Method and system for determining material unit representing traditional Chinese medicine and compound function characteristics, and readable storage medium

Abstract

The invention provides a method and system for determining a material unit representing function characteristics of a traditional Chinese medicine and a compound and a readable storage medium, and relates to the technical field of biological medicine.The method comprises the following steps that components of all medicinal materials in the traditional Chinese medicine and the compound are analyzed, and functions of the traditional Chinese medicine and the compound are determined; extracting according to the property of the traditional Chinese medicine component cluster, and establishing a fingerprint spectrum; determining individual'substance units' according to the fingerprint of the traditional Chinese medicine, determining structural'substance units' by adopting a matching frequency method, and determining functional'substance units' and administration'substance units' by adopting a factor analysis method; according to the method, the traditional Chinese medicine and the compound are taken as raw materials, the administration'material units' are comprehensively scored and sorted, functions are determined, a'material unit 'determination characterization system under traditional Chinese medicine and compound function characterization is formed, the process can be combined with various traditional Chinese medicines and compounds, research conditions of theories, production and application of biological medicines are included, and the method has wide market prospects and economic benefits.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and more specifically to a method, system and readable storage medium for determining material units that characterize the functional characteristics of traditional Chinese medicine and compound prescriptions. Background Art

[0002] Traditional Chinese medicine, compound prescriptions and their preparations (collectively referred to as products) have relatively prescribed processing methods and usage methods. They are preparations designed for relatively certain diseases and symptoms, and are the material basis for the clinical treatment of diseases with traditional Chinese medicine. However, the ingredients of traditional Chinese medicine, compound prescriptions and their preparations are complex and diverse, and the physical and chemical properties of each ingredient vary greatly. Traditional Chinese medicine fingerprints can characterize the single "substance unit" of the function of traditional Chinese medicine and compound prescriptions, but how to interpret the structure, function and drug administration "substance unit" from the complex fingerprint chromatographic peaks is a key issue in interpreting the theory of traditional Chinese medicine, and there is a lack of relevant research in the existing technology.

[0003] Therefore, how to provide a method to quickly determine and characterize the functional characteristics and size of a "material unit" is a core key technology for analyzing the original theories of traditional Chinese medicine and clarifying the basic laws of microscopic material operation. It has attracted great attention from the traditional Chinese medicine industry and is an issue that technical personnel in this field urgently need to solve. Summary of the invention

[0004] In view of this, the present invention provides a method, system and readable storage medium for determining material units that characterize the functional characteristics of traditional Chinese medicine and compound prescriptions, taking into account the autonomous action rules of the bio-supramolecular "imprinting template" of the current traditional Chinese medicine component clusters, and combining with the principles of multivariate statistics, it has the universality and flexibility of characterizing the effects of biological raw materials, controlling quality, and using them as material basic units for simplification, integration, and system characterization and correlation, and can be combined with various traditional Chinese medicines, including the theory, production and application research of biological drugs, and has broad market prospects and economic benefits.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] A method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription comprises the following steps:

[0007] S1: Analyze the literature and ingredients of each medicinal material in traditional Chinese medicine and compound prescriptions to determine the functions and material basis of traditional Chinese medicine and compound prescriptions;

[0008] S2: Extract the components of traditional Chinese medicine according to their properties and establish fingerprints;

[0009] S3: Determine the corresponding individual "substance unit" according to the fingerprint of traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method;

[0010] S4: Comprehensively score the drug administration "substance unit" and rank the scoring results. Determine the function of traditional Chinese medicine based on the ranking results, obtain the "substance unit" of the drug administration form under the functional characterization of traditional Chinese medicine and compound prescriptions, and establish a quantitative conversion relationship between individual "substance unit", structural "substance unit", functional "substance unit" and drug administration "substance unit".

[0011] Preferably, S3 includes:

[0012] S31: The component clusters of traditional Chinese medicine reflect the medicinal properties, efficacy and functions of traditional Chinese medicine in the form of supramolecular "imprinting templates" of traditional Chinese medicine;

[0013] S32: Determine individual “substance units” based on the fingerprint of traditional Chinese medicine;

[0014] S33: Integrate individual “material units” into structural “material unit” characteristics based on the supramolecular “imprinting template” of traditional Chinese medicine and the matching frequency main peak weight method.

[0015] Preferably, S3 further includes:

[0016] S34: Factor analysis and total variance analysis are used to integrate the structural "substance units" into functional "substance units", and the functional "substance units" are divided into monarch, minister, adjuvant and envoy levels to obtain the drug administration "substance units".

[0017] Preferably, the S34 further includes:

[0018] The rotating component matrix is ​​used to determine the constituent elements and relationships of the functional "material unit" and to determine the quantitative relationship between the structural "material unit" and the functional "material unit".

[0019] Preferably, S4 includes:

[0020] The scoring matrix comprehensive scoring method is used to calculate the factor contribution of the "substance unit" in each preparation, and quantitatively evaluate the effect of the "substance unit" in traditional Chinese medicine or compound on the function.

[0021] Preferably, the S33 includes:

[0022] Statistics are used to calculate the minimum frequency of the main chromatographic peak that can represent the structural "material unit" of the matching chromatographic peak, and then the retention time boundary and range between the two main peaks are divided according to the area size of the main chromatographic peak. The length of the range retention time is proportional to the area of ​​the chromatographic peak.

[0023] Preferably, the S33 includes:

[0024] The segment-band total statistical moment parameters are used to characterize the structural "material unit" characteristics of traditional Chinese medicine ingredient clusters.

[0025] The present invention also provides a system for determining material units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions, comprising:

[0026] The first analysis module is used to analyze the literature and ingredients of each medicinal material in the traditional Chinese medicine and compound prescriptions to determine the functions and material basis of the traditional Chinese medicine and compound prescriptions;

[0027] An extraction module is used to extract the components of traditional Chinese medicine according to their properties and to establish a fingerprint map;

[0028] The determination module is used to determine the corresponding individual "substance unit" according to the fingerprint of the traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method;

[0029] The second characterization module is used to comprehensively score the drug administration "material unit" and sort the scoring results, determine the function of the traditional Chinese medicine based on the sorting results, obtain the "material unit" of the drug administration form under the characterization of the function of traditional Chinese medicine and compound prescriptions, and establish a quantitative conversion relationship between individual "material unit", structural "material unit", functional "material unit" and drug administration "material unit".

[0030] The present invention also provides a computer-readable storage medium storing a computer program, which, when executed by a processor, implements the method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription as described in any one of the above items.

[0031] It can be seen from the above technical solutions that, compared with the prior art, the present invention discloses a method, system and readable storage medium for determining material units that characterize the functional characteristics of traditional Chinese medicine and compound prescriptions. Various "material units" under the functional characterization of traditional Chinese medicine and compound prescriptions can be quickly solved by dividing the frequency weights of fingerprint spectrum matching and combining factor analysis to establish an interrelated quantitative characterization system. The method provided by the present invention can solve the "bottleneck" problem of unclear operation of the material basis under functional characterization, and also has the following beneficial effects:

[0032] 1. The matching frequency main peak weight method used in the present invention can integrate the structural "substance unit" of the chromatographic peak of the traditional Chinese medicine fingerprint: because the traditional Chinese medicine and compound ingredients act on the biological network of the body according to the supramolecular "imprinting template", their overall effect is exerted. The supramolecular "imprinting template" is "a template that can be completely matched in terms of spatial structure and binding site", that is, "imprinting template" is a supramolecular chemical concept, which refers to the common molecular structural characteristics of a certain type of component, which can be understood as the spatial lattice of the effective atomic groups acting on the supramolecular object (traditional Chinese medicine component) and the human target, which is equivalent to the cooperation relationship between the teeth of the key and the lock. This relationship can be measured by chromatographic imprinting: Since the Chinese medicine component group is the result of the autonomous modification of the biological "imprint template", it presents the characteristics of the same parent nucleus group, which is reflected in the fingerprint spectrum. The chromatographic peak matching frequency will be distributed in a convex and concave manner with respect to the retention time, that is, an uneven cluster distribution. The closer the component is to the "imprint template", the greater the structural similarity, the higher the content, the larger the area, and the higher the frequency of occurrence. The frequency reflected in the matching of multiple sample fingerprints is greater. In this way, the main chromatographic peak of the structural "imprint template" can be determined according to the statistical principle of frequency. In the past, patents used the statistical moment principle of matching frequency to determine the number of two main chromatographic peaks. The boundary value of the two lines was calculated using the matching frequency total statistical moment method. The larger the peak area, the higher the matching frequency, and the closer the retention time. This is contradictory to the fact that the Chinese medicine or compound component cluster is superimposed according to the "imprint template", the larger the peak area, the greater the impact, and the wider the retention time range. Therefore, the present invention uses the main chromatographic peak weight method for correction.

[0033] 2. The factor analysis method used in the present invention can integrate the structural "substance units" of the chromatographic peaks of the traditional Chinese medicine fingerprint into functional "substance units": because each structural "substance unit" has the ability to perform certain biological functions. The more similar (or similar) biological functions are performed, or the traditional Chinese medicine or compound "substance units" are in the same metabolic pathway, the stronger the biological linkage, the better the correlation, the easier it is to be collinear, and the more it can be reflected in certain biological function factors. Therefore, through factor analysis, especially through factor rotation analysis, it is possible to find functional "substance units" that are independent of each other and can reflect their functional characteristics.

[0034] 3. The factor analysis method used in the present invention can integrate the effect size of the "substance unit" that characterizes the functions of various forms of Chinese medicine and compound prescriptions: Since the structural and functional "substance units" are intertwined and mixed in the original plants of Chinese medicine, Chinese medicine and compound prescriptions can be administered in various forms such as monomers, effective parts, decoction pieces, compound preparations, etc. Different forms of administration correspond to a group of functional "substance unit" factors, and the overall effect size can be calculated by comprehensive scoring based on the score coefficient matrix, and finally the effect size, quality, and advantages and disadvantages of the "substance unit" of Chinese medicine and compound prescriptions under certain functional characterization can be obtained.

[0035] In summary, the present invention takes into account the autonomous action rules of the bio-supramolecular "imprinting template" of the current traditional Chinese medicine ingredient clusters, and combines the principles of multivariate statistics. It has the universality and flexibility of characterizing the effects of biological raw materials, controlling their quality, and using them as the material basis unit for simplification, integration, and system characterization and association. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0037] Figure 1 The overall flow chart of a method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription provided by the present invention;

[0038] Figure 2 A structural principle block diagram of a system for determining a substance unit characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions provided by the present invention;

[0039] Figure 3 Schematic diagram of the main peak weighting method provided in Example 1 of the present invention, wherein t R,i ,t R,i+1 ,t R,(i～i+1) are the retention times of main chromatographic peak i, main chromatographic peak i+1, and boundary respectively; A i , A i+1 are the peak areas of the main chromatographic peaks, respectively;

[0040] Figure 4 The UPLC fingerprints of 13 yin-tonifying Chinese medicines and Liuwei Dihuang Decoction provided in Example 2 of the present invention, wherein S1 to S21 are Glehnia littoralis, Glehnia littoralis, Asparagus cochinchinensis, Lilium bulbiferum, Polygonatum odoratum, Polygonatum sibiricum, Sesame nigrum, Mulberry, Eclipta prostrata, Fructus Lycii, Fructus Ligustri Lucidi, Ophiopogon japonicus, Dendrobium officinale, Liuwei Dihuang Decoction, Morroniside, Loganin, Ferulic acid, Coumarin, Baicalin, Apigenin, and Methanol solvent blank, respectively. DETAILED DESCRIPTION

[0041] 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 only 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 ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0042] Example 1

[0043] See also Figure 1 As shown, Example 1 of the present invention discloses a method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription, comprising the following steps:

[0044] S1: Analyze the literature and ingredients of each medicinal material in traditional Chinese medicine and compound prescriptions, determine the functions and material basis of traditional Chinese medicine and compound prescriptions, obtain existing literature on traditional Chinese medicine through big data, including classic works, textbooks, monographs, papers, reports, and determine the clusters of traditional Chinese medicine and compound prescription ingredients with the same (similar) functions. The functions include the functions, functions, medicinal properties, efficacy, pharmacological effects, biological and chemical properties, main indications and indications of traditional Chinese medicine;

[0045] S2: Extract according to the properties of the Chinese medicine component clusters and establish a fingerprint spectrum, wherein the method for establishing the fingerprint spectrum can be to use high performance liquid chromatography (HPLC), ultra high performance liquid chromatography (UPLC), gas chromatography (GC), high performance liquid chromatography-mass spectrometry (HPLC-MS), gas chromatography-mass spectrometry (GC-MS), infrared spectroscopy (IR) and other optical and electromagnetic methods to establish the fingerprint spectrum of Chinese medicine;

[0046] S3: Determine the corresponding individual "substance unit" according to the fingerprint of traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method;

[0047] S4: Comprehensively score the drug administration "substance unit" and rank the scoring results. Determine the function of traditional Chinese medicine based on the ranking results, obtain the "substance unit" of the drug administration form under the functional characterization of traditional Chinese medicine and compound prescriptions, and establish a quantitative conversion relationship between individual "substance unit", structural "substance unit", functional "substance unit" and drug administration "substance unit".

[0048] Specifically, Chinese medicine and human body produce "gas separation" according to the supramolecular "imprinting template" of Chinese medicine, and four forms of "substance units" can be established based on this, namely individual, structural, functional and drug administration. The "substance unit" used to characterize the functions of Chinese medicine and compound prescriptions means that both Chinese medicine and human body are biological supramolecular bodies. In the biological evolution process in nature, living organisms synthesize biological supramolecular bodies on the basis of many small molecule templates through self-organization, self-assembly, self-recognition and self-replication. Its objects and human meridians and internal organs produce "gas separation" according to the "imprinting template", showing drug effects. The object molecule is an individual "substance unit" that can be quantitatively determined by fingerprint spectrum, reflecting characteristic and non-characteristic peaks; it can be integrated into structural "substance units" by matching frequency main peak weight method according to the "imprinting template"; it can be integrated into functional "substance units" by factor analysis; and the factor contribution degree of each Chinese medicine and compound drug administration "substance unit" can be calculated by comprehensive scoring of the scoring matrix. The four "substance units" are interconnected and quantitatively converted, and can be used for Chinese medicine theory and new drug research, Chinese medicine preparation and production quality evaluation.

[0049] In a specific embodiment, S3 includes:

[0050] S31: The component clusters of traditional Chinese medicine reflect the medicinal properties, efficacy and functions of traditional Chinese medicine in the form of supramolecular "imprinting templates" of traditional Chinese medicine;

[0051] S32: Determine individual “substance units” based on the fingerprint of traditional Chinese medicine;

[0052] S33: Integrate individual “material units” into structural “material units” based on the supramolecular “imprinting template” of traditional Chinese medicine and the matching frequency main peak weight method.

[0053] In a specific embodiment, S3 further includes:

[0054] S34: Factor analysis and total variance analysis are used to integrate the structural "substance units" into functional "substance units", and the functional "substance units" are divided into monarch, minister, adjuvant and envoy levels to obtain the drug administration "substance units".

[0055] In a specific embodiment, S34 further includes:

[0056] The rotating component matrix is ​​used to determine the constituent elements of the functional "material unit" and the quantitative relationship between the structural "material unit" and the functional "material unit".

[0057] Specifically, factor analysis is used to determine the monarch, minister, assistant and envoy levels of the functional "material units" composed of structural "material units" based on total variance analysis; determining the constituent elements and relationships of the functional "material units" based on the rotated component matrix refers to factor analysis of the structural "material units" to obtain the quantitative relationship with the structural "material units".

[0058] Factor analysis is a multivariate statistical method for dimensionality reduction. It is an extension and development of principal component analysis. It studies the internal dependencies of the correlation matrix or covariance matrix, explores the basic structure of the sample data set with minimal or no information loss, and integrates multiple variables into a few potential factors. These factors can highly summarize the information of a large number of samples and almost fully express the relationship between the original variables and the factors. It mainly includes the following steps:

[0059] Assume that there are P structural "substance units" in various drug delivery forms, which are regarded as variables and represented as k1, k2, k3, ..., k PX According to the principle of factor analysis, we first assume that these structural "material units" have been standardized (mean is 0, standard deviation is 1), and assume that P structural "material unit" variables can be represented by m functional factors as a linear combination, that is, the specific expression of the mathematical model of factor analysis in the form of a matrix is:

[0060] K p×1 =A p×m ·F m×1 +ε p×1 (1)

[0061] Where K is a measurable p-dimensional random vector, each of whose components represents a variable or indicator: F = (F1, F2, F3, ..., F m ) T is a common factor, the matrix A is called the factor loading matrix, and the vector ε is a special factor, which represents the part of the original variable that cannot be explained by the factor, with a mean of zero, including random errors.

[0062] Therefore, the first embodiment of the present invention can analyze the functional behavior of the structural "imprinting template" through factor analysis, and the factors with the same or similar functional "imprinting template" clusters are also put forward as the "behind-the-scenes" vertical manipulation factors, which are characterized as the combination of similar "substance units". This process is a linear transformation. Then, the eigenvalue size of each factor and the variance size are used to determine the monarch, minister, assistant, and envoy levels of each "substance unit" of the factor. After rotation matrix analysis, the factor loading matrix A p×m By comparing the sizes of the elements, we can know the composition and attribution of the structural "material unit", and realize the quantitative conversion between the structural "material unit" and the functional "imprinting template".

[0063] In a specific embodiment, S33 includes:

[0064] Statistics are used to calculate the minimum frequency of the main chromatographic peak that can represent the structural "material unit" of the matching chromatographic peak, and then the retention time boundary and range between the two main peaks are divided according to the area size of the main chromatographic peak. The retention time length of this range is proportional to the area of ​​the chromatographic peak.

[0065] In a specific embodiment, S33 further includes:

[0066] The structural "material unit" of the cluster of traditional Chinese medicine ingredients is characterized by the segmental total statistical moment parameters.

[0067] Specifically, the relationship between the retention times of the two main chromatographic peaks in S33 can be found in Figure 3 As shown, the specific expression is:

[0068]

[0069] In the formula, t R,i ,t R,i+1 ,t R,(i～i+1) are the retention times of main chromatographic peak i, main chromatographic peak i+1, and boundary respectively; A i , A i+1 are the peak areas of the main chromatographic peaks respectively.R,(i～i+1) If the retention time is the same as the boundary chromatogram, it will be uniformly classified into the "material unit" of the main chromatographic peak with a smaller retention time.

[0070] In S33, the structural "substance units" of Chinese medicine and compound ingredient clusters are divided by the pattern spectrum matching frequency method, which can be characterized by the total statistical moment parameters of the segment band, including:

[0071] (1) The zero-order moment of the total amount of the band is the total area of ​​the chromatographic peak within the defined range, reflecting the superposition of the overall content of the single and structural "substance units". The specific expression is:

[0072]

[0073] In the formula, AUC T,i , A j are the sum of the peak areas of the i-th “substance unit” component cluster and the peak area of ​​a single component; n i is the number of chromatographic peaks of the i-th "substance unit" component cluster.

[0074] (2) The first-order moment of the total amount of the segment band is the statistical average retention time of the chromatographic peak area within the defined range, reflecting the average relationship between the imprinting of a single component and the total amount. The specific expression is:

[0075]

[0076] In the formula, MCRT T,i , j are the average retention time of the i-th “substance unit” component cluster and the retention time of a single component peak, respectively.

[0077] (3) The second-order moment of the total amount of the segment band is the variance of the statistical average retention time of the chromatographic peak area within the defined range, which reflects the superposition relationship between the discrete degree of the imprinting of a single component and the discrete degree of the overall imprinting. The specific expression is:

[0078]

[0079] Where, VCRT T,i , They are the average retention time variance of the i-th “substance unit” component cluster and the retention time variance of a single component peak, respectively.

[0080] In a specific embodiment, S4 includes:

[0081] The factor contribution degree of each drug administration "substance unit" was calculated using the scoring matrix comprehensive scoring method.

[0082] Specifically, the function of Chinese medicine and compound prescriptions in different administration forms is comprehensively scored to determine their function size, and the "substance unit" under the function characterization of Chinese medicine and compound prescriptions in different administration forms is obtained by the score coefficient matrix obtained by factor analysis. The function of Chinese medicine and compound prescriptions in different administration forms is comprehensively scored and ranked to determine the function size. m×1 By multiplying the factor by the structural "material unit" in each dosage form and making a comprehensive score based on the size of the characteristic value, the overall contribution of the structural "material unit" in various dosage forms to the functional "material unit" factor can be calculated. Ultimately, the comprehensive score of the "material unit" in each dosage form can be determined to characterize the effect of various dosage forms on certain functions.

[0083] See also Figure 2 As shown, Embodiment 1 of the present invention further provides a system for a method of determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription using any one of the above embodiments, comprising:

[0084] The first analysis module is used to analyze the literature and ingredients of each medicinal material in traditional Chinese medicine and compound prescriptions to determine the functions and material basis of traditional Chinese medicine and compound prescriptions;

[0085] An extraction module is used to extract the components of traditional Chinese medicine according to their properties and to establish a fingerprint map;

[0086] The determination module is used to determine the corresponding individual "substance unit" according to the fingerprint of the traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method;

[0087] The second characterization module is used to comprehensively score the drug administration "material unit" and sort the scoring results, determine the function of the traditional Chinese medicine based on the sorting results, obtain the "material unit" of the drug administration form under the characterization of the function of traditional Chinese medicine and compound prescriptions, and establish a quantitative conversion relationship between individual "material unit", structural "material unit", functional "material unit" and drug administration "material unit".

[0088] Embodiment 1 of the present invention also provides a computer-readable storage medium, on which a computer program is stored. When the computer program is executed by a processor, the method for determining the material units that characterize the functional characteristics of traditional Chinese medicine and compound prescriptions as described in any of the above embodiments is implemented.

[0089] Example 2

[0090] The detailed process of the method provided in specific application example 1 is as follows:

[0091] This Example 2 selects Chinese medicine for tonifying yin and Liuwei Dihuang Decoction for structure, function and characterization of the "substance unit" of drug administration, which mainly includes the following steps:

[0092] 1. Source of medicinal materials and prescription

[0093] (1) Medicinal materials for tonifying yin: 13 Chinese medicinal materials, namely, Adenophora australis, Adenophora australis, Asparagus cochinchinensis, Lilium bulbus, Polygonatum odoratum, Polygonatum sibiricum, Sesame nigra, Mulberry, Ecliptae flavescentis, Fructus Lycii, Fructus Ligustri Lucidi, Ophiopogon japonicus, and Dendrobium candidum;

[0094] (2) Compound prescription for tonifying Yin: The prescription and preparation method of Liuwei Dihuang Decoction: 1) Prescription: 24g of Rehmannia glutinosa, 12g of Cornus officinalis, 9g of Paeonia suffruticosa, 12g of Dioscorea opposita, 9g of Poria cocos, and 9g of Alisma orientalis;

[0095] 2.13 Establishment of fingerprints of single Chinese medicine and Liuwei Dihuang Decoction

[0096] 2.1 Instruments and reagents

[0097] 2.1.1 Instruments: Ultra-high performance liquid chromatograph (Waters, USA, model: ACQUITY UPLC H-Class); rotary evaporator (Gongyi Zhongtian Instrument Technology Co., Ltd., model: RE-2000A); circulating water multi-purpose vacuum pump (Gongyi Zhongtian Instrument Technology Co., Ltd., model: SHB-III); electronic balance (Shimadzu Enterprise Management (China) Co., Ltd., model: AUY220); ultrasonic cleaner (Kunshan Hechuang Ultrasonic Instrument Co., Ltd., model: KH-300DE); temperature-adjustable electric heating mantle (Beijing Yongguangming Medical Instrument Co., Ltd., model: DZTW).

[0098] 2.1.2 Reagents: Acetonitrile (batch number: 22035335) and methanol (batch number: 22015112) were all chromatographically pure and purchased from Tiandi Reagent Company, USA; water was Yibao purified water (China Resources Yibao Beverage Co., Ltd., batch number: 14310Z10J); 0.22 μm microporous filter membrane (Tianjin Keyilong Experimental Equipment Co., Ltd., batch number: JLSN012201); Adenophora adenophora (batch number: 2310001), Ginseng (lot number: 2210001), Asparagus cochinchinensis (lot number: 231201), Lilium bulb (lot number: 231203), Polygonatum odoratum (lot number: 23080702), Polygonatum sibiricum (lot number: 231202), Black sesame (lot number: 22112001), Mulberry (lot number: 230901), Eclipta prostrata (lot number: 230301), Lycium barbarum (lot number: 231003), Ligustrum lucidum (lot number: 230901), Wheat Dong (batch number: 231001), Shihu (batch number: 231002), Rehmannia root (batch number: 231201), wine cornus (batch number: A231101), Moutan bark (batch number: 231001), Chinese yam (batch number: 231201), Poria (batch number: 231101), and Alisma orientalis (batch number: 2023020875) were purchased from Jiuzhitang Pharmacy near Hunan University of Traditional Chinese Medicine, and all of them comply with the requirements of the Chinese Pharmacopoeia 2020. The reference substances morroniside (DST230504-039), loganin (DST220901-038), coumarin (DX0013-0100), ferulic acid (DSTDF008102), baicalin (RM0523FA14), and apigenin (DSTDQ002602) were all purchased from Hunan Changsha Xiawei Biotechnology Co., Ltd., and the mass fraction of the above reference substances was greater than or equal to 98%.

[0099] 2.2 Methods and steps

[0100] 2.2.1 Preparation of test solution: Take about 20g of 13 yin-tonifying Chinese medicines such as single medicinal materials such as Adenophora australis, Adenophora australis, and Asparagus cochinchinensis, and Liuwei Dihuang Decoction (Rehmannia glutinosa: Cornus officinalis: Chinese yam: Paeonia suffruticosa: Poria: Alisma orientalis = 6.4:3.2:3.2:2.4:2.4:2.4) and add them to a round-bottom flask, add 8 times the amount of water and reflux and extract twice, each time for 1 hour, filter through a cloth filter bag, combine the two filtrates, and concentrate under reduced pressure to 1.5g of original medicinal materials / ml. Then accurately and easily take 5ml of the concentrate, add an appropriate amount of 80% methanol solution, ultrasonically extract (power 200w, frequency 40KHz) for 30min, filter, transfer to a 25ml volumetric flask, and then dilute to the mark with 80% methanol, pass through a 0.22μm organic filter membrane, take the subsequent filtrate, and place it in a sample injection vial for testing.

[0101] 2.2.2 Preparation of reference solution: Take appropriate amount of morroniside, loganin, coumarin, ferulic acid, baicalin and apigenin reference substances, accurately weigh 2 mg, place in a brown volumetric flask, add 80% methanol to make up to 10 ml, make 0.2 mg / ml reference solution, and ultrasonically treat for 5 min, pass through 0.22 μm organic filter membrane, place in injection vial for testing.

[0102] 2.2.3 Chromatographic determination conditions: the chromatographic column is Waters AcquityUPLC BEHC18 (2.1mm×100mm, 1.7μm); mobile phase: acetonitrile (B)-0.1% phosphoric acid water (A), gradient elution: 0-0.5min, 99.9%A; 0.5-20.5min, 99.9%-82.0%A; 20.5-23.5min, 82%-79%A; 23.5-30.5min, 79%-65%A; 30.5-34.5min, 65%-55%A; 34.5-36.5min, 55%-32%A; 36.5-37.5min, 32%-0%A; column temperature is 35°C; flow rate is 0.2mL / min; injection volume is 2μL; full wavelength scan is 200-400, and the legend shows that the selected wavelength is 250nm.

[0103] 2.2.4 Precision, stability and repeatability test

[0104] (1) Precision test: Take the same test solution prepared under "2.2.1" and inject it 6 times continuously according to the chromatographic conditions of "2.2.3". Record the fingerprint spectrum and the RSD of the relative peak area of ​​the reference substance morroniside is 3.46%, indicating that the precision of the experimental instrument is good.

[0105] (2) Stability test: The same test solution prepared under "2.2.1" was sampled and measured according to the chromatographic conditions of "2.2.3" at 2, 4, 6, 8 and 12 hours after preparation. The fingerprint was recorded and the RSD of the relative peak area of ​​the reference substance morroniside was 4.80%, indicating that the test solution had good stability within 12 hours.

[0106] (3) Repeatability test: Take the same batch of medicinal materials and operate according to the method in "2.2.1". Prepare 6 test sample solutions in parallel. Inject and measure according to the chromatographic conditions in "2.2.3". Record the fingerprint spectrum. The relative peak area RSD of the reference substance morroniside is 6.20%, indicating that the method has good repeatability.

[0107] 3 Results

[0108] 3.1 Establishment of UPLC fingerprint: Take the sample solution prepared under "2.2.1" and inject it, and measure the UPLC fingerprint superposition of 13 Chinese medicines including Glehnia littoralis, Glehnia littoralis, Asparagus cochinchinensis, Lilium bulbus, Polygonatum odoratum, Polygonatum sibiricum, Sesame nigrum, Morus alba, Ecliptae macrophylla, Fructus Lycii, Fructus Ligustri Lucidi, Ophiopogon japonicus, Dendrobium officinale and Liuwei Dihuang Decoction. See for details. Figure 4 .

[0109] 3.2 Individual “substance units” and total statistical moment parameters of UPLC fingerprint

[0110] According to the total statistical moment parameter formula, the peak number, total zero-order moment, first-order moment, and second-order moment parameter values ​​of 13 Chinese medicinal materials including Glehnia littoralis, Glehnia littoralis, Asparagus cochinchinensis, Lilium bulbiferum, Polygonatum odoratum, Polygonatum sibiricum, Sesame seed, Morus alba, Ecliptae macrophylla, Fructus Lycii, Fructus Ligustri Lucidi, Ophiopogon japonicus, and Dendrobium officinale and Liuwei Dihuang Decoction were obtained, and the values ​​are listed in the corresponding rows of the state "front" in Table 1.

[0111] Table 1 Comparison of the total statistical moment parameter characteristics of individual and structural “substance units” of HPLC fingerprints of 13 yin-tonifying Chinese medicines and Liuwei Dihuang Decoction

[0112]

[0113]

[0114] 3.3 Structural “substance unit” division and total amount statistical moment parameters of UPLC fingerprint

[0115] According to the matching frequency main peak weight method mentioned above, the structural "substance units" of 13 yin-tonifying Chinese medicines and Liuwei Dihuang Decoction can be delineated, and the values ​​of the corresponding rows of the "after" state in Table 1 are listed. By comparing the "before" and "after" states, it can be seen that the relationship between the qualitative and quantitative transformation of individual "substance units" into structural "substance units" can be known. As shown in Table 1, the total statistical moment parameters of the "substance units" before and after integration are equal, that is, the imprinting of the overall material basis of the overall yin-tonifying medicine has not changed. The individual "substance units" can be represented by the compressed structural "substance units", and their quantitative relationship is determined by the segment total statistical moment formula.

[0116] 3.4 Analysis of the functional “substance unit” factors of UPLC fingerprint

[0117] The structure “substance unit” in Table 1 was subjected to factorial analysis, and the total variance analysis of its main factors is shown in Table 2.

[0118] Table 2. Total variance analysis of main factors of the functional “imprinting template” of Yin-tonifying Chinese medicine and Liuwei Dihuang Decoction

[0119]

[0120]

[0121] As shown in Table 2, the 23 structural "substance units" of traditional Chinese medicines with yin-tonifying function and Liuwei Dihuang Decoction can be integrated into no more than 13 functional "substance units". Among them, the first three functional "substance units" account for two-thirds of the entire yin-tonifying material basis. The composition of the elements (structural "substance units") of each functional "substance unit" is shown in Table 3.

[0122] As shown in Table 3, the first functional “substance unit” is composed of 9 structural “substance units”, whose retention times are 19.697, 20.577, 21.091, 19.486, 22.178, 19.970, 16.752, 24.397, and 11.603 min, respectively, and the corresponding contribution coefficients are 0.9882, 0.9870, 0.9711, 0.9707, 0.9389, and 0.912, respectively. 0, 0.9000, 0.8736, 0.7834, which concentrates the middle component cluster of the chromatogram and can be defined as the main component cluster; the second functional "substance unit" is composed of three structural "substance units", whose retention times are 32.231, 25.938, 27.147min, respectively, and the corresponding contribution coefficients are 0.9814, 0.9539, 0.7197, respectively; the third functional "substance unit" is composed of four structural "substance units". , and their retention times were 9.852, 29.977, 28.271, and 26.651 min, respectively, and the corresponding contribution coefficients were 0.9843, 0.7311, 0.6907, and 0.6882, respectively; the 4th functional "substance unit" was composed of 2 structural "substance units", and their retention times were 8.104 and 6.321 min, respectively, and the corresponding contribution coefficients were 0.9552 and 0.8184, respectively; the 5th functional "substance unit" was composed of 2 structural "substance units", and their retention times were 1.416 and 2.939 min, respectively, and the corresponding contribution coefficients were 0.9552 and 0.8184, respectively; the 6th, 7th, and 8th functional "substance units" were all composed of 1 structural "substance unit", and their retention times were 4.478, 36.942, and 2.081 min, respectively, and the corresponding contribution coefficients were 0.9766, 0.7110, and 0.7053, respectively. The factor contribution of these 8 yin-replenishing Chinese medicines is up to 98.89%, while other factors only account for 1.11% and can be ignored.

[0123] Table 3 Relationship matrix between the functional “material unit” and structural “material unit” of the rotation component

[0124]

[0125]

[0126] 3.5 Comprehensive factor scoring analysis of each Chinese medicine “substance unit” for tonifying Yin

[0127] The 13 yin-tonifying Chinese medicines and Liuwei Dihuang Decoction were normalized and comprehensively scored to obtain the comprehensive scores of each yin-tonifying Chinese medicine, as shown in Table 4.

[0128] As can be seen from Table 4, among the “material units” of the 13 Chinese herbs with yin-tonifying function and Liuwei Dihuang Decoction, mulberry has the highest and Ophiopogon japonicus has the lowest, but Liuwei Dihuang Decoction can be ranked in the top 5.

[0129] Table 4 Comprehensive score ranking results of Yin-tonifying function of each Yin-tonifying Chinese medicine (Liuwei Dihuang Pills) in the administration of “substance unit”

[0130]

[0131] In summary, the qualitative and quantitative measurement of the individual, structure, function and administration "material unit" of the Yin-tonifying Chinese medicine (Liuwei Dihuang Pills) achieved in Example 2 illustrates the relationship between the qualitative and quantitative conversion of various forms of "material units". If each "material unit" is further analyzed in association with a certain pharmacodynamics, it will be possible to achieve quantitative analysis from microscopic component clusters, effector molecules, targets to macroscopic functional effects, illustrate the various functions of Chinese medicine and compound prescriptions, and realize modernization.

[0132] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the embodiments can be referred to each other. As for the contents disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple, and the relevant parts can be referred to the method part.

[0133] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine or a compound prescription, characterized in that: The following steps are involved: S1: Analyze the literature and ingredients of each medicinal material in traditional Chinese medicine and compound prescriptions to determine the functions and material basis of traditional Chinese medicine and compound prescriptions; S2: Extract the components of traditional Chinese medicine according to their properties and establish fingerprints; S3: Determine the corresponding individual "substance unit" according to the fingerprint of traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method; S4: Comprehensively score the drug administration "substance unit" and rank the scoring results. Determine the function of traditional Chinese medicine based on the ranking results, obtain the "substance unit" of the drug administration form under the functional characterization of traditional Chinese medicine and compound prescriptions, and establish the quantitative conversion relationship between individual "substance unit", structural "substance unit", functional "substance unit" and drug administration "substance unit".

2. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 1, characterized in that: The S3 includes: S31: The component clusters of traditional Chinese medicines reflect the properties, efficacy and functions of traditional Chinese medicines in the form of supramolecular "imprinting templates" of traditional Chinese medicines; S32: Determine individual "substance units" based on the fingerprint of traditional Chinese medicine; S33: Integrate individual "substance units" into structural "substance units" based on the supramolecular "imprinting template" of traditional Chinese medicine and the matching frequency main peak weight method.

3. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 2, characterized in that: The S3 further includes: S34: Factor analysis and total variance analysis were used to integrate the structural "substance units" into functional "substance units", and the functional "substance units" were divided into monarch, minister, adjuvant and envoy levels to obtain the drug administration "substance units".

4. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 2, characterized in that: The S34 further includes: The rotation component matrix is ​​used to determine the constituent elements and relationships of the functional "material unit" and to determine the quantitative relationship between the structural "material unit" and the functional "material unit".

5. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 1, characterized in that: The S4 includes: The scoring matrix comprehensive scoring method was used to calculate the factor contribution of the "substance unit" in each preparation, and to quantitatively evaluate the effect of the "substance unit" in traditional Chinese medicine or compound on the function.

6. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 2, characterized in that: The S33 includes: Statistics are used to calculate the minimum frequency of the main chromatographic peak that can represent the structural "material unit" of the matching chromatographic peak, and then the retention time boundary and range between the two main peaks are divided according to the area size of the main chromatographic peak. The length of the range retention time is proportional to the area of ​​the chromatographic peak.

7. A method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to claim 2, characterized in that: The S33 includes: The structural "material unit" characteristics of the clusters of traditional Chinese medicine components are characterized by the segmental total statistical moment parameters.

8. A system using the method for determining substance units characterizing the functional characteristics of traditional Chinese medicine and compound prescriptions according to any one of claims 1 to 7, characterized in that: include: The first analysis module is used to analyze the literature and ingredients of each medicinal material in the traditional Chinese medicine and compound prescriptions to determine the functions and material basis of the traditional Chinese medicine and compound prescriptions; An extraction module is used to extract the components of traditional Chinese medicine according to their properties and to establish a fingerprint map; A determination module is used to determine the corresponding individual "substance unit" according to the fingerprint of the traditional Chinese medicine; determine the structural "substance unit" according to the matching frequency method; determine the functional "substance unit" and the drug administration "substance unit" according to the factor analysis method; The second characterization module is used to comprehensively score the drug administration "substance unit" and sort the scoring results, determine the function of traditional Chinese medicine according to the sorting results, obtain the "substance unit" of the drug administration form under the function characterization of traditional Chinese medicine and compound prescriptions, and establish the quantitative conversion relationship between individual "substance unit", structural "substance unit", functional "substance unit" and drug administration "substance unit".

9. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the method for determining a substance unit characterizing the functional characteristics of a traditional Chinese medicine and a compound prescription according to any one of claims 1 to 7 is implemented.

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