Fingerprint construction method and identification method of psoralea corylifolia and salt psoralea corylifolia Chinese medicine formula granules

Abstract

The present application relates to the technical field of Chinese medicine detection and analytical chemistry, and specifically to a fingerprint construction method and identification method of Chinese medicine formula granules of Psoralea corylifolia and Psoralea corylifolia salt. The fingerprint construction method of the present application includes the use of ultra-high performance liquid chromatography for analysis, and the chromatographic conditions are methanol as mobile phase A and a formic acid aqueous solution with a volume concentration of 0.08% to 0.12% as mobile phase B. The fingerprint of the present application is characteristic, reproducible and operable, and can provide a scientific basis for the quality evaluation and control of the above-mentioned Chinese medicine formula granules. The present application also relates to an identification method of Chinese medicine formula granules based on the above-mentioned fingerprint, which can effectively identify Chinese medicine formula granules of Psoralea corylifolia and Psoralea corylifolia salt.

Description

Technical Field

[0001] The present application relates to the technical field of traditional Chinese medicine detection and analytical chemistry, and specifically to a fingerprint spectrum construction method and an identification method of psoralea corylifolia and psoralea corylifolia salt traditional Chinese medicine formula granules. Background Art

[0002] Chinese medicine formula granules are granules made from single Chinese medicinal materials (cooked pieces) through extraction, concentration, drying, granulation and other processes. Compared with cooked pieces, it is difficult to accurately identify and distinguish them from their appearance. In order to ensure the accuracy and safety of clinical medication, it is necessary to establish corresponding detection methods for different formula granules.

[0003] Psoralea corylifolia L is a dried mature fruit of the leguminous plant Psoralea corylifolia L. Its surface is black, dark brown or grayish brown with fine reticular wrinkles. It has the effects of warming the kidney and yang, relieving asthma and warming the spleen and stopping diarrhea. Psoralea corylifolia raw product has certain toxic side effects and is generally used after processing. The commonly used processed product in clinical practice is salt psoralea corylifolia slices. Its properties, processing methods and active ingredient content have been clearly defined in the pharmacopoeia, but there are few reports on its formula granules.

[0004] There is a method involving an infrared spectroscopy identification method for psoralea corylifolia and salt psoralea corylifolia formula granules. This method has certain limitations and has not been well promoted and applied. Summary of the invention

[0005] In view of this, the present application proposes a method for constructing a fingerprint spectrum of Psoralea corylifolia and Psoralea corylifolia salt Chinese medicine formula granules, which can objectively, scientifically and comprehensively reflect the intrinsic chemical characteristic information of the Chinese medicine formula granules as a basis for their quality detection.

[0006] Specifically, the method for constructing the fingerprint spectrum of the traditional Chinese medicine formula granules comprises the following steps:

[0007] Accurately weigh the Chinese medicine formula granules, extract them with an extraction solvent, and prepare a test solution;

[0008] Accurately weigh the standard products of psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol, and dissolve them in a solvent to prepare a reference solution;

[0009] Accurately aspirate the test sample solution and the reference solution, and perform ultra-high performance liquid chromatography analysis respectively to obtain a test sample fingerprint and a reference chromatogram, and obtain a fingerprint of the Chinese medicine formula granules according to the test sample fingerprint and the reference chromatogram;

[0010] The ultra-high performance liquid chromatography analysis includes elution according to the following mobile phase conditions: methanol is used as mobile phase A, and a formic acid aqueous solution with a volume concentration of 0.08% to 0.12% is used as mobile phase B, and the total volume percentage of the mobile phase A and the mobile phase B is 100%.

[0011] In some embodiments of the present application, in the fingerprint construction method, the elution procedure includes:

[0012] From 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%;

[0013] From 9min to 18min, the volume percentage of the mobile phase A increases from 30% to 46%;

[0014] From 18min to 19min, the volume percentage of the mobile phase A increased from 46% to 57%;

[0015] From 19min to 34min, the volume percentage of the mobile phase A increased from 57% to 77%;

[0016] From 34min to 42min, the volume percentage of the mobile phase A increases from 77% to 90%;

[0017] 42min~42.1min, the volume percentage of the mobile phase A decreases from 90% to 18%;

[0018] From 42.1 min to 47 min, the volume percentage of the mobile phase A was maintained at 18%.

[0019] In some embodiments of the present application, in the fingerprint construction method, the conditions of the ultra-high performance liquid chromatography analysis include one or more of the following characteristics:

[0020] (1) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0021] (2) The model of the chromatographic column is ACQUITY UPLC BEH;

[0022] (3) Column temperature is 38°C to 42°C;

[0023] (4) The detection wavelength is 240nm to 250nm;

[0024] (5) The flow rate of the mobile phase is 0.28 mL / min to 0.32 mL / min.

[0025] In some embodiments of the present application, in the fingerprint construction method, the extraction solvent is methanol, and the extraction method is ultrasound.

[0026] In some embodiments of the present application, in the fingerprint spectrum construction method, the psoralea corylifolia Chinese medicine formula granules are prepared by comprising the following steps:

[0027] Take the psoralea corylifolia slices, add water in an amount of 6 to 16 times the mass of the psoralea corylifolia slices, heat and boil for 0.5 to 2.5 hours; continue to add water in an amount of 5 to 12 times the mass of the psoralea corylifolia slices, heat and boil for 0.5 to 2.0 hours, filter, decompress the obtained filtrate to a clear paste with a relative density of 1.02 to 1.10, filter, dry, and granulate.

[0028] In some embodiments of the present application, in the fingerprint spectrum construction method, the salt psoralea corylifolia Chinese medicine formula granules are prepared by comprising the following steps:

[0029] Take salt psoralea corylifolia slices, add water in an amount of 6 to 16 times the mass of the salt psoralea corylifolia slices, heat and boil for 0.5 to 2.5 hours; continue to add water in an amount of 5 to 12 times the mass of the salt psoralea corylifolia slices, heat and boil for 0.5 to 2.0 hours, filter, and concentrate the obtained filtrate under reduced pressure to a clear paste with a relative density of 1.02 to 1.10, filter, dry, and granulate.

[0030] The present application also relates to a method for identifying Chinese medicine formula granules, comprising the following steps:

[0031] According to the fingerprint construction method, the fingerprint of the Psoralea corylifolia Chinese medicine formula granules and the fingerprint of the Psoralea corylifolia salt Chinese medicine formula granules are obtained;

[0032] The fingerprint of the Chinese medicine formula granules of Psoralea corylifolia and the fingerprint of the Chinese medicine formula granules of Psoralea corylifolia salt are compared, and a total of 22 common peaks are calibrated, and the ratio of the peak area and the sampling amount of each peak is used as the data for orthogonal partial least squares discriminant analysis, and 9 marker peaks are determined according to the analysis results;

[0033] SPSS20 software was used to perform discriminant analysis based on the 9 marker peaks, and a discriminant equation was established; wherein the retention times of the 9 marker peaks were as follows: peak 9: 11.88±10%, peak 15: 17.54±10%, peak 3: 6.46±10%, peak 12: 15.62±10%, peak 5: 7.89±10%, peak 14: 17.03±10%, peak 11: 14.79±10%, peak 13: 16.33±10% and peak 4: 7.01±10%;

[0034] Obtaining the centroid value I of the Psoralea corylifolia Chinese medicine formula granules and the centroid value II of the Psoralea corylifolia salt Chinese medicine formula granules;

[0035] Extracting the Chinese medicine formula granules to be tested with an extraction solvent to prepare a test product solution;

[0036] The test solution is subjected to ultra-high performance liquid chromatography to obtain the peak areas of the nine marker peaks, and the ratio of the peak areas of the nine marker peaks to the sampling amount is used as data, substituted into the discriminant equation, and the discriminant score is calculated;

[0037] If the discrimination score is close to the centroid value I, it is judged to be the Psoralea corylifolia Chinese medicine formula granules; if the discrimination score is close to the centroid value II, it is judged to be the Salt Psoralea corylifolia Chinese medicine formula granules;

[0038] The ultra-high performance liquid chromatography analysis includes elution according to the following mobile phase conditions: methanol is used as mobile phase A, and a formic acid aqueous solution with a volume concentration of 0.08% to 0.12% is used as mobile phase B, and the total volume percentage of the mobile phase A and the mobile phase B is 100%.

[0039] In some embodiments of the present application, in the identification method, the discriminant equation is: Y = -0.008×Peak 3-1.488×Peak 4+0.056×Peak 5+3.381×Peak 9-3.863×Peak 11-0.281×Peak 12+1.220×Peak 13+0.449×Peak 14-1.367×Peak 15-0.144.

[0040] In some embodiments of the present application, in the identification method, the elution procedure includes:

[0041] From 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%;

[0042] From 9min to 18min, the volume percentage of the mobile phase A increases from 30% to 46%;

[0043] From 18min to 19min, the volume percentage of the mobile phase A increased from 46% to 57%;

[0044] From 19min to 34min, the volume percentage of the mobile phase A increased from 57% to 77%;

[0045] From 34min to 42min, the volume percentage of the mobile phase A increases from 77% to 90%;

[0046] 42min~42.1min, the volume percentage of the mobile phase A decreases from 90% to 18%;

[0047] From 42.1 min to 47 min, the volume percentage of the mobile phase A was maintained at 18%.

[0048] In some embodiments of the present application, in the identification method, the conditions of the ultra-high performance liquid chromatography analysis include one or more of the following characteristics:

[0049] (1) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0050] (2) The model of the chromatographic column is ACQUITY UPLC BEH;

[0051] (3) Column temperature is 38°C to 42°C;

[0052] (4) The detection wavelength is 240nm to 250nm;

[0053] (5) The flow rate of the mobile phase is 0.28 mL / min to 0.32 mL / min.

[0054] In some embodiments of the present application, in the identification method, the extraction solvent is methanol, and the extraction method is ultrasound.

[0055] The fingerprint construction method of the present application adopts appropriate chromatographic conditions, and the obtained fingerprint is characteristic, reproducible and operable, and can provide more abundant characteristic peak information, thereby more comprehensively reflecting the quality of the sample. The method is stable, precise and reproducible, and provides a rapid, sensitive, objective and accurate detection method for the quality of Psoralea corylifolia Chinese medicine formula granules and Psoralea corylifolia salt Chinese medicine formula granules.

[0056] The method of this application is based on the interaction between specific Chinese medicine raw materials and chemical components in the preparation process. The retention time of each characteristic peak is fixed within a certain range and will be different when the raw materials change (such as the use of fake and inferior medicinal materials). Therefore, the quality of Chinese medicine formula granules can be well identified, which can play a role in distinguishing authenticity.

[0057] This application is based on the constructed fingerprint map (fingerprint map of Psoralea corylifolia and Psoralea corylifolia salt Chinese medicine formula granules), and 22 common peaks are established. Based on the above characteristic map construction, combined with the chemometric method, effective chemical information can be extracted to a greater extent, and the difference characteristic indicators can be screened out to establish a judgment function equation for pattern recognition, so as to effectively identify Psoralea corylifolia and Psoralea corylifolia salt Chinese medicine formula granules, making up for the defect that they cannot be inspected and identified from microscopic and property identification characteristics. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0059] Figure 1 This is a comparison chart of the extraction results under different methanol concentration conditions in 1.2.1 of this application;

[0060] Figure 2 This is a comparison chart of chromatographic results under different detection wavelength conditions in 1.3.1 of this application;

[0061] Figure 3 This is a comparison chart of the chromatographic results under different column temperature conditions in 1.3.2 of this application;

[0062] Figure 4 This is a comparison chart of the chromatographic results under different flow rate conditions in 1.3.3 of this application;

[0063] Figure 5 This is a comparison chart of the chromatographic results under different mobile phase conditions in 1.3.4 of this application;

[0064] Figure 6 This is a comparison chart of the chromatographic results under different gradient elution conditions in 1.3.5 of this application;

[0065] Figure 7 The fingerprints of 19 batches of Psoralea corylifolia Chinese medicine granules in Example 1;

[0066] Figure 8 The fingerprints of 19 batches of salt psoralea corylifolia Chinese medicine granules in Example 1;

[0067] Fig. 9 The chromatographic peak identification diagram of the Chinese medicine formula granules of Psoralea corylifolia in Example 1;

[0068] Fig.10 It is a score graph of the orthogonal partial least squares discriminant analysis of the Chinese medicine formula granules of Psoralea corylifolia and Psoralea corylifolia salt in Example 1;

[0069] Fig.11 This is a VIP value graph obtained by orthogonal partial least squares discriminant analysis of the Chinese medicine formula granules of Psoralea corylifolia and Psoralea corylifolia salt in Example 1. DETAILED DESCRIPTION

[0070] The present application will be further described below in conjunction with the embodiments, examples and accompanying drawings. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. In addition, it should be understood that after reading the content taught in the present application, those skilled in the art may make various changes or modifications to the present application, and these equivalent forms also fall within the scope of protection of the claims attached to the present application.

[0071] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0072] the term

[0073] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0074] Herein, "preferred", "preferable", "better", etc. are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of this application.

[0075] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0076] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0077] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0078] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±0.5°C, ±0.4°C, ±0.3°C, ±0.2°C, and ±0.1°C are allowed.

[0079] In the present application, weight may be μg, mg, g, kg or other mass units known in the chemical industry.

[0080] In the present application, unless otherwise specified, the size, particle size and diameter generally refer to the average value.

[0081] The present application relates to a fingerprint construction method of Chinese medicine formula granules of Psoralea corylifolia and Psoralea corylifolia salt. In the present application, the Chinese medicine formula granules of Psoralea corylifolia or Psoralea corylifolia salt are prepared by water extraction, specifically including the process of pretreatment, water extraction, concentration, drying and granulation.

[0082] For example, the Chinese medicine formula granules of Psoralea corylifolia can be prepared by the following steps: taking Psoralea corylifolia medicinal materials, selecting and removing impurities, crushing or breaking to obtain Psoralea corylifolia slices. Taking Psoralea corylifolia slices, adding water and decocting twice, adding water 6 to 16 times the amount of the raw material for the first time, heating and decocting for 0.5 to 2.5 hours; adding water 5 to 12 times the amount of the raw material for the second time, heating and decocting for 0.5 to 2.0 hours, filtering the decoction, and concentrating the filtrate under reduced pressure to a clear paste with a relative density of 1.02 to 1.10 (80°C), filtering, drying, and granulating.

[0083] For example, the salt psoralea corylifolia Chinese medicine formula granules can be prepared by the following steps: take clean psoralea corylifolia, stir-fry until slightly bulging according to the salt roasting method recorded in the "Chinese Pharmacopoeia General Principles 0213", crush or break, and obtain salt psoralea corylifolia slices. Take the salt psoralea corylifolia slices, add water and boil twice, add 6 to 16 times the amount of water for the first time, heat and boil for 0.5 to 2.5 hours; add 5 to 12 times the amount of water for the second time, heat and boil for 0.5 to 2.0 hours, filter the decoction, and reduce the pressure on the filtrate to a clear paste with a relative density of 1.02 to 1.10 (80°C), filter, dry, and granulate.

[0084] Compared with traditional psoralea corylifolia or salt psoralea corylifolia preparations, the Chinese medicine formula granules of the present application are based on water extraction, and the content of active ingredients such as psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol is higher.

[0085] Furthermore, the fingerprint spectrum construction method of the Chinese medicine formula granules of Psoralea corylifolia or Psoralea corylifolia salt comprises the following steps:

[0086] Accurately weigh the Chinese medicine formula granules, extract them with an extraction solvent, and prepare a test solution;

[0087] Accurately weigh the standard products of psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol, and dissolve them in a solvent to prepare a reference solution;

[0088] Accurately aspirate the test sample solution and the reference solution, and perform ultra-high performance liquid chromatography analysis respectively to obtain the test sample fingerprint spectrum I and the reference solution chromatogram, and obtain the fingerprint spectrum of the Chinese medicine formula granules according to the test sample fingerprint spectrum and the reference solution chromatogram;

[0089] The ultra-high performance liquid chromatography analysis includes elution according to the following mobile phase conditions: methanol is used as mobile phase A, and a formic acid aqueous solution with a volume concentration of 0.08% to 0.12% is used as mobile phase B, and the total volume percentage of the mobile phase A and the mobile phase B is 100%.

[0090] In some embodiments, in the fingerprint construction method, the elution procedure includes:

[0091] From 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%;

[0092] From 9min to 18min, the volume percentage of the mobile phase A increases from 30% to 46%;

[0093] From 18min to 19min, the volume percentage of the mobile phase A increased from 46% to 57%;

[0094] From 19min to 34min, the volume percentage of the mobile phase A increased from 57% to 77%;

[0095] From 34min to 42min, the volume percentage of the mobile phase A increases from 77% to 90%;

[0096] 42min~42.1min, the volume percentage of the mobile phase A decreases from 90% to 18%;

[0097] From 42.1 min to 47 min, the volume percentage of the mobile phase A was maintained at 18%.

[0098] In some embodiments, in the fingerprint construction method, the conditions of ultra-high performance liquid chromatography analysis include one or more of the following characteristics:

[0099] (1) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0100] (2) The model of the chromatographic column is ACQUITY UPLC BEH;

[0101] (3) Column temperature is 38°C to 42°C;

[0102] (4) The detection wavelength is 240nm to 250nm;

[0103] (5) The flow rate of the mobile phase is 0.28 mL / min to 0.32 mL / min.

[0104] In some embodiments, in the fingerprint construction method, the extraction solvent is methanol, which has a better extraction effect on fat-soluble components, can extract more chromatographic peaks, and more comprehensively characterize the sample. Furthermore, the extraction method is ultrasound.

[0105] The present application also relates to a method for identifying Chinese medicine formula granules, which can be used to identify the above-mentioned Psoralea corylifolia Chinese medicine formula granules and Psoralea corylifolia salt Chinese medicine formula granules.

[0106] Traditional identification technology is based on a small number of characteristic peaks, making it difficult to identify Psoralea corylifolia and Psoralea salt with similar active ingredients. The identification method of the present application has more characteristic peaks, and uses ultra-high performance liquid chromatography to construct fingerprints respectively under appropriate chromatographic conditions, with higher precision and accuracy. At the same time, it also combines chemometric methods to extract effective chemical information to a greater extent, screen out differential characteristic indicators to establish judgment function equations, perform pattern recognition, and effectively distinguish Psoralea corylifolia and Psoralea salt Chinese medicine formula granules.

[0107] In some embodiments, the method for identifying Chinese medicine granules comprises the following steps:

[0108] S100: Establishing the discriminant equation

[0109] The fingerprint of Psoralea corylifolia TCM granules and the fingerprint of Psoralea corylifolia salt TCM granules were compared to obtain common peaks. The ratio of peak area to sampling amount was used as the data for orthogonal partial least squares discriminant analysis. The marker peaks as differential characteristic indicators were determined according to the analysis results.

[0110] SPSS 20 software was used to perform discriminant analysis on the signature peaks of the differential characteristic indicators of multiple batches of samples used to build the model by selecting the default independent variable screening method "input independent variables together" in the discriminant analysis to establish a discriminant equation. In this application, the abbreviation of orthogonal partial least squares discriminant analysis is OPLS-DA, and the two have the same meaning.

[0111] Optionally, the fingerprint of the Psoralea corylifolia Chinese medicine formula granules and the fingerprint of the Psoralea corylifolia salt Chinese medicine formula granules can be obtained according to the above-mentioned fingerprint construction method.

[0112] In some embodiments, the software for performing orthogonal partial least squares discriminant analysis is SIMCA13.0.

[0113] In some embodiments, a total of 22 common peaks are obtained, and 9 marker peaks are determined based on the analysis results. SPSS20 software is used to perform discriminant analysis on the 9 marker peaks of differential characteristic indicators of multiple batches of samples used to build the model by selecting the default independent variable screening method "input independent variables together" in the discriminant analysis to establish a discriminant equation. The 9 marker peaks are characteristic peaks used as differential characteristic indicators. The marker peaks are selected based on the VIP value. VIP indicates the contribution of each substance to distinguishing samples. VIP>1 can be considered as the most significant indicator. Among the 22 common peaks, peak 3 psoralen, peak 5 isopsoralen, peak 12 psoralen, peak 14 isopsoralen, and peak 22 bakuchiol are identified chromatographic peaks. The retention times of the 22 common peaks are as follows: peak 1: 2.73±10%; peak 2: 3.56±10%; peak 3: 6.46±10%; peak 4: 7.01±10%; peak 5: 7.89±10%; peak 6: 9.01±10%; peak 7: 10. :9.55±10%; Peak 8:10.29±10%; Peak 9:11.88±10%; Peak 10:12.58±10%; Peak 11:14.79±10%; Peak 12:15.62±10%; Peak 13:16.33±10%; Peak 14:17.03±10%; Peak 15:17.54±10%; Peak 16:28.26±10%; Peak 17:28.99±10%; Peak 18:32.03±10%; Peak 19:32.48±10%; Peak 20:32.90±10%; Peak 21:36.57±10%; Peak 22:39.81±10%.

[0114] Furthermore, the differential characteristic indicators with VIP>1 are peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, and peak 4.

[0115] In some embodiments, the discriminant equation is Y=-0.008×peak 3-1.488×peak 4+0.056×peak 5+3.381×peak 9-3.863×peak 11-0.281×peak 12+1.220×peak 13+0.449×peak 14-1.367×peak 15-0.144. Wherein, peak 3 refers to the ratio of the peak area of ​​peak 3 to the sampling amount, and the rest are similar, for example, peak 5 refers to the ratio of the peak area of ​​peak 5 to the sampling amount.

[0116] S200: Obtaining the centroid value of Chinese medicine granules

[0117] The acquisition method includes: importing the data of peak area and sampling volume ratio of VIP>1 index (peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, peak 4) of multiple batches (for example, no less than 15 batches) of Psoralea corylifolia formula granules and salt Psoralea corylifolia formula granules samples into SPSS 20 software, selecting the default independent variable screening method "Enter independent variables together" in the discriminant analysis, and performing discriminant analysis.

[0118] In some embodiments, the centroid value of the Psoralea corylifolia Chinese medicine formula granules (centroid value I) is -3.992, and the centroid value of the Psoralea corylifolia salt Chinese medicine formula granules (centroid value II) is 3.992.

[0119] S300: Identification of the product to be tested

[0120] Extracting the Chinese medicine granules (test product) with an extraction solvent to prepare a test product solution;

[0121] The solution of the test product is subjected to ultra-high performance liquid chromatography, the ratio of the peak area of ​​the marker peak to the sampling amount is selected, substituted into the discriminant equation, and the discriminant score is calculated;

[0122] If the discrimination score is close to the centroid value I, it is judged as the Chinese medicine formula granules of Bu Gu Zhi, and if the discrimination score is close to the centroid value II, it is judged as the Chinese medicine formula granules of Salt Bu Gu Zhi;

[0123] The ultra-high performance liquid chromatography analysis includes elution according to the following mobile phase conditions: methanol is used as mobile phase A, and a formic acid aqueous solution with a volume concentration of 0.08% to 0.12% is used as mobile phase B, and the total volume percentage of mobile phase A and mobile phase B is 100%.

[0124] In some embodiments, in the identification method of Chinese medicine granules, the elution procedure includes:

[0125] From 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%;

[0126] From 9min to 18min, the volume percentage of the mobile phase A increases from 30% to 46%;

[0127] From 18min to 19min, the volume percentage of the mobile phase A increased from 46% to 57%;

[0128] From 19min to 34min, the volume percentage of the mobile phase A increased from 57% to 77%;

[0129] From 34min to 42min, the volume percentage of the mobile phase A increases from 77% to 90%;

[0130] 42min~42.1min, the volume percentage of the mobile phase A decreases from 90% to 18%;

[0131] From 42.1 min to 47 min, the volume percentage of the mobile phase A was maintained at 18%.

[0132] In some embodiments, in the identification method of Chinese medicine formula granules, the conditions of ultra-high performance liquid chromatography analysis include one or more of the following characteristics:

[0133] (1) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0134] (2) The model of the chromatographic column is ACQUITY UPLC BEH;

[0135] (3) Column temperature is 38°C to 42°C;

[0136] (4) The detection wavelength is 240nm to 250nm;

[0137] (5) The flow rate of the mobile phase is 0.28 mL / min to 0.32 mL / min.

[0138] In some embodiments, in the identification method of Chinese herbal formula granules, the extraction solvent is methanol and the extraction method is ultrasound.

[0139] The chromatographic conditions in the identification method of Chinese medicine formula granules are consistent with the chromatographic conditions constructed by fingerprint spectrum, which can reduce the detection error.

[0140] The following are some specific embodiments.

[0141] The experimental methods in the following examples are conventional methods unless otherwise specified. The medicinal materials, auxiliary materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified. Among them:

[0142] The instruments and models are as follows:

[0143] Thermo Vanquish ultra-high performance liquid chromatograph (Thermo Corporation), TUV detector (Thermo Corporation), Thermo workstation; ME204E one-ten-thousandth balance (Mettler Toledo Corporation); XP26 one-millionth balance (Mettler Toledo Corporation); 111B two-liang high-speed Chinese medicine pulverizer (Zhejiang Ruian Yongli Pharmaceutical Machinery Co., Ltd.); CNC KQ-500DE ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); MiliQ Direct 8 ultrapure water machine (Merck Millipore Corporation).

[0144] The raw materials and reagents are as follows:

[0145] Psoralea corylifolia Chinese medicine granules (batch numbers 101502, 15182, 1055312, 56522, 95852, 1030872, 1075132, 15181, 1075131, 1055321, 9066381, 1055311, 56601, 95851, 101501, 9066361, 9016391, 1030881, 95861) were provided by Guangdong Yifang Pharmaceutical Co., Ltd.

[0146] Salt Psoralea corylifolia Chinese medicine formula granules (batch numbers 57092, 8096082, 1041932, 1081062, 9016692, 96962, 116292, 96972, 9076382, 1031392, 1050092, 1031391, 96971, 1041931, 9076381, 1081051, 96961, 57021, 9016691, 8096071, 116291) were provided by Guangdong Yifang Pharmaceutical Co., Ltd.

[0147] Psoralen (mass fraction ≥ 98%, batch number ST55020105), isopsoralen (mass fraction ≥ 98%, batch number ST55030105), and bakuchiol (mass fraction ≥ 98.0%, batch number ST06860120) reference substances were purchased from Shanghai Shidande Standard Technical Service Co., Ltd.; psoralen (mass fraction ≥ 99.6%, batch number 110738-202016) and isopsoralen (mass fraction ≥ 99.4%, batch number 110739-201918) reference substances were purchased from China Food and Drug Inspection Institutes; methanol was chromatographic grade (Merck), formic acid was chromatographic grade (Tianjin Komiou Chemical Reagent Co., Ltd.), water was ultrapure water, and other reagents were of analytical grade.

[0148] Example 1 Construction of fingerprint

[0149] 1.1 Preparation of reference materials

[0150] Take appropriate amounts of psoralea corylifolia reference substance, isopsoralen reference substance, psoralen reference substance, isopsoralen reference substance, and bakuchiol reference substance, weigh them accurately, add methanol to dissolve them, and prepare a mixed solution containing 50 μg of psoralen, 50 μg of isopsoralen, 20 μg of psoralen, 20 μg of isopsoralen, and 100 μg of bakuchiol per 1 mL as a reference solution.

[0151] 1.2 Investigation of test sample preparation methods

[0152] 1.2.1 Investigation of extraction solvent

[0153] Take about 0.1g of sample powder, weigh it accurately, put it in a stoppered conical flask, accurately add 10% methanol, 30% methanol, 70% methanol, and 25mL of methanol, weigh it, and treat it with ultrasound (power 300W, frequency 45kHz) for 30min, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to get it. According to the chromatographic conditions determined under "1.3.5", inject the sample, measure it, and the results are shown in Figure 1 The results showed that the number of chromatographic peaks differed significantly when different extraction solvents were used, and the information of chromatographic peaks obtained by extraction with methanol and 70% methanol aqueous solution was equivalent. Considering the solvent extraction capacity and solution stability, methanol was a more suitable solvent, so methanol was determined as the extraction solvent.

[0154] 1.2.2 Investigation of extraction methods

[0155] Take about 0.1g of sample powder, weigh it accurately, put it in a stoppered conical flask, add 25mL of methanol accurately, weigh it, and treat it with ultrasound (power 300W, frequency 45kHz) for 30min, and heat it under reflux for 30min. Take it out, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate. According to the chromatographic conditions determined under "1.3.5", inject the sample, and measure it. The results are shown in Table 1. The results show that there is no obvious difference in "total peak area / sample weight" of different extraction methods. Considering the simplicity of operation, the extraction method is determined to be ultrasound.

[0156] Table 1 Results of investigation of different extraction methods

[0157]

[0158] 1.2.3 Extraction time investigation

[0159] Take about 0.1g of sample powder, weigh accurately, put it in a stoppered conical flask, add 25mL of methanol accurately, weigh it, and treat it with ultrasound (power 300W, frequency 45kHz) for 15, 30, and 45min, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to get it. According to the chromatographic conditions determined under "1.3.5", inject the sample, measure it, and the results are shown in Table 2. The results show that there is no obvious difference in "total peak area / sample weight" at different ultrasound times. In order to ensure complete extraction, the extraction time is determined to be 30min.

[0160] Table 2 Results of investigation at different extraction times

[0161]

[0162] 1.2.4 Investigation of extraction solvent dosage

[0163] Take about 0.1g of sample powder, weigh accurately, put it in a stoppered conical flask, accurately add 15, 25, and 50mL of methanol, weigh the weight, ultrasonically treat (power 300W, frequency 45kHz) for 30min, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to get it. According to the chromatographic conditions determined under "1.3.5", inject the sample, measure it, and the results are shown in Table 3. The results show that when the amount of extraction solvent is 15mL, 25mL, and 50mL, the numerical difference of the total characteristic peak area / sampling amount × solvent amount of the three characteristic spectra is not large. Based on the consideration of saving solvent, the amount of extraction solvent is determined to be 25mL.

[0164] Table 3 Results of investigation on different solvent dosages

[0165]

[0166] Based on the above investigation results, the preparation method of the test sample is determined as follows: take about 0.1g of the preparation granules, accurately weigh, place in a stoppered conical flask, add 25mL of methanol, weigh the weight, ultrasonically treat (power 300W, frequency 45kHz) for 30min, cool, filter, weigh again, make up the lost weight with methanol, shake well, filter, and take the filtrate.

[0167] 1.3 Establishing chromatographic conditions

[0168] 1.3.1 Determination of detection wavelength

[0169] Take about 0.1g of sample powder, weigh accurately, put it in a stoppered conical flask, add 25mL of methanol accurately, weigh the weight, ultrasonically treat (power 300W, frequency 45kHz) for 30min, take it out, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate. Elute according to the chromatographic gradient, column temperature, and flow rate determined under "1.3.5", record the chromatograms of the test solution at 246nm, 308nm, and 318nm respectively, compare the collected chromatograms, and determine the best wavelength. The results are as follows: Figure 2 The results show that the peak shapes of the chromatographic peaks at 246 nm are better than those at 308 nm and 318 nm, and the absorption intensity of the main chromatographic peaks is high, so the wavelength is selected as 246 nm.

[0170] 1.3.2 Column temperature investigation

[0171] Based on the experimental conditions proposed above, the column temperatures were investigated at 38°C, 40°C, and 42°C. The results are as follows: Figure 3 The results show that when the column temperature is 40℃, the chromatogram peak shape is better and the separation is moderate. Therefore, the column temperature is determined to be 40℃.

[0172] 1.3.3 Flow rate investigation

[0173] Based on the experimental conditions proposed above, the flow rates were investigated at 0.28 mL / min, 0.3 mL / min, and 0.32 mL / min. The results are as follows: Figure 4 The results show that when the flow rate is 0.3mL / min, the chromatogram peak shape is good and the separation is moderate. Therefore, the flow rate is determined to be 0.3mL / min.

[0174] 1.3.4 Choice of mobile phase

[0175] Take the same test solution and examine the mobile phases of acetonitrile-0.1% formic acid solution (B) and methanol-0.1% formic acid solution (B) respectively. According to the chromatographic conditions determined in "1.3.5", inject and measure. The results are as follows: Figure 5 The methanol-0.1% formic acid solution (B) system has relatively more peak information, better separation of each peak, and good peak shape of each component, so the methanol-0.1% formic acid solution (B) system was selected.

[0176] 1.3.5 Selection of elution gradient

[0177] Take the same test solution, use methanol (A)-0.1% formic acid solution (B) as the mobile phase, and elute according to the gradient in Table 4-6. Column temperature 40℃; detection wavelength 246nm; flow rate 0.3mL / min, injection volume 1μL. Results are shown in Figure 6 The results showed that the separation of the chromatographic peaks under gradient 1 was relatively better than that under gradient 2 and gradient 3, so gradient 1 with better separation was finally determined as the elution gradient of the characteristic spectrum.

[0178] Table 4 Gradient elution table

[0179]

[0180] Table 5 Gradient elution table

[0181]

[0182] Table 6 Gradient elution table

[0183]

[0184] Based on the above investigation results, the chromatographic conditions determined are as follows: ACQUITY UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm), column temperature 40 ° C, detection wavelength 246 nm. Methanol (A)-0.1% formic acid solution (B) was used as the mobile phase, eluted according to the elution program shown in Table 7, flow rate 0.3 mL / min, injection volume 1 μL.

[0185] Table 7 Elution program

[0186]

[0187] 1.4 Methodological considerations

[0188] 1.4.1 Precision test

[0189] Take the same test solution and inject it continuously for 6 times according to the chromatographic conditions determined under "1.3.5", record the peak area, take the psoralen chromatographic peak as the reference peak (S), and calculate the RSD values ​​of the relative retention time and relative peak area of ​​each characteristic peak, which are all less than 3%, indicating that the precision of the instrument is good.

[0190] 1.4.2 Repeatability test

[0191] Take 6 portions of the same batch of Psoralea corylifolia formula granule powder, weigh them accurately, prepare the test solution according to the test sample preparation method determined under "1.2", and measure them according to the chromatographic conditions determined under "1.3.5". Take the psoralen chromatographic peak as the reference peak (S), and calculate the RSD values ​​of the relative retention time and relative peak area of ​​each characteristic peak, which are all less than 3%, indicating that the method has good repeatability.

[0192] 1.4.3 Stability test

[0193] Accurately pipette the same test solution, and according to the chromatographic conditions determined under "1.3.5", inject and measure at 0, 2, 4, 6, 8, 10, 12, 18, and 24 h, respectively, and record the peak area. Taking the psoralen chromatographic peak as the reference peak (S), the RSD of the relative retention time and relative peak area of ​​each characteristic peak is calculated to be less than 3%, indicating that the test solution has good stability within 24 h.

[0194] 1.5 Establishment of characteristic spectra and determination of common peaks

[0195] 19 batches of Psoralea corylifolia formula granules and salt Psoralea corylifolia formula granules were taken respectively, and the test sample solution was prepared according to the test sample preparation method determined under "1.2", and the test was carried out according to the chromatographic conditions determined under "1.3.5", and the peak area of ​​each characteristic peak was recorded, and the peak area was normalized (i.e., the ratio of the peak area to the sampling amount was calculated). The results are shown in Tables 8-11. The characteristic spectrum data of Psoralea corylifolia formula granules and salt Psoralea corylifolia formula granules were respectively imported into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" software for processing, and the control characteristic spectrum of Psoralea corylifolia formula granules and the control characteristic spectrum of salt Psoralea corylifolia formula granules were established. The results are shown in Tables 8-11. Figure 7 , 8The characteristic spectrum marker established has 22 common peaks. Through the identification of reference substances, it was determined that peak 3, peak 5, peak 12, peak 14, and peak 22 are psoralen, isopsoralen, psoralen, isopsoralen, and bakuchiol, respectively. The results are shown in Fig. 9 .

[0196] Table 8 Normalization results of the peak areas of the common peaks in the characteristic spectra of 19 batches of Psoralea corylifolia granules (peaks 1 to 11)

[0197]

[0198] Table 9 Normalization results of the peak areas of the common peaks in the characteristic spectra of 19 batches of Psoralea corylifolia granules (peaks 12 to 22)

[0199]

[0200] Table 10 Normalization results of the peak areas of the common peaks in the characteristic spectra of 19 batches of salt psoralea formula granules (peaks 1 to 11)

[0201]

[0202] Table 11 Normalization results of the peak areas of the common peaks in the characteristic spectra of 19 batches of salt psoralea formula granules (peaks 12 to 22)

[0203]

[0204] 1.6 Orthogonal Partial Least Squares Discriminant Analysis

[0205] 1.6.1 Determine the differential characteristic index

[0206] After the peak areas of the chromatographic peaks in the fingerprints of 19 batches of Psoralea corylifolia and Psoralea corylifolia salt granules were normalized, orthogonal partial least squares discriminant analysis (OPLS-DA) was performed using SIMCA13.0 software to obtain the OPLS-DA score graph ( Fig.10 ). The Q2 of the model principal component regression coefficient is 0.9, which is greater than 0.5, indicating that the established prediction model is effective. The projection importance (VIP) results obtained by orthogonal partial least squares discriminant analysis are as follows Fig.11 According to the variable weight importance (VIP) in the model, factors with VIP values ​​> 1.0 can be considered as differential markers.

[0207] Depend on Fig.11 The factors with VIP values ​​> 1.0 include peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, and peak 4. These peaks can be regarded as differential characteristic indicators for distinguishing the granules of Psoralea corylifolia and Psoralea corylifolia salt formula.

[0208] 1.6.2 Discriminant Analysis

[0209] The VIP value greater than 1 index (peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, peak 4) obtained by OPLS-DA analysis was used as a variable, and the data of the ratio of the peak area and sampling volume of the VIP>1 index (peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, peak 4) of batches BGZ01~BGZ15 psoralea corylifolia and YBGZ01~YBGZ15 salt psoralea corylifolia formula granules were imported into SPSS 20 software, and the default independent variable screening method "input independent variables together" in the discriminant analysis was selected for discriminant analysis to establish a discriminant model. The results are shown in Tables 12-16. The results show that the p value of Wilks' Lambda is <0.05, indicating that the discriminant function is statistically significant. According to the typical discriminant function coefficients in Table 14, the unstandardized discriminant function formula can be obtained. The typical discriminant function formula of this example is as follows: Y = -0.008×Peak 3-1.488×Peak 4+0.056×Peak 5+3.381×Peak 9-3.863×Peak 11-0.281×Peak 12+1.220×Peak 13+0.449×Peak 14-1.367×Peak 15-0.144.

[0210] Table 12 Characteristic values

[0211] function Eigenvalue Variance (%) accumulation(%) Regular correlation 1 <![CDATA[17.081 a ]]> 100.0 100.0 0.972

[0212] In Table 12, a means that the first canonical discriminant function was used in the analysis.

[0213] In discriminant analysis, discriminant analysis is meaningful only when the group means are not equal. Wilks' lambda is the ratio of the sum of squares within the group to the total sum of squares. When the group means of all observations are equal, the Wilks' lambda value is 1; when the variation within the group is small compared to the total variation, the Wilks' lambda value is close to 0. Therefore, a large Wilks' lambda value indicates that the means of each group are basically equal; a small Wilks' lambda indicates that there are differences between groups.

[0214] Table 13 Ratio of the within-group sum of squares to the total sum of squares (Wilks' lambda)

[0215] Function Test Wilks' lambda Chi-square df Sig. 1 0.055 68.029 9 0.000

[0216] Table 14 Typical discriminant function system

[0217] function 1 Peak 3 -0.008 Peak 4 -1.488 Peak 5 0.056 Peak 9 3.381 Peak 11 -3.863 Peak 12 -0.281 Peak 13 1.220 Peak 14 0.449 Peak 15 -1.367 (constant) -0.144

[0218] Table 15 Functions at the centroid of the group

[0219] Group function 1 Psoralea corylifolia formula granules -3.993 Salt Psoralea corylifolia formula granules 3.993

[0220] Table 16 Classification results

[0221]

[0222] Example 2 Identification of samples with unknown components

[0223] 2.1 Sample preparation

[0224] Take an appropriate amount of the Chinese herbal formula granules to be tested, grind them into powder, take about 0.1g, weigh it accurately, put it in a stoppered conical flask, accurately add 25mL of methanol, weigh it, and treat it ultrasonically (power 300W, frequency 45kHz) for 30min. Let it cool, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain the test solution of the Chinese herbal formula granules to be tested.

[0225] 2.2 Testing process and results

[0226] Accurately pipette 1uL of the test solution of the Chinese medicine formula granules to be tested, inject it into the ultra-high performance liquid chromatograph, and measure according to the chromatographic conditions determined under "1.3.5" to obtain the peak areas of peak 9, peak 15, peak 3, peak 12, peak 5, peak 14, peak 11, peak 13, and peak 4, and calculate the ratio of peak area to sampling amount, and substitute it into the discriminant function Y = -0.008×peak 3-1.488×peak 4+0.056×peak 5+3.381×peak 9-3.863×peak 11-0.281×peak 12+1.220×peak 13+0.449×peak 14-1.367×peak 15-0.144, calculate the discrimination score, and then compare the distance with the centroid value of Psoralea corylifolia formula granules (-3.992) and the centroid value of Psoralea corylifolia salt formula granules (3.992), and the sample to be tested is judged to be Psoralea corylifolia formula granules if it is closer to the centroid value of Psoralea corylifolia formula granules (-3.992), otherwise it is judged to be Psoralea corylifolia salt formula granules. The discrimination results are shown in Table 17.

[0227] Table 17 Discrimination results

[0228]

[0229]

[0230] Among them, Group 1 represents Psoralea corylifolia formula granules; Group 2 represents Psoralea corylifolia salt formula granules.

[0231] As shown in Table 17, samples BGZ16 to BGZ19 were all judged to be psoralea corylifolia granules, and samples YBGZ16 to YBGZ21 were all judged to be salt psoralea corylifolia granules, which indicates that this discriminant function can effectively identify psoralea corylifolia granules and salt psoralea corylifolia granules with similar chemical components.

[0232] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples 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.

[0233] The above-described embodiments only express several implementation methods of the present application, but they cannot be understood as limiting the scope of the patent application. It should be pointed out that, for those of ordinary skill in the art, without departing from the concept of the present application, several variations and improvements can also be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art on the basis of the technical solutions provided in the present application through logical analysis, reasoning or limited experiments are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the attached claims, and the description and drawings may be used to explain the contents of the claims.

Claims

1. A method for constructing a fingerprint of Psoralea corylifolia or Psoralea corylifolia salt Chinese medicine granules, characterized in that: The steps include: Accurately weigh the Chinese medicine formula granules, extract them with an extraction solvent, and prepare a test solution; Accurately weigh the standard products of psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol, and dissolve them in a solvent to prepare a reference solution; Accurately aspirate the test sample solution and the reference solution, and perform ultra-high performance liquid chromatography analysis respectively to obtain a test sample fingerprint and a reference chromatogram, and obtain a fingerprint of the Chinese medicine formula granules according to the test sample fingerprint and the reference chromatogram; The conditions of the ultra-high performance liquid chromatography analysis include: (1) Using methanol as mobile phase A and using aqueous formic acid with a volume concentration of 0.08% to 0.12% as mobile phase B for gradient elution, the sum of the volume percentages of the mobile phase A and the mobile phase B is 100%, and the gradient elution procedure includes: from 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%, from 9 minutes to 18 minutes, the volume percentage of the mobile phase A increases from 30% to 46%, from 18 minutes to 19 minutes, the volume percentage of the mobile phase A increases from 46% to 57%, from 19 minutes to 34 minutes, the volume percentage of the mobile phase A increases from 57% to 77%, from 34 minutes to 42 minutes, the volume percentage of the mobile phase A increases from 77% to 90%, from 42 minutes to 42.1 minutes, the volume percentage of the mobile phase A decreases from 90% to 18%, and from 42.1 minutes to 47 minutes, the volume percentage of the mobile phase A is maintained at 18%; (2) The model of the chromatographic column is ACQUITY UPLC BEH; (3) Column temperature is 40°C; (4) Flow rate of 0.3 mL / min; and (5) The detection wavelength is 246nm.

2. The fingerprint construction method according to claim 1, characterized in that: The extraction solvent is methanol, and the extraction method is ultrasound.

3. The fingerprint construction method according to claim 1 or 2, characterized in that: The psoralea corylifolia Chinese medicine formula granules are prepared by comprising the following steps: Take the psoralea corylifolia slices, add water in an amount of 6 to 16 times the mass of the psoralea corylifolia slices, heat and boil for 0.5 to 2.5 hours; continue to add water in an amount of 5 to 12 times the mass of the psoralea corylifolia slices, heat and boil for 0.5 to 2.0 hours, filter, and concentrate the obtained filtrate under reduced pressure to a clear paste with a relative density of 1.02 to 1.10, filter, dry, and granulate.

4. The fingerprint construction method according to claim 1 or 2, characterized in that: The salt psoralea corylifolia Chinese medicine formula granules are prepared by comprising the following steps: Take salt psoralea corylifolia slices, add water in an amount of 6 to 16 times the mass of the salt psoralea corylifolia slices, heat and boil for 0.5 to 2.5 hours; continue to add water in an amount of 5 to 12 times the mass of the salt psoralea corylifolia slices, heat and boil for 0.5 to 2.0 hours, filter, and concentrate the obtained filtrate under reduced pressure to a clear paste with a relative density of 1.02 to 1.10, filter, dry, and granulate.

5. A method for identifying Chinese medicine granules, characterized in that: The steps include: According to the fingerprint construction method according to any one of claims 1 to 4, the fingerprint of the Psoralea corylifolia Chinese medicine formula granules and the fingerprint of the Psoralea corylifolia salt Chinese medicine formula granules are obtained; The fingerprint of the psoralea corylifolia Chinese medicine formula granules and the fingerprint of the salt psoralea corylifolia Chinese medicine formula granules were compared, and a total of 22 common peaks were calibrated. The ratio of the peak area and the sampling amount of each peak was used as the data for orthogonal partial least squares discriminant analysis. According to the analysis results, 9 marker peaks were determined, and discriminant analysis was performed using SPSS20 software based on the 9 marker peaks to establish a discriminant equation; wherein, the retention times of the 9 marker peaks are as follows: peak 9: 11.88±10%, peak 15: 17.54±10%, peak 3: 6.46±10%, peak 12: 15.62±10%, peak 5: 7.89±10%, peak 14: 17.03±10%, peak 11: 14.79±10%, peak 13: 16.33±10% and peak 4: 7.01±10%; Obtaining the centroid value I of the Psoralea corylifolia Chinese medicine formula granules and the centroid value II of the Psoralea corylifolia salt Chinese medicine formula granules; Extracting the Chinese medicine formula granules to be tested with an extraction solvent to prepare a test product solution; The test solution is subjected to ultra-high performance liquid chromatography to obtain the peak areas of the nine marker peaks, and the ratio of the peak areas of the nine marker peaks to the sampling amount is used as data, substituted into the discriminant equation, and the discriminant score is calculated; If the discrimination score is close to the centroid value I, it is judged to be the Psoralea corylifolia Chinese medicine formula granules; if the discrimination score is close to the centroid value II, it is judged to be the Salt Psoralea corylifolia Chinese medicine formula granules; Wherein, the conditions of the ultra-high performance liquid chromatography analysis include: (1) Methanol is used as mobile phase A, and aqueous formic acid solution with a volume concentration of 0.08% to 0.12% is used as mobile phase B, the sum of the volume percentages of the mobile phase A and the mobile phase B is 100%, and the gradient elution procedure includes: from 0 to 9 minutes, the volume percentage of the mobile phase A increases from 18% to 30%, from 9 minutes to 18 minutes, the volume percentage of the mobile phase A increases from 30% to 46%, from 18 minutes to 19 minutes, the volume percentage of the mobile phase A increases from 46% to 57%, from 19 minutes to 34 minutes, the volume percentage of the mobile phase A increases from 57% to 77%, from 34 minutes to 42 minutes, the volume percentage of the mobile phase A increases from 77% to 90%, from 42 minutes to 42.1 minutes, the volume percentage of the mobile phase A decreases from 90% to 18%, and from 42.1 minutes to 47 minutes, the volume percentage of the mobile phase A is maintained at 18%; (2) The model of the chromatographic column is ACQUITY UPLC BEH; (3) Column temperature is 40°C; (4) Flow rate of 0.3 mL / min; and (5) The detection wavelength is 246nm.

6. The identification method according to claim 5, characterized in that: The discriminant equation is: Y=-0.008×peak 3-1.488×peak 4+0.056×peak 5+3.381×peak 9-3.863×peak 11-0.281×peak 12+1.220×peak 13+0.449×peak 14-1.367×peak 15-0.144; In the discriminant equation, Y is the discriminant score, and peak 4, peak 5, peak 9, peak 11, peak 12, peak 13, peak 14 and peak 15 are the ratios of the peak area of ​​each peak to the sampling amount.

7. The identification method according to claim 5, characterized in that: The extraction solvent is methanol, and the extraction method is ultrasound.

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