A quality control method for a traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia and its application

Abstract

The present invention discloses a method for constructing a fingerprint spectrum of a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, and the method comprises the following steps: preparing a test solution; preparing a reference solution; and obtaining a fingerprint spectrum of the Chinese medicine compound according to the results of high performance liquid phase detection of the test solution and the reference solution. The present invention comprehensively and systematically analyzes the chemical components of the Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, and provides a theoretical basis for in-depth research on quality control and the material basis of drug efficacy.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a quality control method for a traditional Chinese medicine compound containing rehmannia root and psoralea corylifolia and an application thereof. Background Art

[0002] According to the Traditional Chinese Medicine Law of the People's Republic of China, ancient classic prescriptions refer to "prescriptions recorded in ancient Chinese medicine classics that are still widely used, have definite efficacy, and have obvious characteristics and advantages". Ancient classic prescriptions have a long and rich history of human use in my country and are still used today. They are the summary of the essence of clinical practice of doctors of all dynasties, carrying the profound accumulation of the splendid civilization of Chinese medicine for thousands of years. They are the result of thousands of years of tempering of Chinese medicine theory, the summary of clinical experience of all dynasties, and the most essential part of the great treasure house of Chinese medicine. In-depth research and development of classic Chinese medicine prescriptions is a golden key to tap into the treasure house of traditional Chinese medicine.

[0003] As the most commonly used dosage form of traditional Chinese medicine clinical medicine, decoctions of classic Chinese medicine prescriptions have the advantages of reasonable formulation, rapid onset, significant efficacy, and easy absorption, and are deeply trusted by the majority of patients. However, due to the shortcomings of easy mold and deterioration during preparation, carrying, temporary decoction, and long-term storage, the bitter taste and large amount of decoction, and the difficulty in standardization, which seriously affects its clinical efficacy, it cannot adapt to the living requirements of modern people. In order to maintain the advantages of decoctions and overcome the various shortcomings of decoctions, the "Technical Guiding Principles for Pharmaceutical Research of Traditional Chinese Medicine Compound Preparations Managed by the Catalog of Ancient Classic Prescriptions (Trial)" issued by the Drug Evaluation Center of the State Food and Drug Administration on August 31, 2021 clearly pointed out that the quality of traditional Chinese medicine compound preparations managed by the catalog of ancient classic prescriptions should be basically consistent with the quality of the classic prescription benchmark samples. The benchmark sample represents the overall intrinsic quality of the preparation. Except for the molding process, the benchmark sample and the other quality control indicators of the preparation are basically consistent. Therefore, the benchmark sample is a physical comparison of the intrinsic quality of the preparation and a reference for the optimization of large-scale production processes and the formulation of its quality standards. Benchmark samples are the benchmark for classic prescriptions and even all Chinese medicine research and development, and are the reference materials to ensure the safety and effectiveness of drugs. The production process route formulation, parameter optimization and quality standard formulation of classic prescription compound preparations should be based on the benchmark samples of classic prescriptions. Benchmark samples are the link between clinic, enterprise and scientific research, and are the benchmark for inheriting, applying and promoting traditional Chinese medicine.

[0004] Fingerprint is based on the understanding of the overall effect of Chinese medicine substance groups. It uses spectral and chromatographic techniques to obtain the spectrum or chromatogram of Chinese medicine chemical components. It is a feasible model for identifying the authenticity of Chinese medicine, evaluating quality consistency and product stability. It has the characteristics of large amount of information, strong characteristic, integrity and fuzziness. Chinese medicine fingerprint can comprehensively reflect the relative relationship of chemical components contained in medicinal materials, embody the complexity and correlation of Chinese medicine components, and adapt to the traditional theory of Chinese medicine. It can truly effectively characterize, comprehensively evaluate and comprehensively control the intrinsic quality of Chinese medicine. It is especially suitable for quality control of Chinese medicine and Chinese medicine products when the effective ingredients are not completely clear or do not need to be completely clear. In addition to being used to investigate the origin, harvesting season, harvesting part, processing, storage time and other factors of Chinese medicine, so as to provide a basis for identifying the authenticity and quality of raw materials before production, Chinese medicine fingerprint can also be used for quality control of Chinese medicine production process: tracking the changes of certain chemical components in preparations, monitoring the consistency and stability of quality between raw materials and finished products, and between batches of finished products. Compared with the quality analysis method of determining the content of index components, fingerprint spectrum can more comprehensively reflect the types and quantities of chemical components of traditional Chinese medicine. At the current situation where the effective ingredients of traditional Chinese medicine compound preparations have not been fully elucidated, it can realize the comprehensive evaluation of the intrinsic quality of traditional Chinese medicine and the effective control of its overall substance. It is one of the effective means of quality control of traditional Chinese medicine and its preparations.

[0005] At present, there is no literature report on a comprehensive analysis and fingerprint study of the chemical components of the traditional Chinese medicine compound comprising Rehmannia glutinosa and Psoralea corylifolia claimed in the present invention. In the prior art, only a single component in the traditional Chinese medicine compound is analyzed for quality, and there is no comprehensive and systematic quality control method to reflect the quality status of the main reference sample components in the traditional Chinese medicine compound and the finished product of the present invention, and it is impossible to effectively control its production process and product quality, and it is impossible to better guarantee its clinical efficacy. Therefore, it is necessary to adopt a fingerprint quality control method that can comprehensively control the overall quality of the traditional Chinese medicine compound of the present invention to control its key quality. Summary of the invention

[0006] Based on this, the present invention provides a fingerprint construction method for a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, and the construction method comprises the following steps: preparation of a test solution: taking an appropriate amount of a decoction of a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, shaking it well, filtering it, and taking a subsequent filtrate to obtain the test solution; wherein the Chinese medicine compound comprises Rehmannia glutinosa, Eucommia ulmoides, Psoralea corylifolia, Cuscuta australis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower and Angelica dahurica; preparation of a reference solution: weighing An appropriate amount of gallic acid, geniposide acid, and loganin reference substances are added with methanol to prepare a reference substance solution with a concentration of 10 to 100 μg / ml of gallic acid, geniposide acid, and loganin, respectively; a fingerprint spectrum of the Chinese medicine compound is obtained according to the results of high performance liquid phase detection of the test solution and the reference substance solution; the chromatographic conditions of the high performance liquid phase detection are: a chromatographic column with octadecylsilane bonded silica gel as filler is used, and a mobile phase A is selected from one or more of acetonitrile, methanol, and tetrahydrofuran , mobile phase B is acid aqueous solution, alkali aqueous solution and / or buffered saline solution, and the gradient elution program is: 0-15 min, 97% B→95% B; 15-20 min, 95% B→92% B; 20-25 min, 92% B→91% B; 25-30 min, 91% B→90% B; 30-35 min, 90% B; 35-40 min, 90% B→88% B; 40-55 min, 88% B→75% B ; 55-70 min, 75% B→63% B; 70-73 min, 63% B; 73-80 min, 63% B→62% B; 80-82 min, 62% B→50% B; 82-84 min, 50% B→97% B; 84-100 min, 97% B, the flow rate is 0.5-1.5 ml / min, the column temperature is 10-40°C, the detection wavelength is 190-400 nm, and the injection volume is 1-20 μl.

[0007] Further, the preparation method of the Chinese medicine compound decoction containing Rehmannia glutinosa and Psoralea corylifolia comprises: weighing appropriate amounts of Rehmannia glutinosa, Eucommia bark, Psoralea corylifolia, Cuscuta chinensis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower and Angelica dahurica in a container, adding water for soaking, boiling over high heat and then boiling over low heat, filtering through a filter to obtain the Chinese medicine compound decoction containing Rehmannia glutinosa and Psoralea corylifolia. Further, the container is a multifunctional decoction pot. Further, the sum of the mass of Rehmannia glutinosa, Eucommia bark, Psoralea corylifolia, Cuscuta chinensis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower and Angelica dahurica to the mass / volume (g / ml) of the water is 0.01 to 0.1, for example, about 0.061875. Further, the soaking time is 20 to 40 minutes, for example, about 30 minutes. Further, the slow fire decoction time is 40 to 60 minutes, for example, about 50 minutes. Further, the filter screen is a 150-mesh screen. Further, the volume of the Chinese medicine compound decoction containing Rehmannia glutinosa and Psoralea corylifolia is 400 to 700 ml, for example, about 550 ml. Further, the volume / volume ratio of the Chinese medicine compound decoction containing Rehmannia glutinosa and Psoralea corylifolia to the water is 0.4 to 0.8, for example, about 0.6875.

[0008] Furthermore, the mass ratios of the Rehmannia root, Eucommia bark, Psoralea corylifolia, Cuscuta australis, Myrrha sinensis, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Carthamus tinctorius and Angelica dahurica are (8-10):(2-4):(8-10):(8-10):(2-4):(2-4):(2-4):(2-4):(2-4):(0.5-2.5):(2-4). Furthermore, the mass ratios of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica are (8.5-9.5):(2.5-3.5):(8.5-9.5):(8.5-9.5):(2.5-3.5):(2.5-3.5):(2.5-3.5):(2.5-3.5):(2.5-3.5):(2.5-3.5):(1-2):(2.5-3.5). Further, the mass ratio between the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta seed, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica is about 9: about 3: about 9: about 9: about 3: about 3: about 3: about 3: about 3: about 3: about 1.5: about 3.

[0009] Further, the mass range of the Rehmannia glutinosa is 5 to 15g, for example, about 9g. Further, the mass range of the Eucommia ulmoides is 1 to 10g, for example, about 3g. Further, the mass range of the Psoralea corylifolia is 5 to 15g, for example, about 9g. Further, the mass range of the Cuscuta chinensis is 5 to 15g, for example, about 9g. Further, the mass range of the Myrrha is 1 to 10g, for example, about 3g. Further, the mass range of the Lycium barbarum is 1 to 10g, for example, about 3g. Further, the mass range of the Angelica sinensis is 1 to 10g, for example, about 3g. Further, the mass range of the Cornus officinalis is 1 to 10g, for example, about 3g. Further, the mass range of the Cistanche deserticola is 1 to 10g, for example, about 3g.

[0010] Further, the mass range of the safflower is 0.5-5g, for example, about 1.5g. Further, the mass range of the angelica dahurica is 1-10g, for example, about 3g.

[0011] Furthermore, in the method for preparing the test solution, the filtration is a microporous membrane filtration. Furthermore, the pore size of the microporous membrane is 0.22 μm.

[0012] Further, the concentration of gallic acid in the reference solution is about 50 μg / ml. Further, the concentration of geniposide in the reference solution is about 50 μg / ml. Further, the concentration of loganin in the reference solution is about 50 μg / ml.

[0013] Further, the flow rate of the high performance liquid phase detection is 0.8-1.2 ml / min, for example, about 1.0 ml / min. Further, the column temperature is 20-30°C, for example, about 25°C. Further, the detection wavelength is 230-370 nm, for example, about 254 nm. Further, the injection volume is 5-15 μl, for example, about 10 μl. Further, the theoretical plate number of the chromatographic peak corresponding to the loganin is not less than 10,000. Further, the chromatographic column is an Agilent 5TC-C18 chromatographic column. Further, the specifications of the chromatographic column are: column length 250 mm, inner diameter 4.6 mm, particle size 5 μm.

[0014] Further, mobile phase A is acetonitrile. Further, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution are selected from one or more of weak acids and their salts, weak bases and their salts of different concentrations. Further, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution are selected from formic acid, glacial acetic acid, phosphoric acid, trifluoroacetic acid, formic acid and ammonium formate, acetic acid and sodium acetate, acetic acid and ammonium acetate, disodium hydrogen phosphate and sodium dihydrogen phosphate, disodium hydrogen phosphate and potassium dihydrogen phosphate, disodium hydrogen phosphate and citric acid, citric acid and sodium citrate, glycine and hydrochloric acid, or phthalic acid and hydrochloric acid of different concentrations. Further, the acid aqueous solution is 0.01% to 1% acid aqueous solution. Further, the acid aqueous solution is 0.01% to 1% phosphoric acid aqueous solution. Further, the acid aqueous solution is about 0.1% phosphoric acid aqueous solution. Further, the buffered saline solution is acetate aqueous solution and / or acetate aqueous solution. Further, the pH value of the buffered saline solution is not more than 7.0.

[0015] Furthermore, when the detection wavelength is 254 nm, the fingerprint spectrum includes peaks 1-25, and the retention times of the peaks 1-25 correspond to about 4.777 min, about 5.337 min, about 9.123 min, about 11.799 min, about 12.369 min, about 14.580 min, about 16.771 min, about 19.373 min, about 21.382 min, about 21.893 min, about 22. 839 min, about 32.821 min, about 42.168 min, about 46.582 min, about 49.731 min, about 51.184 min, about 53.991 min, about 54.404 min, about 56.205 min, about 56.621 min, about 61.108 min, about 62.991 min, about 66.041 min, about 71.840 min and about 73.184 min.

[0016] Further, when the detection wavelength is 254nm, the fingerprint spectrum includes peaks 1-25, wherein peak 15 is used as a reference peak, and peaks 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24 and 25 are relative peaks. The average values ​​of retention time correspond to about 0.10, about 0.11, about 0.18, about 0.24, about 0.25, about 0.29, about 0.34, about 0.39, about 0.43, about 0.44, about 0.46, about 0.66, about 0.85, about 0.94, about 1.03, about 1.09, about 1.09, about 1.13, about 1.14, about 1.23, about 1.27, about 1.33, about 1.44 and about 1.47, respectively. Further, when the detection wavelength is 254 nm, peak 4 is gallic acid, peak 11 is geniposide acid, and peak 14 is loganin. Further, when the detection wavelength is 254nm, peak 1 comes from wolfberry fruit medicinal materials, peak 2 comes from safflower medicinal materials and Angelica dahurica medicinal materials, peak 3 comes from psoralea corylifolia medicinal materials, dodder seed medicinal materials, angelica medicinal materials and Angelica dahurica medicinal materials, peak 4 comes from Cornus officinalis medicinal materials, peak 5 comes from Rehmannia glutinosa medicinal materials, peak 6 comes from Rehmannia glutinosa medicinal materials, Eucommia ulmoides medicinal materials, Psoralea corylifolia medicinal materials and Cistanche deserticola medicinal materials, peak 7 comes from Rehmannia glutinosa medicinal materials and Cornus officinalis medicinal materials, peak 8 comes from Cuscuta chinensis medicinal materials, peak 9 comes from Rehmannia glutinosa medicinal materials, Eucommia ulmoides medicinal materials and Angelica dahurica medicinal materials, peak 10 comes from Myrrh medicinal materials and Cornus officinalis medicinal materials, peak No. 11 comes from Eucommia ulmoides medicinal materials, peak No. 12 comes from Cornus officinalis medicinal materials, peak No. 13 comes from Cuscuta australis medicinal materials, peak No. 14 comes from Cornus officinalis medicinal materials, peak No. 15 comes from Psoralea corylifolia medicinal materials, peak No. 16 comes from Psoralea corylifolia medicinal materials, peak No. 17 comes from Cuscuta australis medicinal materials, peak No. 18 comes from Cuscuta australis medicinal materials, peak No. 19 comes from Cuscuta australis medicinal materials, peak No. 20 comes from Cuscuta australis medicinal materials, peak No. 21 comes from Angelica sinensis medicinal materials, peak No. 22 comes from Angelica sinensis medicinal materials, peak No. 23 comes from Cuscuta australis medicinal materials, peak No. 24 comes from Psoralea corylifolia medicinal materials, and peak No. 25 comes from Psoralea corylifolia medicinal materials.

[0017] According to another aspect of the present invention, a quality control method for a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia is provided, the quality control method comprising the following steps: (1) establishing a standard fingerprint spectrum of a Chinese medicine compound reference sample according to the above-mentioned fingerprint spectrum construction method; (2) taking a test sample solution and testing it according to the chromatographic conditions in the above-mentioned fingerprint spectrum construction method to obtain a fingerprint spectrum of the Chinese medicine compound sample to be tested; and (3) comparing the fingerprint spectrum of the Chinese medicine compound sample to be tested obtained in step (2) with the standard fingerprint spectrum of the Chinese medicine compound reference sample obtained in step (1); a product that meets the requirements is a qualified product, and a product that does not meet the requirements is an unqualified product.

[0018] Furthermore, the compliance requirements include one or more of the following: (1) the fingerprint of the Chinese medicine compound sample to be tested presents 25 characteristic chromatographic peaks, and the retention time of each characteristic chromatographic peak is within ±10% of the retention time value of the corresponding reference chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; (2) taking peak No. 15 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine compound sample to be tested and the S peak is within ±10% of the relative retention time value of each characteristic chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; and (3) according to the Chinese medicine chromatographic fingerprint similarity evaluation system, the similarity between the fingerprint of the Chinese medicine compound sample to be tested and the standard fingerprint of the Chinese medicine compound reference sample shall not be less than 0.90 after similarity calculation.

[0019] According to another aspect of the present invention, there is provided a use of the above-mentioned construction method or the above-mentioned quality control method in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia.

[0020] Beneficial effects of the present invention:

[0021] In short, the present invention provides a fingerprint spectrum determination and quality control method for a Chinese medicine compound reference sample containing Rehmannia glutinosa and Psoralea corylifolia. The method confirms 25 common characteristic peaks, has simple conditions and short analysis time, solves the problems of difficulty in separating fingerprint characteristic peaks and interference from impurity peaks, ensures the chemical composition stability and safety of use of the reference sample, provides an important reference basis for the quality control of subsequent preparations, ensures the stable quality of the product, guarantees the efficacy of the Chinese medicine compound, and allows the Chinese medicine compound to better serve human life and health.

[0022] Specifically, compared with the prior art, the present invention has the following beneficial effects: (1) Establishing the fingerprint of the traditional Chinese medicine compound reference sample of the present invention overcomes the defect that the determination of the content of a single component is difficult to reflect the overall content, and can control the intrinsic quality of the traditional Chinese medicine compound reference sample of the present invention from a holistic and macroscopic perspective, thereby ensuring the efficacy of the drug, and using the main means of modern drug research to achieve the inheritance of fine products to classics, so that classic prescriptions are more formally controlled in terms of quality; (2) The chemical components in the traditional Chinese medicine compound reference sample of the present invention are complex, and it is difficult to separate its characteristic peaks. In the process of establishing the fingerprint, the present invention adopts a gradient elution method to solve the problem of difficulty in separating the fingerprint characteristic peaks and interference from impurity peaks; (3) In the process of establishing the fingerprint of the traditional Chinese medicine compound reference sample of the present invention, the present invention , 25 common characteristic peaks were confirmed, and their relative retention time, relative peak area and similarity were studied, which ensured the chemical composition stability and safety of the reference sample, and provided an important reference basis and quality reference for the quality control of the subsequent compound preparations; (4) The fingerprint spectrum of each effective ingredient in the traditional Chinese medicine compound reference sample of the present invention was regarded as a whole, and the sequence and relationship of each characteristic peak were emphasized, which not only avoided the one-sidedness of judging the overall quality of the traditional Chinese medicine compound reference sample of the present invention due to only measuring one or two chemical components, but also reduced the possibility of artificial processing for quality compliance, and provided a new method and means for completely and accurately evaluating the quality of the traditional Chinese medicine compound reference sample of the present invention; (5) The method of the present invention has good stability, high precision, good reproducibility, convenience and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0024] Figure 1 This is the reference fingerprint of the test solution of the Chinese medicine compound decoction of the present application at a wavelength of 254 nm, wherein peak 4: gallic acid; peak 11: geniposide acid; peak 14: loganin.

[0025] Figure 2 These are the fingerprint spectra of 15 batches of decoction of the Chinese herbal compound decoction of the present application at a wavelength of 254nm. DETAILED DESCRIPTION

[0026] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0027] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0028] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0029] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0030] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0031] As described in the background technology section, there is no literature report on the comprehensive analysis of the chemical components and fingerprint research of the Chinese medicine composition containing Rehmannia glutinosa and Psoralea corylifolia claimed by the present invention. In order to solve the above problems, the present invention provides a method for constructing a fingerprint of a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, and the construction method comprises the following steps: preparation of a test solution: take an appropriate amount of a decoction of a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia, shake well, filter, and take a filtrate to obtain the test solution; wherein the Chinese medicine compound includes Rehmannia glutinosa, Eucommia ulmoides, Psoralea corylifolia, Cuscuta australis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Carthamus tinctorius and Angelica dahurica; preparation of a reference solution : Weigh appropriate amounts of gallic acid, geniposide acid, and loganin reference substances, add methanol to prepare reference substance solutions with concentrations of gallic acid, geniposide acid, and loganin of 10 to 100 μg / ml, respectively; obtain a fingerprint of the Chinese medicine compound according to the results of high performance liquid chromatography detection of the test solution and the reference solution; the chromatographic conditions of the high performance liquid chromatography detection are: a chromatographic column with octadecylsilane bonded silica as filler, and a mobile phase A selected from one or more of acetonitrile, methanol, and tetrahydrofuran; Multiple, mobile phase B is acid aqueous solution, alkali aqueous solution and / or buffered saline solution, gradient elution program is: 0-15min, 97% B→95% B; 15-20min, 95% B→92% B; 20-25min, 92% B→91% B; 25-30min, 91% B→90% B; 30-35min, 90% B; 35-40min, 90% B→88% B; 40-55min, 88% B→75% B; 55-70 min, 75% B→63% B; 70-73 min, 63% B; 73-80 min, 63% B→62% B; 80-82 min, 62% B→50% B; 82-84 min, 50% B→97% B; 84-100 min, 97% B, the flow rate is 0.5-1.5 ml / min, the column temperature is 10-40°C, the detection wavelength is 190-400 nm, and the injection volume is 1-20 μl.

[0032] In the present invention, when concentration, time, flow rate, temperature, wavelength, volume, ratio, mass, or other values ​​or parameters are expressed as ranges, preferred ranges, or ranges defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether the range is disclosed separately. For example, when the range "10-40" is disclosed, the described range should be interpreted as including the range "10-40", "10-35", "10-30", "10-25", "10-20", "10-15", "15-40", "15-35", "15-30", "15-25", "15-20", "20-40", "20-35", "20-30", "20-25", "25-40", "25-35", "25-30", "30-40", "30-35", "35-40", etc. When a numerical range is described herein, unless otherwise indicated, that range is intended to include the endpoints and all integers and fractions within that range.

[0033] In a preferred embodiment, the preparation method of the Chinese herbal compound decoction containing Rehmannia root and Psoralea corylifolia comprises: weighing appropriate amounts of Rehmannia root, Eucommia bark, Psoralea corylifolia, Cuscuta australis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower and Angelica dahurica pubescens into a container, adding water for soaking, decocting with high heat until boiling, then decocting with low heat, filtering through a filter to obtain the Chinese herbal compound decoction containing Rehmannia root and Psoralea corylifolia.

[0034] In a preferred embodiment, the container is a multifunctional medicine decoction pot.

[0035] In a preferred embodiment, the ratio of the sum of the masses of Rehmannia glutinosa, Eucommia ulmoides, Psoralea corylifolia, Cuscuta australis, Myrrha myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Carthamus tinctorius and Angelica dahurica to the mass / volume (g / ml) of water is 0.01-0.1, for example, about 0.061875.

[0036] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.061875" includes ±5% of 0.061875, or from 0.05878125 to 0.06496875.

[0037] In a preferred embodiment, the soaking time is 20 to 40 minutes, such as about 30 minutes.

[0038] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 30" includes ±5% of 30, or from 28.5 to 31.5.

[0039] In a preferred embodiment, the slow-cooking time is 40 to 60 minutes, such as about 50 minutes.

[0040] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0041] In a preferred embodiment, the filter is a 150 mesh screen.

[0042] In a preferred embodiment, the volume of the Chinese herbal compound decoction containing Rehmannia root and Psoralea corylifolia is 400-700 ml, for example, about 550 ml.

[0043] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 550" includes ±5% of 550, or from 522.5 to 577.5.

[0044] In a preferred embodiment, the volume / volume ratio of the Chinese herbal compound decoction comprising Rehmannia root and Psoralea corylifolia to the water is 0.4-0.8, for example, about 0.6875.

[0045] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.6875" includes ±5% of 0.6875, or from 0.653125 to 0.721875.

[0046] In a preferred embodiment, the mass ratio of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica is (8-10):(2-4):(8-10):(8-10):(2-4):(2-4):(2-4):(2-4):(2-4):(0.5-2.5):(2-4). In a preferred embodiment, the mass ratio between the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica is (8.5-9.5): (2.5-3.5): (8.5-9.5): (8.5-9.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (1-2): (2.5-3.5). In a preferred embodiment, the mass ratio of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta seed, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica is about 9: about 3: about 9: about 9: about 3: about 3: about 3: about 3: about 3: about 3: about 1.5: about 3.

[0047] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 9" includes ±5% of 9, or from 8.55 to 9.45; "about 3" includes ±5% of 3, or from 2.85 to 3.15; "about 1.5" includes ±5% of 1.5, or from 1.425 to 1.575.

[0048] In a preferred embodiment, the mass of the Rehmannia glutinosa is in the range of 5 to 15 g, such as about 9 g.

[0049] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 9" includes ±5% of 9, or from 8.55 to 9.45.

[0050] In a preferred embodiment, the mass of the Eucommia ulmoides is in the range of 1 to 10 g, such as about 3 g.

[0051] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0052] In a preferred embodiment, the mass range of the Psoralea corylifolia is 5 to 15 g, for example, about 9 g.

[0053] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 9" includes ±5% of 9, or from 8.55 to 9.45.

[0054] In a preferred embodiment, the mass of the dodder seeds ranges from 5 to 15 g, such as about 9 g.

[0055] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 9" includes ±5% of 9, or from 8.55 to 9.45.

[0056] In a preferred embodiment, the mass of the myrrh is in the range of 1 to 10 g, such as about 3 g.

[0057] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0058] In a preferred embodiment, the mass of the wolfberry is in the range of 1 to 10 g, for example, about 3 g.

[0059] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0060] In a preferred embodiment, the mass of the angelica sinensis ranges from 1 to 10 g, such as about 3 g.

[0061] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0062] In a preferred embodiment, the weight of the Cornus officinalis is in the range of 1 to 10 g, such as about 3 g.

[0063] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0064] In a preferred embodiment, the mass of the Cistanche deserticola is in the range of 1 to 10 g, such as about 3 g.

[0065] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0066] In a preferred embodiment, the mass of safflower is in the range of 0.5 to 5 g, such as about 1.5 g.

[0067] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1.5" includes ±5% of 1.5, or from 1.425 to 1.575.

[0068] In a preferred embodiment, the mass of the Angelica dahurica is in the range of 1 to 10 g, for example, about 3 g.

[0069] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15.

[0070] In a preferred embodiment, in the method for preparing the test solution, the filtration is microporous membrane filtration.

[0071] In a preferred embodiment, the pore size of the microporous filter membrane is 0.22 μm.

[0072] In a preferred embodiment, the concentration of gallic acid in the reference solution is about 50 μg / ml.

[0073] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0074] In a preferred embodiment, the concentration of geniposide in the control solution is about 50 μg / ml.

[0075] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0076] In a preferred embodiment, the concentration of loganin in the reference solution is about 50 μg / ml.

[0077] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0078] In a preferred embodiment, the flow rate of the HPLC detection is 0.8-1.2 ml / min, for example, about 1.0 ml / min.

[0079] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1.0" includes ±5% of 1.0, or from 0.95 to 1.05.

[0080] In a preferred embodiment, the column temperature is 20-30°C, such as about 25°C.

[0081] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 25" includes ±5% of 25, or from 23.75 to 26.25.

[0082] In a preferred embodiment, the detection wavelength is 230-370 nm, for example, about 254 nm.

[0083] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 254" includes ±5% of 254, or from 241.3 to 266.7.

[0084] In a preferred embodiment, the injection volume is 5-15 μl, such as about 10 μl.

[0085] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 10" includes ±5% of 10, or from 9.5 to 10.5.

[0086] In a preferred embodiment, the theoretical plate number of the chromatographic peak corresponding to the loganin is not less than 10,000.

[0087] In a preferred embodiment, the chromatographic column is an Agilent 5TC-C18 chromatographic column.

[0088] In a preferred embodiment, the specifications of the chromatographic column are: column length 250 mm, inner diameter 4.6 mm, and particle size 5 μm.

[0089] In a preferred embodiment, mobile phase A is acetonitrile. In a preferred embodiment, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution are selected from one or more of weak acids and their salts, weak bases and their salts of different concentrations. In a preferred embodiment, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution are selected from formic acid, glacial acetic acid, phosphoric acid, trifluoroacetic acid, formic acid and ammonium formate, acetic acid and sodium acetate, acetic acid and ammonium acetate, disodium hydrogen phosphate and sodium dihydrogen phosphate, disodium hydrogen phosphate and potassium dihydrogen phosphate, disodium hydrogen phosphate and citric acid, citric acid and sodium citrate, glycine and hydrochloric acid, or phthalic acid and hydrochloric acid of different concentrations. In a preferred embodiment, the acid aqueous solution is 0.01% to 1% acid aqueous solution. In a preferred embodiment, the acid aqueous solution is 0.01% to 1% phosphoric acid aqueous solution. In a preferred embodiment, the acid aqueous solution is about 0.1% phosphoric acid aqueous solution.

[0090] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.1%" includes ±5% of 0.1%, or from 0.095% to 0.105%.

[0091] In a preferred embodiment, the buffered saline solution is an acetate solution and / or an acetate solution. In a preferred embodiment, the pH value of the buffered saline solution is not greater than 7.0.

[0092] In a preferred embodiment, when the detection wavelength is 254 nm, the fingerprint spectrum includes peaks 1-25, and the retention times of peaks 1-25 correspond to about 4.777 min, about 5.337 min, about 9.123 min, about 11.799 min, about 12.369 min, about 14.580 min, about 16.771 min, about 19.373 min, about 21.382 min, about 21.893 min, About 22.839 min, about 32.821 min, about 42.168 min, about 46.582 min, about 49.731 min, about 51.184 min, about 53.991 min, about 54.404 min, about 56.205 min, about 56.621 min, about 61.108 min, about 62.991 min, about 66.041 min, about 71.840 min and about 73.184 min.

[0093] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 4.777" includes ±5% of 4.777, or from 4.53815 to 5.01585; "about 5.337" includes ±5% of 5.337, or from 5.07015 to 5.60385; "about 9.123" includes ±5% of 9.123, or from 8.66685 to 9.57915; "about 11.799" includes ±5% of 11.799, or from 11.20905 to 12.38895; "about 12.369" includes ±5% of 12.369, or from 11.75055 to 12.98745; "about 14.580" includes ±5% of 14.580, or from 13.851 to 15.309; "about 16 ".771" includes ±5% of 16.771, or from 15.93245 to 17.60955; "about 19.373" includes ±5% of 19.373, or from 18.40435 to 20.34165; "about 21.382" includes ±5% of 21.382, or from 20.3129 to 22.4511; "about 21.893" includes ±5% of 21.893, or from 20.79835 to 22.98765; "about 22.839" includes ±5% of 22.839, or from 21.69705 to 23.98095; "about 32.821" includes ±5% of 32.821, or from 31.17995 to 34.4620 5; "about 42.168" includes ±5% of 42.168, or from 40.0596 to 44.2764; "about 46.582" includes ±5% of 46.582, or from 44.2529 to 48.9111; "about 49.731" includes ±5% of 49.731, or from 47.24445 to 52.21755; "about 51.184" includes ±5% of 51.184, or from 48.6248 to 53.7432; "about 53.991" includes ±5% of 53.991, or from 51.29145 to 56.69055; "about 54.404" includes ±5% of 54.404, or from 51.6838 to 57.124 2; "about 56.205" includes ±5% of 56.205, or from 53.39475 to 59.01525; "about 56.621" includes ±5% of 56.621, or from 53.78995 to 59.45205; "about 61.108" includes ±5% of 61.108, or from 58.0526 to 64.1634; "about 62.991" includes ±5% of 62.991, or from 59.84145 to 66.14055; "about 66.041" includes ±5% of 66.041, or from 62.73895 to 69.34305; "about 71.840" includes ±5% of 71.840, or from 68.248 to 75.432; "about 73.184" includes ±5% of 73.184, or from 69.5248 to 76.8432. .

[0094] In a preferred embodiment, when the detection wavelength is 254 nm, the fingerprint spectrum includes peaks 1-25, wherein peak 15 is used as a reference peak, peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, peak 14, peak 16, peak 17, peak 18, peak 19, peak 20, peak 21, peak 22, peak 23, peak 24 and peak 25 The relative retention time averages of the peaks are about 0.10, about 0.11, about 0.18, about 0.24, about 0.25, about 0.29, about 0.34, about 0.39, about 0.43, about 0.44, about 0.46, about 0.66, about 0.85, about 0.94, about 1.03, about 1.09, about 1.09, about 1.13, about 1.14, about 1.23, about 1.27, about 1.33, about 1.44 and about 1.47, respectively. In the present invention, "about" refers to a value within the range of ±5% of a specific value.For example, "about 0.10" includes ±5% of 0.10, or from 0.095 to 0.105; "about 0.11" includes ±5% of 0.11, or from 0.1045 to 0.1155; "about 0.18" includes ±5% of 0.18, or from 0.171 to 0.189; "about 0.24" includes ±5% of 0.24, or from 0.228 to 0.252; "about 0.25" includes ±5% of 0.25, or from 0.2375 to 0.2625; "about 0.29" includes ±5% of 0.29, or from 0.27 55 to 0.3045; "about 0.34" includes ±5% of 0.34, or from 0.323 to 0.357; "about 0.39" includes ±5% of 0.39, or from 0.3705 to 0.4095; "about 0.43" includes ±5% of 0.43, or from 0.4085 to 0.4515; "about 0.44" includes ±5% of 0.44, or from 0.418 to 0.462; "about 0.46" includes ±5% of 0.46, or from 0.437 to 0.483; "about 0.66" includes ±5% of 0.66 , or from 0.627 to 0.693; "about 0.85" includes ±5% of 0.85, or from 0.8075 to 0.8925; "about 0.94" includes ±5% of 0.94, or from 0.893 to 0.987; "about 1.03" includes ±5% of 1.03, or from 0.9785 to 1.0815; "about 1.09" includes ±5% of 1.09, or from 1.0355 to 1.1445; "about 1.13" includes ±5% of 1.13, or from 1.0735 to 1.1865; "about 1.14" includes ±5% of 1.14 “About 1.23” includes ±5% of 1.23, or from 1.1685 to 1.2915; “about 1.27” includes ±5% of 1.27, or from 1.2065 to 1.3335; “about 1.33” includes ±5% of 1.33, or from 1.2635 to 1.3965; “about 1.44” includes ±5% of 1.44, or from 1.368 to 1.512; “about 1.47” includes ±5% of 1.47, or from 1.3965 to 1.5435.

[0095] In a preferred embodiment, when the detection wavelength is 254 nm, peak 4 is gallic acid, peak 11 is geniposide acid, and peak 14 is loganin. In a preferred embodiment, when the detection wavelength is 254 nm, peak 1 is from Lycium barbarum medicinal material, peak 2 is from safflower medicinal material and Angelica dahurica medicinal material, peak 3 is from Psoralea corylifolia medicinal material, Cuscuta chinensis medicinal material, Angelica sinensis medicinal material and Angelica dahurica medicinal material, peak 4 is from Cornus officinalis medicinal material, peak 5 is from Rehmannia glutinosa medicinal material, peak 6 is from Rehmannia glutinosa medicinal material, Eucommia ulmoides medicinal material, Psoralea corylifolia medicinal material and Cistanche deserticola medicinal material, peak 7 is from Rehmannia glutinosa medicinal material and Cornus officinalis medicinal material, peak 8 is from Cuscuta chinensis medicinal material, peak 9 is from Rehmannia glutinosa medicinal material, Eucommia ulmoides medicinal material and Angelica dahurica medicinal material, peak 10 is from Myrrh Medicinal materials and Cornus officinalis medicinal materials, peak No. 11 comes from Eucommia ulmoides medicinal materials, peak No. 12 comes from Cornus officinalis medicinal materials, peak No. 13 comes from Cuscuta australis medicinal materials, peak No. 14 comes from Cornus officinalis medicinal materials, peak No. 15 comes from Psoralea corylifolia medicinal materials, peak No. 16 comes from Psoralea corylifolia medicinal materials, peak No. 17 comes from Cuscuta australis medicinal materials, peak No. 18 comes from Cuscuta australis medicinal materials, peak No. 19 comes from Cuscuta australis medicinal materials, peak No. 20 comes from Cuscuta australis medicinal materials, peak No. 21 comes from Angelica sinensis medicinal materials, peak No. 22 comes from Angelica sinensis medicinal materials, peak No. 23 comes from Cuscuta australis medicinal materials, peak No. 24 comes from Psoralea corylifolia medicinal materials, and peak No. 25 comes from Psoralea corylifolia medicinal materials.

[0096] According to another aspect of the present invention, a quality control method for a Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia is provided, the quality control method comprising the following steps: (1) establishing a standard fingerprint spectrum of a Chinese medicine compound reference sample according to the above-mentioned fingerprint spectrum construction method; (2) taking a test sample solution and testing it according to the chromatographic conditions in the above-mentioned fingerprint spectrum construction method to obtain a fingerprint spectrum of the Chinese medicine compound sample to be tested; and (3) comparing the fingerprint spectrum of the Chinese medicine compound sample to be tested obtained in step (2) with the standard fingerprint spectrum of the Chinese medicine compound reference sample obtained in step (1); a product that meets the requirements is a qualified product, and a product that does not meet the requirements is an unqualified product.

[0097] In a preferred embodiment, the compliance requirements include one or more of the following: (1) the fingerprint of the Chinese medicine compound sample to be tested presents 25 characteristic chromatographic peaks, and the retention time of each characteristic chromatographic peak is within ±10% of the retention time value of the corresponding reference chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; (2) taking peak No. 15 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine compound sample to be tested and the S peak is within ±10% of the relative retention time value of each characteristic chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; and (3) according to the Chinese medicine chromatographic fingerprint similarity evaluation system, the similarity between the fingerprint of the Chinese medicine compound sample to be tested and the standard fingerprint of the Chinese medicine compound reference sample shall not be less than 0.90 after similarity calculation.

[0098] According to another aspect of the present invention, there is provided a use of the above-mentioned construction method or the above-mentioned quality control method in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia.

[0099] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0100] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0101] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0102] Example

[0103] 1. Instruments and test drugs

[0104] Instruments: High performance liquid chromatograph Agilent 1290 chromatography system, including G1322A degasser, G1311A quaternary pump, G1313A autosampler, G1315B DAD diode array detector, G1316A column oven, ChemStation chromatography workstation; High performance liquid chromatograph Waters 2695 chromatography system, including quaternary gradient infusion pump (Alliance 2695), 120-position high performance autosampler, original imported chromatography column oven, Waters 2996 DAD diode array detector, Empower 3 chromatography management system; Chromatographic columns: Chromatographic column 1: Agilent 5TC-C185.0μm 4.6x250mm Serial No.731424; Chromatographic column 2: Agilent 5TC-C185.0μm 4.6x250mm Serial No.735516; Chromatographic column 3: Agilent 5TC-C185.0μm4.6x250mmSerialNo.735519; methanol, acetonitrile, phosphoric acid are chromatographically pure, glacial acetic acid and formic acid are analytically pure, and water is purified water. Reference substance information: loganin (batch number 111640-201808, content 99.0%, China Food and Drug Inspection Institute); geniposide (batch number 111828-201806, content 98.1%, China Food and Drug Inspection Institute); gallic acid (batch number 110831-201906, content 91.5%, China Food and Drug Inspection Institute).

[0105] 2. Solution preparation

[0106] 2.1. Preparation of reference solution

[0107] Take appropriate amount of gallic acid, geniposide acid and loganin reference substances respectively, weigh accurately, add methanol to make 1 ml of reference substance solution containing 50 μg of gallic acid, geniposide acid and loganin respectively, to make reference substance solution.

[0108] 2.2 Preparation of test solution

[0109] Rehmannia root 9g, Eucommia ulmoides 3g, Psoralea corylifolia 9g, Cuscuta australis 9g, Myrrha 3g, Lycium barbarum 3g, Angelica sinensis 3g, Cornus officinalis 3g, Cistanche deserticola 3g, Safflower 1.5g, and Angelica dahurica 3g, a total of 49.5g.

[0110] Preparation method: Take the above medicinal materials, put them in a multi-functional decoction pot, add 800ml of water to cover the medicinal materials by 2-5cm, soak for 30 minutes, boil over high heat, then simmer over low heat for about 50 minutes, filter with a 150-mesh sieve, and the final decoction volume should be about 550ml.

[0111] Take an appropriate amount of the above decoction, shake well, filter, and take the filtrate to obtain the product.

[0112] 2.3 Preparation of negative solution

[0113] 2.3.1. Preparation of negative sample solution lacking Rehmannia glutinosa

[0114] Weigh other medicinal ingredients according to the prescription, prepare a negative sample without Rehmannia glutinosa according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0115] 2.3.2 Preparation of negative sample solution lacking Eucommia ulmoides

[0116] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking Eucommia ulmoides according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0117] 2.3.3 Preparation of negative sample solution lacking Psoralea corylifolia

[0118] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking Psoralea corylifolia according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of Test Sample Solution".

[0119] 2.3.4. Preparation of negative sample solution lacking dodder seeds

[0120] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking Cuscuta australis according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0121] 2.3.5 Preparation of negative sample solution lacking myrrh

[0122] Weigh other medicinal ingredients according to the prescription, prepare a negative sample without myrrh according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0123] 2.3.6 Preparation of negative sample solution lacking wolfberry

[0124] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking wolfberry according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0125] 2.3.7. Preparation of negative sample solution lacking Angelica sinensis

[0126] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking Angelica sinensis according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0127] 2.3.8 Preparation of negative sample solution lacking Cornus officinalis

[0128] Weigh other medicinal ingredients according to the prescription, prepare a negative sample without Cornus officinalis according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0129] 2.3.9. Preparation of negative sample solution lacking Cistanche deserticola

[0130] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking Cistanche deserticola according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0131] 2.3.10. Preparation of negative sample solution lacking safflower

[0132] Weigh other medicinal ingredients according to the prescription, prepare a negative sample lacking safflower according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0133] 2.3.11. Preparation of Negative Sample Solution Lacking Angelica dahurica

[0134] Weigh other medicinal ingredients according to the prescription, prepare a negative sample of Duhuo according to the test sample preparation method, and prepare the negative sample solution according to "2.2, Preparation of test sample solution".

[0135] 2.4. Preparation of single-flavor solution

[0136] Take appropriate amounts of the eleven single-ingredient pieces of medicine respectively, put them into a stoppered conical flask, add 50 ml of water, heat and reflux for 50 minutes, cool, filter, and obtain the filtrate.

[0137] 3. Screening of chromatographic conditions

[0138] 3.1. Initial chromatographic conditions

[0139] Determined by high performance liquid chromatography (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia).

[0140] Chromatographic conditions and system suitability test: octadecyl bonded silica gel was used as the filler (Agilent 5TC-C18 5.0 μm 4.6x250 mm); acetonitrile was used as the mobile phase A, and 0.1% phosphoric acid was used as the mobile phase B; the flow rate was 1.0 ml / min, the column temperature was 25°C, and the initial gradient elution was performed according to the provisions in Table 1; the detection wavelength was 190-400 nm.

[0141] Table 1 Initial gradient

[0142] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0～120 5→95 95→5

[0143] According to the separation of components in the compound, the chromatographic column, the best absorption wavelength and the optimized gradient conditions are selected, and finally a better detection method is obtained.

[0144] The results show that from the isobsorption line diagram of Agilent 5TC-C18 at 190-400nm, there are fluorescence absorption points at wavelengths of 200-320nm. Comparison of the chromatograms at 210nm, 230nm, 254nm, 270nm, 290nm, 308nm, and 320nm shows that the response value of the chromatographic peak is optimal at 308nm, but its baseline is uneven. The response value and richness of the chromatographic peak at 254nm are better and the baseline is stable. Therefore, detection at wavelengths of 254nm and 308nm is determined, and the optimal wavelength is subsequently selected by comparison. There are chromatographic peaks in the first 60 minutes of the chromatogram, and there are basically no characteristic peaks afterwards. Therefore, considering that the acetonitrile ratio is within 50%, the gradient 1 in Table 2 is designed.

[0145] 3.2. Mobile phase gradient selection

[0146] In order to make each main chromatographic peak in the chromatogram have better separation parameters and present better chromatographic behavior, the test solution in "2.2, Preparation of test solution" was taken to optimize the elution gradient. The optimized gradient program is shown in Table 2.

[0147] Table 2 Gradient 1

[0148]

[0149]

[0150] The results showed that in the gradient 1 liquid chromatogram, the peak separation was poor in the 0-20 min period, and there was basically no characteristic peak after 45-65 min. Therefore, optimization was considered, and the column temperature was appropriately increased to 30°C to design the gradient 2 in Table 3.

[0151] Table 3 Gradient 2 (30°C)

[0152] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0～10 5→8 95→92 10～15 8→9 92→91 15～20 9→10 91→90 20～30 10→11 90→89 30～35 11→12 89→88 35～40 12→13 88→87 40～50 13→16 87→84 50～60 16→19 84→81 60～70 19→22 81→78 70～72 22→36 78→64 72～78 36→41 64→59 78～80 41→50 59→50 80～85 50→5 50→95 85～95 5 95

[0153] The results showed that there was basically no characteristic peak in the 60-70 min range of the gradient 2 chromatogram, and the characteristic peaks in the 70-85 min range had poor separation, so the gradient was further optimized and gradient 3 in Table 4 was designed.

[0154] Table 4 Gradient 3 (30°C)

[0155] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0～10 5→8 95→92 10～15 8→9 92→91 15～20 9→10 91→90 20～30 10→11 90→89 30～35 11→12 89→88 35～50 12→16 88→84 50～60 16→19 84→81 60～62 19→36 81→74 62～70 36→41 74→59 70～80 41→50 59→50 80～85 50→5 50→95 85～95 5 95

[0156] The results show that the characteristic peak separation of gradient 3 chromatogram is poor from 0 to 10 minutes, and the baseline is not stable and the peak separation is poor from 65 to 70 minutes, so it is considered to reduce the column temperature to 25 ° C, continue to optimize the gradient, and design gradient 4 in Table 5. Comparing the chromatogram at 254nm with the chromatogram at 308nm, it can be seen that the baseline of the chromatogram at 254nm is more stable than that at 308nm, and the peak richness and response value are better than those at a wavelength of 308nm, so the detection wavelength is determined to be 254nm.

[0157] Table 5 Gradient 4

[0158]

[0159]

[0160] The results showed that the separation of characteristic peaks at 0-10 minutes, 25-30 minutes, and 40-50 minutes in the gradient 4 chromatogram was poor, so the gradient was further optimized and gradient 5 in Table 6 was designed.

[0161] Table 6 Gradient 5

[0162] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0～15 5 95 15～20 5→8 95→92 20～25 8→9 92→91 25～30 9→10 91→90 30～35 10 90 35～50 10→25 90→75 50～60 25→37 75→63 60～70 37→38 63→62 70～75 38→40 62→60 75～80 40→50 60→50 80～85 50→5 50→95 85～95 5 95

[0163] The results showed that the separation of characteristic peaks at 0-15 minutes and 50-60 minutes in the gradient 5 chromatogram was poor, so the gradient was further optimized and gradient 6 in Table 7 was designed.

[0164] Table 7 Gradient 6

[0165]

[0166]

[0167] The results show that under the elution procedure of gradient 6, the baseline is stable, the main chromatographic peaks are well separated, and the analysis time is within 100 minutes. Considering that there are basically no characteristic peaks in the chromatogram of gradient 6 from 75 to 85 minutes and the equilibrium time period, the equilibrium time is extended and changed to gradient 7 in Table 8.

[0168] Table 8 Gradient 7

[0169] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0～15 3→5 97→95 15～20 5→8 95→92 20～25 8→9 92→91 25～30 9→10 91→90 30～35 10 90 35～40 10→12 90→88 40～55 12→25 88→75 55～70 25→37 75→63 70～73 37 63 73～80 37→38 63→62 80～82 38→50 62→50 82～84 50→3 50→97 84～100 3 97

[0170] The results showed that under gradient 7, the baseline was stable and the chromatographic peaks were well separated, which met the chromatographic requirements of the fingerprint. Therefore, this gradient was determined to be used for the fingerprint determination of the traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia.

[0171] Determined by high performance liquid chromatography (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia).

[0172] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel is used as filler; acetonitrile is used as mobile phase A, 0.1% phosphoric acid is used as mobile phase B, and gradient elution is performed according to the gradient 7 specified in Table 8; the flow rate is 1.0 ml / min, the column temperature is 25°C, and the detection wavelength is 254 nm. The theoretical plate number calculated based on the loganin peak should not be less than 10,000.

[0173] Preparation of test solution: Take an appropriate amount of the Chinese medicine compound sample (decoction) containing Rehmannia root and Psoralea corylifolia, and filter through 0.22μm to obtain the solution.

[0174] Determination method: Accurately pipette 10μl of the test solution, inject it into the liquid chromatograph, and determine it.

[0175] 4. Validation of analytical methods

[0176] The fingerprint spectrum established above was systematically methodologically validated in accordance with the 9101 Analytical Method Validation Guidelines of Part IV of the 2020 Edition of the Chinese Pharmacopoeia and the National Drug Standard Work Manual (Fourth Edition).

[0177] 4.1. Exclusivity

[0178] Prepare negative blank sample solution according to the test solution preparation method, test it according to the fingerprint method, and record the chromatogram.

[0179] By comparing the absorption spectrum with that of the reference solution, it was determined that peak 4, peak 11 and peak 14 were gallic acid, geniposide acid and loganin, respectively, as shown in Table 9.

[0180] Table 9 System parameters of the fingerprint of the traditional Chinese medicine compound containing Radix Rehmanniae Preparata and Psoralea corylifolia

[0181]

[0182]

[0183] The results showed that the chromatogram of the Chinese medicine compound benchmark sample containing Rehmannia glutinosa and Psoralea corylifolia mainly showed 25 peaks, peak 1 belonged to Lycium barbarum; peak 2 belonged to Carthamus tinctorius and Angelica dahurica; peak 3 belonged to Psoralea corylifolia, Cuscuta chinensis, Angelica sinensis and Angelica dahurica; peaks 4, 12 and 14 belonged to Cornus officinalis, peak 4 was gallic acid, peak 14 was loganin; peak 5 belonged to Rehmannia glutinosa; peak 6 belonged to Rehmannia glutinosa, Eucommia ulmoides, Psoralea corylifolia and Cistanche deserticola; Peak 7 belongs to Radix Rehmanniae Preparata and Fructus Corni; Peak 8, Peak 13, Peak 17-Peak 20, Peak 23 belong to Semen Cuscutae; Peak 9 belongs to Radix Rehmanniae Preparata, Eucommia ulmoides, and Angelica dahurica; Peak 10 belongs to Myrrha and Fructus Corni; Peak 11 belongs to Eucommia ulmoides, and Peak 11 is Geniposide; Peak 15-Peak 16, Peak 24-Peak 25 belong to Fructus Psoraleae; Peak 21-Peak 22 belong to Angelica sinensis, negative without interference, and good method specificity. The specific summary is shown in Table 10.

[0184] Table 10 Summary of specific chromatographic peaks of the Chinese medicinal compound containing Radix Rehmanniae Preparata and Fructus Psoraleae of the present invention

[0185]

[0186]

[0187] 4.2 Precision

[0188] 4.2.1. Instrument precision test

[0189] The same test solution was sampled 6 times continuously, the chromatogram was recorded, and the similarity of each characteristic peak was calculated. The results showed that the similarity of the instrument precision of this method was greater than 0.90, indicating that the instrument precision of this method was good.

[0190] 4.2.2 Intermediate precision

[0191] Two personnel prepared 6 test solutions according to the above conditions, respectively, injected and tested on Agilent HPLC and Waters HPLC, recorded chromatograms, and calculated the similarity of each characteristic peak. The results showed that the similarity of the intermediate precision of each characteristic peak was greater than 0.90, which met the requirements.

[0192] 4.2.3 Repeatability test

[0193] Take 6 samples from the same batch, prepare the test solution according to the method of "2.2, Preparation of Test Solution", record the chromatogram, and calculate the similarity of each characteristic peak. The results show that the similarity of the repeatability of each characteristic peak is greater than 0.90, which meets the requirements.

[0194] 4.3 Durability

[0195] 4.3.1 Stability Study

[0196] Take the same test solution and inject it at 0, 3h, 6h, 9h, 17h, 24h, 36h and 48h. Record the chromatogram, calculate the similarity of each characteristic peak, and examine the stability of the test solution. The results show that within 48h of stability, the similarity of each characteristic peak is greater than 0.90, which meets the requirements, indicating that it is stable within 48h.

[0197] 4.3.2 Investigation of different flow rates

[0198] Take the same test solution and test it at different flow rates of 1.0±0.1ml / min, record the chromatogram, and calculate the similarity of each characteristic peak. The results show that the similarity results of different flow rates are all greater than 0.90, and the flow rate durability is good and meets the regulations.

[0199] 4.3.3 Investigation of different column temperatures

[0200] Take the same test solution, test at 35±3℃, record the chromatogram, calculate the similarity of each characteristic peak, and perform similarity matching calculation with the standard reference spectrum. The results show that the similarity of each characteristic peak is greater than 0.90, and the column temperature durability is good and meets the requirements.

[0201] 4.3.4. Investigation of different chromatographic columns

[0202] The same test solution was tested on different chromatographic columns (chromatographic column 1: Agilent 5TC-C18 5.0μm 4.6x250mm No.731424), (chromatographic column 2: Agilent 5TC-C18 5.0μm 4.6x250mm No.735516), (chromatographic column 3: Agilent 5TC-C18 5.0μm 4.6x250mm No.735519), and the chromatograms were recorded and the similarity matching calculation was performed with the standard reference spectrum of the traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia. The results showed that the similarity results of different chromatographic columns were all greater than 0.90, which met the requirements.

[0203] 4.3.5. Investigation of mobile phases with different acid types

[0204] The results showed that under the condition of 0.1% formic acid, the spectrum t R =42.168 and t R =53.991 characteristic peak is lost, and the spectrum is t R =21.893 and t R The characteristic peak of =22.839 was lost, so phosphoric acid was used as the acid type of the mobile phase.

[0205] 4.3.6. Investigation of mobile phases with different acid concentrations

[0206] The same test solution was tested at 0.09%, 0.10% and 0.11% phosphoric acid solution concentration in the mobile phase, and the chromatogram was recorded and the similarity matching calculation was performed with the standard reference spectrum of the traditional Chinese medicine compound containing Rehmannia glutinosa and Psoralea corylifolia. The results showed that under different acid concentration conditions, the similarity results of each common peak were greater than 0.90, which met the requirements.

[0207] 4.4. Fingerprint method confirmation

[0208] According to the results of the above methodological investigation, among the 25 common peaks of the established fingerprint, each chromatographic peak is less affected by factors such as different brands of HPLC, different chromatographic columns, different column temperatures and flow rates, and the similarities of each characteristic peak of the instrument precision, intermediate precision, repeatability test and stability test are all greater than 0.9, indicating that the method has good durability, is stable and reliable, and can be used for fingerprint determination.

[0209] Based on the above results, the fingerprint method is as follows:

[0210] Determined by high performance liquid chromatography (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia).

[0211] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel is used as filler; acetonitrile is used as mobile phase A, 0.1% phosphoric acid is used as mobile phase B; gradient elution is performed according to the provisions of gradient 7 in Table 8; the flow rate is 1.0 ml / min, the column temperature is 25°C, and the detection wavelength is 254 nm. The theoretical plate number calculated based on the loganin peak should not be less than 10,000.

[0212] Preparation of reference solution: Take appropriate amount of gallic acid, geniposide acid and loganin reference substances respectively, weigh accurately, add methanol to make 1 ml of reference substance solution containing 50 μg of gallic acid, geniposide acid and loganin respectively.

[0213] Preparation of test solution: Take an appropriate amount of the Chinese medicine compound sample (decoction) containing Rehmannia glutinosa and Psoralea corylifolia, and filter it through a 0.22μm microporous filter membrane to obtain the solution.

[0214] Determination method: Accurately pipette 10μl of the test solution, inject it into the liquid chromatograph, and determine it.

[0215] 5. Determination of fingerprint standard

[0216] 15 batches of multi-batch matching pictures Figure 2 It is determined that there are 25 characteristic peaks in the chromatogram of the test sample, among which the peak corresponding to peak 15 is the S peak. According to the similarity evaluation system of traditional Chinese medicine chromatographic fingerprint, the similarity between the fingerprint of the test sample and the fingerprint of the control shall not be less than 0.90.

[0217] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A method for constructing a fingerprint of a Chinese medicinal compound containing Rehmannia glutinosa and Psoralea corylifolia, characterized in that: The construction method comprises the following steps: Preparation of the test solution: taking an appropriate amount of a decoction of a Chinese medicinal compound containing Rehmannia glutinosa and Psoralea corylifolia, shaking it well, filtering it, and taking a subsequent filtrate to obtain the test solution; wherein the Chinese medicinal compound is composed of Rehmannia glutinosa, Eucommia ulmoides, Psoralea corylifolia, Cuscuta australis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower, and Angelica dahurica; Preparation of reference solution: Weigh appropriate amounts of gallic acid, geniposide, and loganin reference substances, add methanol to prepare reference solutions with concentrations of gallic acid, geniposide, and loganin of 10 to 100 μg / ml, respectively; Obtaining a fingerprint of the Chinese medicine compound according to the results of high performance liquid chromatography detection of the test solution and the reference solution; The chromatographic conditions of the high performance liquid phase detection are as follows: a chromatographic column with octadecylsilane bonded silica as filler is used, the chromatographic column is an Agilent 5TC-C18 chromatographic column, the specifications of the chromatographic column are as follows: column length 250 mm, inner diameter 4.6 mm, particle size 5 μm, mobile phase A is acetonitrile, mobile phase B is 0.09-0.11% phosphoric acid aqueous solution, and the gradient elution program is as follows: 0-15 min, 97% B→95% B; 15-20 min, 95% B→92% B; 20-25 min, 92% B→91% B; 25-30 min, 91% B→90% B; 30-35 min, 90% B; 35-40 min, 9 0% B→88% B; 40~55 min, 88% B→75% B; 55~70 min, 75% B→63% B; 70~73 min, 63% B; 73~80 min, 63% B→62% B; 80~82 min, 62% B→50% B; 82~84 min, 50% B→97% B; 84~100 min, 97% B, the flow rate is 0.9~1.1 ml / min, the column temperature is 25°C, the detection wavelength is 254 nm, and the injection volume is 1~20 μl.

2. The construction method according to claim 1, characterized in that: The preparation method of the Chinese medicinal compound decoction containing Rehmannia root and Psoralea corylifolia comprises: weighing appropriate amounts of Rehmannia root, Eucommia bark, Psoralea corylifolia, Cuscuta chinensis, Myrrh, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Safflower and Angelica dahurica in a container, adding water for soaking, decocting with high heat until boiling, then decocting with low heat, filtering through a filter to obtain the Chinese medicinal compound decoction containing Rehmannia root and Psoralea corylifolia.

3. The construction method according to claim 2, characterized in that: The container is a multifunctional medicine decocting pot.

4. The construction method according to claim 2, characterized in that: The ratio of the sum of the masses of the Radix Rehmanniae, Eucommia ulmoides, Psoralea corylifolia, Cuscuta australis, Myrrha sinensis, Fructus Lycii, Angelica sinensis, Fructus Corni, Cistanche deserticola, Carthamus tinctorius and Angelica dahurica to the mass / volume (g / ml) of the water is 0.01-0.

1.

5. The construction method according to claim 4, characterized in that: The ratio of the sum of the masses of the Rehmannia root, Eucommia bark, Psoralea corylifolia, Cuscuta australis, Myrrha sinensis, Lycium barbarum, Angelica sinensis, Cornus officinalis, Cistanche deserticola, Carthamus tinctorius and Angelica dahurica to the mass / volume (g / ml) of the water is 0.05878125-0.06496875.

6. The construction method according to claim 2, characterized in that: The soaking time is 20 to 40 minutes.

7. The construction method according to claim 6, characterized in that: The soaking time is 28.5 to 31.5 minutes.

8. The construction method according to claim 2, characterized in that: The slow fire decocting time is 40 to 60 minutes.

9. The construction method according to claim 8, characterized in that: The slow fire decocting time is 47.5 to 52.5 minutes.

10. The construction method according to claim 2, characterized in that: The filter is a 150-mesh screen.

11. The construction method according to claim 2, characterized in that: The volume of the Chinese medicinal compound decoction containing Rehmannia root and Psoralea corylifolia is 400-700 ml.

12. The construction method according to claim 11, characterized in that: The volume of the Chinese medicinal compound decoction containing Rehmannia root and Psoralea corylifolia is 522.5-577.5 ml.

13. The construction method according to claim 2, characterized in that: The volume / volume ratio of the traditional Chinese medicine compound decoction containing Rehmannia root and Psoralea corylifolia to the water is 0.4-0.

8.

14. The construction method according to claim 13, characterized in that: The volume / volume ratio of the traditional Chinese medicine compound decoction containing Rehmannia root and Psoralea corylifolia to the water is 0.653125-0.721875.

15. The construction method according to claim 1, characterized in that: The mass ratio of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha sinensis, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica is (8-10): (2-4): (8-10): (8-10): (2-4): (2-4): (2-4): (2-4): (2-4): (0.5-2.5): (2-4).

16. The construction method according to claim 15, characterized in that: The mass ratios of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha myrrh, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica are (8.5-9.5): (2.5-3.5): (8.5-9.5): (8.5-9.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (2.5-3.5): (1-2): (2.5-3.5).

17. The construction method according to claim 16, characterized in that: The mass ratios of the Rehmannia root, the Eucommia bark, the Psoralea corylifolia, the Cuscuta chinensis, the Myrrha myrrh, the Lycium barbarum, the Angelica sinensis, the Cornus officinalis, the Cistanche deserticola, the Carthamus tinctorius and the Angelica dahurica are (8.55-9.45): (2.85-3.15): (8.55-9.45): (8.55-9.45): (2.85-3.15): (2.85-3.15): (2.85-3.15): (2.85-3.15): (2.85-3.15): (2.85-3.15): (1.425-1.575): (2.85-3.15).

18. The construction method according to claim 1, characterized in that: The mass range of the Rehmannia glutinosa is 5 to 15 g.

19. The construction method according to claim 18, characterized in that: The mass range of the Rehmannia glutinosa is 8.55-9.45g.

20. The construction method according to claim 1, characterized in that: The mass range of the Eucommia ulmoides is 1 to 10 g.

21. The construction method according to claim 20, characterized in that: The mass range of the Eucommia ulmoides is 2.85-3.15g.

22. The construction method according to claim 1, characterized in that: The mass range of the psoralea corylifolia is 5 to 15 g.

23. The construction method according to claim 22, characterized in that: The mass range of the psoralea corylifolia is 8.55-9.45g.

24. The construction method according to claim 1, characterized in that: The mass range of the dodder seeds is 5 to 15 g.

25. The construction method according to claim 24, characterized in that: The mass range of the dodder seeds is 8.55 to 9.45 g.

26. The construction method according to claim 1, characterized in that: The mass range of the myrrh is 1 to 10 g.

27. The construction method according to claim 26, characterized in that: The mass range of the myrrh is 2.85 to 3.15 g.

28. The construction method according to claim 1, characterized in that: The mass range of the wolfberry is 1 to 10 g.

29. The construction method according to claim 28, characterized in that: The mass range of the wolfberry is 2.85 to 3.15 g.

30. The construction method according to claim 1, characterized in that: The mass range of the angelica is 1 to 10 g.

31. The construction method according to claim 30, characterized in that: The mass range of the angelica is 2.85 to 3.15 g.

32. The construction method according to claim 1, characterized in that: The weight range of the Cornus officinalis is 1 to 10 g.

33. The construction method according to claim 32, characterized in that: The mass range of the Cornus officinalis is 2.85-3.15g.

34. The construction method according to claim 1, characterized in that: The mass range of the Cistanche deserticola is 1 to 10 g.

35. The construction method according to claim 34, characterized in that: The mass range of the Cistanche deserticola is 2.85-3.15g.

36. The construction method according to claim 1, characterized in that: The mass range of the safflower is 0.5 to 5 g.

37. The construction method according to claim 36, characterized in that: The mass range of the safflower is 1.425-1.575g.

38. The construction method according to claim 1, characterized in that: The mass range of the Angelica dahurica is 1 to 10 g.

39. The construction method according to claim 38, characterized in that: The mass range of the Angelica dahurica is 2.85 to 3.15 g.

40. The construction method according to claim 1, characterized in that: In the method for preparing the test solution, the filtration is microporous membrane filtration.

41. The construction method according to claim 40, characterized in that: The pore size of the microporous filter membrane is 0.22 μm.

42. The construction method according to claim 1, characterized in that: The concentration of gallic acid in the reference solution is 47.5-52.5 μg / ml.

43. The construction method according to claim 1, characterized in that: The concentration of geniposide in the reference solution is 47.5-52.5 μg / ml.

44. The construction method according to claim 1, characterized in that: The concentration of loganin in the reference solution is 47.5-52.5 μg / ml.

45. The construction method according to claim 1, characterized in that: The injection volume is 5 to 15 μl.

46. ​​The construction method according to claim 45, characterized in that The injection volume is 9.5-10.5 μl.

47. The construction method according to claim 1, characterized in that The theoretical plate number of the chromatographic peak corresponding to the loganin is not less than 10,000.

48. The construction method according to claim 1, characterized in that: When the detection wavelength is 254 nm, the fingerprint spectrum includes peaks 1 to 25, and the retention times of peaks 1 to 25 correspond to 4.53815-5.01585 min, 5.07015-5.60385 min, 8.66685-9.57915 min, 11.20905-12.38895 min, 11.75055-12.98745 min, 13.851-15.309 min, 15.93245-17.60955 min, 18.40435-20.34165 min, 20.3129-22.4511 min, 20.79835-22.98765 min, 21.69705-23.98745 min, 31.17 995~34.46205min, 40.0596~44.2764min, 44.2529~48.9111min, 47.24445~52.21 755min, 48.6248~53.7432min, 51.29145~56.69055min, 51.6838~57.1242min, 53 .39475~59.01525min, 53.78995~59.45205min, 58.0526~64.1634min, 59.84145~66.14055min, 62.73895~69.34305min, 68.248~75.432min and 69.5248~76.8432min.

49. The construction method according to claim 1, characterized in that: When the detection wavelength is 254 nm, the fingerprint spectrum includes peaks 1-25, wherein peak 15 is used as a reference peak, and the relative retention time averages of peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, peak 14, peak 16, peak 17, peak 18, peak 19, peak 20, peak 21, peak 22, peak 23, peak 24 and peak 25 are 0.095-0.105, 0.1045-0.1155, 0.171-0.189, 0.228-0.252, 0.2375-0.2625, 0.2755-0.3 0.45, 0.323-0.357, 0.3705-0.4095, 0.4085-0.4515, 0.418-0.462, 0.437-0.483, 0.627-0.693, 0.8075-0.8925, 0.893-0.987, 0.9785-1.0815, 1.0355-1.1445, 1.0355-1.1445, 1.0735-1.1865, 1.083-1.197, 1.1685-1.2915, 1.2065-1.3335, 1.2635-1.3965, 1.368-1.512 and 1.3965-1.5435.

50. The construction method according to claim 1, characterized in that: When the detection wavelength is 254 nm, peak No. 4 is gallic acid, peak No. 11 is geniposide acid, and peak No. 14 is loganin.

51. The construction method according to claim 1, characterized in that: When the detection wavelength is 254nm, peak 1 comes from wolfberry fruit medicinal materials, peak 2 comes from safflower medicinal materials and Angelica dahurica medicinal materials, peak 3 comes from psoralea corylifolia medicinal materials, dodder seed medicinal materials, angelica medicinal materials and Angelica dahurica medicinal materials, peak 4 comes from Cornus officinalis medicinal materials, peak 5 comes from Rehmannia glutinosa medicinal materials, peak 6 comes from Rehmannia glutinosa medicinal materials, Eucommia ulmoides medicinal materials, Psoralea corylifolia medicinal materials and Cistanche deserticola medicinal materials, peak 7 comes from Rehmannia glutinosa medicinal materials and Cornus officinalis medicinal materials, peak 8 comes from Cuscuta chinensis medicinal materials, peak 9 comes from Rehmannia glutinosa medicinal materials, Eucommia ulmoides medicinal materials and Angelica dahurica medicinal materials, peak 10 comes from Myrrh medicinal materials and Cornus officinalis medicinal materials Cornus medicinal materials, peak No. 11 comes from Eucommia ulmoides medicinal materials, peak No. 12 comes from Cornus medicinal materials, peak No. 13 comes from Cuscuta australis medicinal materials, peak No. 14 comes from Cornus medicinal materials, peak No. 15 comes from Psoralea corylifolia medicinal materials, peak No. 16 comes from Psoralea corylifolia medicinal materials, peak No. 17 comes from Cuscuta australis medicinal materials, peak No. 18 comes from Cuscuta australis medicinal materials, peak No. 19 comes from Cuscuta australis medicinal materials, peak No. 20 comes from Cuscuta australis medicinal materials, peak No. 21 comes from Angelica sinensis medicinal materials, peak No. 22 comes from Angelica sinensis medicinal materials, peak No. 23 comes from Cuscuta australis medicinal materials, peak No. 24 comes from Psoralea corylifolia medicinal materials, and peak No. 25 comes from Psoralea corylifolia medicinal materials.

52. A method for detecting a Chinese medicinal compound containing Rehmannia glutinosa and Psoralea corylifolia, characterized in that: The detection method comprises the following steps: (1) Establishing a standard fingerprint of a Chinese medicine compound reference sample according to the fingerprint construction method described in any one of claims 1 to 51; (2) taking a test sample solution and testing it according to the chromatographic conditions in the fingerprint construction method according to any one of claims 1 to 51 to obtain a fingerprint of the Chinese medicine compound sample to be tested; as well as (3) Compare the fingerprint of the Chinese medicine compound sample obtained in step (2) with the standard fingerprint of the Chinese medicine compound reference sample obtained in step (1). If it meets the requirements, it is a qualified product, and if it does not meet the requirements, it is an unqualified product.

53. The detection method according to claim 52, characterized in that: The compliance requirements include one or more of the following: (1) The fingerprint of the Chinese medicine compound sample to be tested presents 25 characteristic chromatographic peaks, and the retention time of each characteristic chromatographic peak is within ±10% of the retention time value of the corresponding reference chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; (2) With peak No. 15 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine compound sample to be tested and the S peak is within ±10% of the relative retention time value of each characteristic chromatographic peak in the standard fingerprint of the Chinese medicine compound reference sample; as well as (3) According to the similarity evaluation system of Chinese medicine chromatographic fingerprint, the similarity between the fingerprint of the Chinese medicine compound sample to be tested and the standard fingerprint of the Chinese medicine compound reference sample shall not be less than 0.

90.

54. Use of the construction method according to any one of claims 1 to 51 or the detection method according to claim 52 or 53 in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine compound comprising Rehmannia glutinosa and Psoralea corylifolia.

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