rhizomes, leaves, and spikes; Take the mixed-origin plantain Chinese medicinal material, crush it, add the extraction solvent, perform ultrasonic treatment, collect the extract, and prepare the mixed-origin test solution; Taking the plantain test solution, the plantago test solution and the mixed base test solution for ultra-high performance liquid chromatography determination; The chromatographic conditions of the ultra-high performance liquid chromatography include: The chromatographic column was an Aglient SB C18 column, 2.1 × 100 mm, 1.6 μm; The mobile phase includes a mobile phase A and a mobile phase B, wherein the mobile phase A is acetonitrile, and the mobile phase B is a phosphoric acid aqueous solution, wherein the volume fraction of phosphoric acid in the phosphoric acid aqueous solution is 0.1%, and the gradient elution is performed at 0-3 min, the detection wavelength is 245 nm, 3 min-48 min, the detection wavelength is 330 nm, and 48 min-60 min, the detection wavelength is 245 nm; The gradient elution includes the following procedures: From 0 to 2 min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%; From 2 min to 4 min, the volume fraction of the mobile phase A increased from 10% to 13%, and the volume fraction of the mobile phase B decreased from 90% to 87%; From 4 min to 14 min, the volume fraction of the mobile phase A was maintained at 13%, and the volume fraction of the mobile phase B was maintained at 87%; From 14 min to 24 min, the volume fraction of the mobile phase A increased from 13% to 15%, and the volume fraction of the mobile phase B decreased from 87% to 85%; From 24 to 25 minutes, the volume fraction of the mobile phase A increased from 15% to 17%, and the volume fraction of the mobile phase B decreased from 85% to 83%; From 25 min to 31 min, the volume fraction of the mobile phase A increased from 17% to 22%, and the volume fraction of the mobile phase B decreased from 83% to 78%; From 31min to 41min, the volume fraction of the mobile phase A increased from 22% to 40%, and the volume fraction of the mobile phase B decreased from 78% to 60%; From 41min to 50min, the volume fraction of the mobile phase A increased from 40% to 68%, and the volume fraction of the mobile phase B decreased from 60% to 32%; From 50min to 53min, the volume fraction of the mobile phase A increased from 68% to 88%, and the volume fraction of the mobile phase B decreased from 32% to 12%; From 53min to 54min, the volume fraction of the mobile phase A decreased from 88% to 10%, and the volume fraction of the mobile phase B increased from 12% to 90%; From 54 min to 60 min, the volume fraction of the mobile phase A was maintained at 10%, and the volume fraction of the mobile phase B was maintained at 90%.

2. The construction method according to claim 1, characterized in that: The chromatographic conditions of the ultra-high performance liquid chromatography also include at least one of the following conditions: Column temperature is 28°C~32°C; Flow rate: 0.25mL / min~0.35mL / min; The injection volume is 1μL~2μL.

3. The construction method according to claim 1 or 2, characterized in that: Also includes at least one of the following features: The extraction solvent for preparing the Plantago asiatica test solution is a methanol-water solution; For every 1g of the whole plant and the predetermined parts of the Chinese medicinal material Plantago asiatica, 40mL to 60mL of the extraction solvent is added.

4. The construction method according to claim 3, characterized in that: Also includes at least one of the following features: The volume fraction of methanol in the methanol aqueous solution for preparing the plantain test solution is 40% to 80%; The ultrasonic treatment time for preparing the Plantago asiatica test solution is 20 min to 40 min.

5. The construction method according to claim 1, characterized in that: Also includes at least one of the following features: The extraction solvent for preparing the Plantago test solution is methanol-water solution; For every 1g of the whole plant and the predetermined parts of the Chinese medicinal material Plantago, 40mL to 60mL of the extraction solvent is added.

6. The construction method according to claim 5, characterized in that: Also includes at least one of the following features: The volume fraction of methanol in the methanol aqueous solution for preparing the Plantago test solution is 40% to 80%; The ultrasonic treatment time for preparing the Plantago test solution is 20 min to 40 min.

7. The construction method according to claim 1, characterized in that: Also includes at least one of the following features: The extraction solvent for preparing the mixed base original test solution is methanol-water solution; For every 1g of the mixed-base original plantain Chinese medicinal material, 40mL~60mL of the extraction solvent is added.

8. The construction method according to claim 7, characterized in that: Also includes at least one of the following features: The volume fraction of methanol in the methanol aqueous solution for preparing the mixed base original test solution is 40% to 80%; The ultrasonic treatment time for preparing the mixed base sample solution is 20min~40min.

9. A method for identifying different parts of a Chinese medicinal material Plantago asiatica, characterized in that: The following steps are involved: Taking the part of the Chinese medicinal material Plantago asiatica to be tested, crushing it, adding an extraction solvent to perform ultrasonic treatment, collecting the extract, and preparing a sample solution to be tested; According to the chromatographic conditions described in any one of claims 1 to 8, the sample solution to be tested is subjected to ultra-high performance liquid chromatography, the obtained chromatogram is matched with the fingerprint spectrum of the preset part of the Plantago asiatica Chinese medicinal material described in any one of claims 1 to 8, the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded, and the attribution of the part to be tested is determined.

10. The method for identifying different parts of the Chinese medicinal material Plantago asiatica according to claim 9, characterized in that: The identification criteria include: The characteristic peak of verbascoside was recorded as peak 17, which was used as a reference peak. The characteristic peak with a retention time ratio of 0.943 to 0.950 to peak 17 was recorded as peak 16, and the characteristic peak with a retention time ratio of 1.350 to 1.360 to peak 17 was recorded as peak 21. If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint of the rhizome according to any one of claims 1 to 4, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.006 to 0.029; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0 to 0.009, then the part to be tested is determined to be the rhizome; If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear as described in any one of claims 1 to 4, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.001 to 0.004; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.002 to 0.011, then the part to be tested is determined to be the ear; If the chromatogram of the tested part of the Chinese medicinal material Plantago asiatica shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the leaves described in any one of claims 1 to 4, and meets the following conditions at the same time: a) the ratio of the peak area of ​​peak 16 to peak 17 is 0.002 to 0.005; b) the ratio of the peak area of ​​peak 21 to peak 17 is 0.012 to 0.024, then the tested part is determined to be a leaf.

11. The method for identifying different parts of the Chinese medicinal material Plantago asiatica according to claim 9 or 10, characterized in that: Also includes at least one of the following features: The extraction solvent is methanol-water solution; For every 1g of the tested part of the Plantago asiatica Chinese medicinal material, 40mL to 60mL of the extraction solvent is added.

12. The method for identifying different parts of the Chinese medicinal material Plantago asiatica according to claim 11, characterized in that: Also includes at least one of the following features: The volume fraction of methanol in the methanol aqueous solution is 40% to 80%; The ultrasonic treatment time is 20 min to 40 min.

13. A method for identifying different parts of a Chinese medicinal material, Plantago, characterized in that: The following steps are involved: Taking the part of the Chinese medicinal material Plantago asiatica to be tested, crushing it, adding an extraction solvent, performing ultrasound, collecting the extract, and preparing a sample solution to be tested; According to the chromatographic conditions described in any one of claims 1 to 8, the sample solution to be tested is subjected to ultra-high performance liquid chromatography, the obtained chromatogram is matched with the fingerprint spectrum of the preset part of the Plantago Chinese medicinal material described in any one of claims 1 to 8, the corresponding characteristic peaks and the peak areas of the characteristic peaks are recorded, and the part to be tested is determined.

14. The method for identifying different parts of the Chinese medicinal material Plantago asiatica according to claim 13, characterized in that: The identification criteria include: The characteristic peak of plantagoside D is recorded as peak 18, and with it as the reference peak, the characteristic peak with a retention time ratio of 0.944 to 0.966 to peak 18 is recorded as peak 17, and the characteristic peak with a retention time ratio of 1.272 to 1.286 to peak 18 is recorded as peak 21; If the chromatogram of the part to be tested of the Chinese medicinal material Plantago shows a characteristic peak with a retention time consistent with the fingerprint of the rhizome described in any one of claims 5 to 6, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0 to 0.392; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.226 to 0.655, then the part to be tested is determined to be the rhizome; If the chromatogram of the part to be tested of the Chinese medicinal material Plantago asiatica has a characteristic peak with a retention time consistent with the fingerprint spectrum of the ear as described in any one of claims 5 to 6, and the following conditions are met at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 1.119 to 3.312; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.228 to 1.076, then the part to be tested is determined to be the ear; If the chromatogram of the tested part of the Plantago Chinese medicinal material shows a characteristic peak with a retention time consistent with the fingerprint spectrum of the leaf described in any one of claims 5 to 6, and satisfies the following conditions at the same time: a) the ratio of the peak area of ​​peak 17 to peak 18 is 0.091~0.671; b) the ratio of the peak area of ​​peak 21 to peak 18 is 0.676~5.169, then the tested part is determined to be a leaf.

15. The method for identifying different parts of the Chinese medicinal material Plantago according to claim 13 or 14, characterized in that: Also includes at least one of the following features: The extraction solvent is methanol-water solution; For every 1g of the tested part of the Plantago Chinese medicinal material, 40mL~60mL of the extraction solvent is added.

16. The method for identifying different parts of the Chinese medicinal material Plantago asiatica according to claim 15, characterized in that: Also includes at least one of the following features: The volume fraction of methanol in the methanol aqueous solution is 40% to 80%; The ultrasonic treatment time is 20 min to 40 min.

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