Fingerprint spectrum construction method and application of traditional Chinese medicine composition containing angelica sinensis and cinnamon

Abstract

The invention discloses a fingerprint spectrum construction method of a traditional Chinese medicine composition containing angelica sinensis and cinnamon. The construction method comprises the following steps: preparing a test solution; preparing a reference substance solution; and obtaining the fingerprint spectrum of the traditional Chinese medicine composition according to results of high performance liquid detection of the test solution and the reference solution. The chemical components of the traditional Chinese medicine composition containing angelica sinensis and cinnamon are comprehensively and systematically analyzed, and a theoretical basis is provided for deep research on quality control and pharmacodynamic material basis.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a fingerprint construction method of a traditional Chinese medicine composition containing angelica and cinnamon bark 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 of the chemical components and fingerprint study of the Chinese medicine composition comprising angelica and cinnamon claimed in the present invention. In the prior art, only a single component in the 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 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 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 method for constructing a fingerprint of a traditional Chinese medicine composition containing angelica and cinnamon, and the construction method comprises the following steps:

[0007] Preparation of a Chinese medicine composition test solution: taking an appropriate amount of a decoction of a Chinese medicine composition containing angelica and cinnamon, placing it in a container, adding a solvent, ultrasonically treating it for a period of time, fixing the volume, shaking it well, centrifuging it, filtering the supernatant, and taking the filtrate to obtain the Chinese medicine composition test solution; wherein the Chinese medicine composition includes angelica, cinnamon, white peony root, ginger, jujube and liquorice;

[0008] Preparation of reference solution: Weigh appropriate amounts of gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide reference substances, add methanol to prepare reference solutions with concentrations of 1 to 100 μg / ml for gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide, respectively;

[0009] Obtaining a fingerprint of the Chinese medicine composition according to the results of high performance liquid chromatography detection of the test solution and the reference solution of the Chinese medicine composition;

[0010] The chromatographic conditions of the high performance liquid phase detection are as follows: a chromatographic column with octadecyl bonded silica as filler is used, mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, mobile phase B is an acid aqueous solution, an alkali aqueous solution and / or a buffered saline solution, and the gradient elution program is as follows: 0-10 min, 95%→94% B; 10-12 min, 94% B→88% B; 12-20 min, 88% B; 20-21 min, 88%→84% B; 21-38 min, 84% B; 38-48 min, 84% B→80% B; 48-52 min, 84% B→80% B; 48-52 min, 84% B→80% B; 48-52 min, 84% B→80% B. min, 80%→75% B; 52~58min, 75% B→71% B; 58~62min, 71%→68% B; 62~80min, 68% B→44% B; 80~83min, 44% B→26% B; 83~88min, 26% B→15% B; 88~89min, 15% B→95% B; 89~100min, 95% B; the flow rate is 0.5~1.5ml / min, the column temperature is 10~40℃, the detection wavelength is 180~350nm, and the injection volume is 1~20μl.

[0011] Furthermore, the licorice is stir-fried licorice.

[0012] Furthermore, in the preparation of the test solution of the traditional Chinese medicine composition, the solvent is an alcohol, such as methanol.

[0013] Furthermore, the container is a volumetric flask.

[0014] Furthermore, the mass of the decoction of the traditional Chinese medicine composition is 5 to 30 g, for example, about 15 g.

[0015] Furthermore, the mass-to-volume ratio between the decoction of the traditional Chinese medicine composition and the solvent is 0.4-0.8, such as about 0.6, in units of g / ml.

[0016] Furthermore, the power of the ultrasonic treatment is 400-600W, for example, about 500W.

[0017] Furthermore, the frequency of the ultrasonic treatment is 30-50 kHz, for example, about 40 kHz.

[0018] Furthermore, the ultrasonic treatment time is 20 to 40 minutes, for example, about 30 minutes.

[0019] Furthermore, the centrifugal speed is 4000-6000 rpm / min, for example, about 5000 rpm / min.

[0020] Furthermore, the centrifugation time is 5 to 15 minutes, for example, about 10 minutes.

[0021] Furthermore, the mass ratios of the angelica, cinnamon bark, white peony root, ginger, jujube and liquorice are (45-60):(30-50):(70-90):(30-50):(25-45):(20-40).

[0022] Furthermore, the mass ratio of the angelica sinensis, the cinnamon bark, the white peony root, the ginger, the jujube and the licorice is about 55.2: about 41.4: about 82.8: about 41.4: about 36: about 27.6.

[0023] Furthermore, the preparation method of the decoction of the Chinese medicine composition containing angelica and cinnamon includes: weighing appropriate amounts of angelica, cinnamon, white peony root, ginger, jujube and licorice, crushing them into coarse particles, placing them in a container, soaking them in water, decocting them until boiling and boiling them for a period of time, then keeping them slightly boiling for a period of time, filtering the filtrate with a filter while it is hot, and obtaining the decoction of the Chinese medicine composition containing angelica and cinnamon.

[0024] Furthermore, the volume / mass ratio of the water to the traditional Chinese medicine composition comprising angelica and cinnamon is 5-10, for example, about 7.

[0025] Furthermore, the container is a ceramic medicine decoction pot or an electric ceramic stove.

[0026] Furthermore, the soaking time is 10 to 100 minutes, for example, about 45 minutes.

[0027] Furthermore, the decoction is covered when it boils.

[0028] Furthermore, the power of the container during boiling is 1000-3000W, for example, about 2000W.

[0029] Furthermore, the boiling time is 5 to 15 minutes, for example, about 10 minutes.

[0030] Furthermore, the power of the container when slightly boiling is 500-1500W, for example, about 900W.

[0031] Furthermore, the slight boiling time is 10 to 100 minutes, for example, about 60 minutes.

[0032] Furthermore, the volume / volume ratio of the decoction of the Chinese medicinal composition comprising angelica and cinnamon to the water is 0.1 to 0.5, such as about 0.3.

[0033] Furthermore, the filter is a 200-mesh nylon filter.

[0034] Furthermore, the concentration of ferulic acid in the reference solution is 10-25 μg / ml, for example, about 18 μg / ml.

[0035] Furthermore, the concentration of cinnamaldehyde in the reference solution is 10-20 μg / ml, for example, about 13 μg / ml.

[0036] Furthermore, the concentration of 6-gingerol in the reference solution is 1-10 μg / ml, for example, about 6 μg / ml.

[0037] Furthermore, the concentration of ligustilide in the reference solution is 10-25 μg / ml, for example, about 18 μg / ml.

[0038] Furthermore, the gallic acid in the reference solution is 20-40 μg / ml, for example, about 30 μg / ml.

[0039] Furthermore, the concentration of paeoniflorin in the reference solution is 50-70 μg / ml, for example, about 60 μg / ml.

[0040] Furthermore, the cinnamic acid in the reference solution is 40-60 μg / ml, for example, about 50 μg / ml.

[0041] Furthermore, the glycyrrhizic acid in the reference solution is 40-60 μg / ml, for example, about 50 μg / ml.

[0042] Furthermore, the flow rate of the HPLC detection is 0.8-1.2 ml / min, for example, about 1.0 ml / min.

[0043] Furthermore, the column temperature is 20-30°C, for example, about 25°C.

[0044] Further, between the 0th minute and the 77th minute, the detection wavelength is 210-280 nm, for example, about 250 nm.

[0045] Further, between the 77th minute and the 100th minute, the detection wavelength is 190-230 nm, for example, about 210 nm.

[0046] Furthermore, the injection volume is 5-15 μl, for example, about 10 μl.

[0047] Furthermore, the theoretical plate number of the chromatographic peak corresponding to the ferulic acid is not less than 5000.

[0048] Furthermore, the chromatographic column is an Agilent 5TC-C18 chromatographic column.

[0049] Furthermore, the specifications of the chromatographic column are: column length 250 mm, inner diameter 4.6 mm, and particle size 5 μm.

[0050] Furthermore, the mobile phase A is acetonitrile.

[0051] Furthermore, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution is selected from one or more of weak acids and their salts, weak bases and their salts of different concentrations.

[0052] Further, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution is 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 in different concentrations.

[0053] Furthermore, the acid aqueous solution is a 0.01% to 1% acid aqueous solution.

[0054] Furthermore, the acid aqueous solution is a 0.01% to 1% phosphoric acid aqueous solution.

[0055] Furthermore, the acid aqueous solution is about 0.1% phosphoric acid aqueous solution.

[0056] Furthermore, the buffered saline solution is an acetate solution and / or an acetate solution.

[0057] Further, the pH value of the buffered saline solution is not greater than 7.0.

[0058] Further, when the detection wavelength between the 0th minute and the 77th minute is 250nm and the detection wavelength between the 77th minute and the 100th minute is 210nm, the fingerprint spectrum includes peaks 1-14, and the retention times of peaks 1-14 correspond to about 8.968min, about 21.263min, about 27.481min, about 29.945min, about 38.938min, about 42.241min, about 45.390min, about 64.056min, about 65.959min, about 69.988min, about 72.691min, about 74.593min, about 78.837min, and about 85.261min, respectively.

[0059] Further, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, the fingerprint spectrum includes peaks 1-14, wherein peak 4 is used as a reference peak, and the relative retention time average values ​​of peak 1, peak 2, peak 3, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, and peak 14 are approximately 0.299, approximately 0.710, approximately 0.918, approximately 1.300, approximately 1.411, approximately 1.516, approximately 2.139, approximately 2.203, approximately 2.337, approximately 2.427, approximately 2.491, approximately 2.633, and approximately 2.847, respectively.

[0060] Furthermore, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, peak 1 is gallic acid, peak 4 is paeoniflorin, peak 5 is ferulic acid, peak 8 is cinnamic acid, peak 9 is cinnamaldehyde, peak 12 is glycyrrhizic acid, peak 13 is 6-gingerol, and peak 14 is ligustilide.

[0061] Further, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, peak No. 1 comes from white peony medicinal material, peak No. 2 comes from white peony medicinal material, peak No. 3 comes from white peony medicinal material, peak No. 4 comes from white peony medicinal material, peak No. 5 comes from angelica medicinal material, peak No. 6 comes from stir-fried licorice medicinal material, peak No. 7 comes from white peony medicinal material, peak No. 8 comes from cinnamon medicinal material, peak No. 9 comes from cinnamon medicinal material, peak No. 10 comes from stir-fried licorice medicinal material, peak No. 11 comes from stir-fried licorice medicinal material, peak No. 12 comes from stir-fried licorice medicinal material, peak No. 13 comes from ginger medicinal material, and peak No. 14 comes from angelica medicinal material.

[0062] According to another aspect of the present invention, a quality control method for a Chinese medicine composition containing angelica and cinnamon is provided, the quality control method comprising the following steps:

[0063] (1) establishing a standard fingerprint of a reference sample of a Chinese medicine composition according to the above-mentioned fingerprint construction method;

[0064] (2) taking a sample solution of the Chinese medicine composition and testing it according to the chromatographic conditions in the fingerprint spectrum construction method to obtain a fingerprint spectrum of the Chinese medicine composition sample to be tested; and

[0065] (3) Compare the fingerprint of the sample of the Chinese medicine composition obtained in step (2) with the standard fingerprint of the reference sample of the Chinese medicine composition obtained in step (1). If the fingerprint meets the requirements, it is a qualified product; if the fingerprint does not meet the requirements, it is an unqualified product.

[0066] Furthermore, the compliance requirement includes one or more of the following:

[0067] (1) The fingerprint of the Chinese medicine composition sample to be tested exhibits 14 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 composition reference sample;

[0068] (2) With peak No. 4 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine composition 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 composition reference sample; and

[0069] (3) According to the similarity evaluation system of Chinese medicine chromatographic fingerprint, the similarity between the fingerprint of the Chinese medicine composition sample to be tested and the standard fingerprint of the Chinese medicine composition reference sample shall be calculated, and the similarity shall not be less than 0.90.

[0070] According to another aspect of the present invention, there is provided a use of the above construction method or the above quality control method in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine composition containing angelica and cinnamon.

[0071] Beneficial effects of the present invention:

[0072] Beneficial effects of the present invention:

[0073] In short, the present invention provides a fingerprint spectrum determination and quality control method for a reference sample of a traditional Chinese medicine composition containing angelica and cinnamon. The method confirms 14 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 stability of product quality, guarantees the efficacy of the traditional Chinese medicine compound, and allows the traditional Chinese medicine compound to better serve human life and health.

[0074] Specifically, compared with the prior art, the present invention has the following beneficial effects:

[0075] (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, using the main means of modern drug research, realizing the inheritance of classics, and making the classic prescriptions more formally controlled in quality;

[0076] (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 spectrum, the present invention adopts a gradient elution method to solve the problem of difficulty in separating the fingerprint characteristic peaks and interference of impurity peaks;

[0077] (3) In the process of establishing the fingerprint spectrum of the traditional Chinese medicine compound reference sample of the present invention, 14 common characteristic peaks were confirmed, and their relative retention time, relative peak area and similarity were studied, thereby ensuring the chemical composition stability and safety of the reference sample, and providing an important reference basis and quality reference for the quality control of the subsequent compound preparation;

[0078] (4) The fingerprints of the effective components in the traditional Chinese medicine compound reference sample of the present invention are viewed as a whole, and the order and relationship of each characteristic peak are emphasized, which not only avoids the one-sidedness of judging the overall quality of the traditional Chinese medicine compound reference sample of the present invention by only measuring one or two chemical components, but also reduces the possibility of artificial processing for the purpose of meeting the quality standard, thus providing a new method and means for completely and accurately evaluating the quality of the traditional Chinese medicine compound reference sample of the present invention;

[0079] (5) The method of the present invention has good stability, high precision, good reproducibility, is convenient and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] 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.

[0081] Figure 1 It is the liquid phase fingerprint of the reference sample of the Chinese medicine composition granules containing angelica and cinnamon of the present invention. DETAILED DESCRIPTION

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] As described in the background technology section, there is no literature report on a comprehensive analysis of the chemical components and fingerprint study of the Chinese medicine composition comprising angelica and cinnamon claimed in the present invention. In order to solve the above problem, the present invention provides a method for constructing a fingerprint of a Chinese medicine composition comprising angelica and cinnamon, and the construction method comprises the following steps:

[0088] Preparation of a Chinese medicine composition test solution: taking an appropriate amount of a decoction of a Chinese medicine composition containing angelica and cinnamon, placing it in a container, adding a solvent, ultrasonically treating it for a period of time, fixing the volume, shaking it well, centrifuging it, filtering the supernatant, and taking the filtrate to obtain the Chinese medicine composition test solution; wherein the Chinese medicine composition includes angelica, cinnamon, white peony root, ginger, jujube and liquorice;

[0089] Preparation of reference solution: Weigh appropriate amounts of gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide reference substances, add methanol to prepare reference solutions with concentrations of 1 to 100 μg / ml for gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide, respectively;

[0090] Obtaining a fingerprint of the Chinese medicine composition according to the results of high performance liquid chromatography detection of the test solution and the reference solution of the Chinese medicine composition;

[0091] The chromatographic conditions of the high performance liquid phase detection are as follows: a chromatographic column with octadecyl bonded silica as filler is used, mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, mobile phase B is an acid aqueous solution, an alkali aqueous solution and / or a buffered saline solution, and the gradient elution program is as follows: 0-10 min, 95%→94% B; 10-12 min, 94% B→88% B; 12-20 min, 88% B; 20-21 min, 88%→84% B; 21-38 min, 84% B; 38-48 min, 84% B→80% B; 48-52 min, 84% B→80% B; 48-52 min, 84% B→80% B; 48-52 min, 84% B→80% B. min, 80%→75% B; 52~58min, 75% B→71% B; 58~62min, 71%→68% B; 62~80min, 68% B→44% B; 80~83min, 44% B→26% B; 83~88min, 26% B→15% B; 88~89min, 15% B→95% B; 89~100min, 95% B; the flow rate is 0.5~1.5ml / min, the column temperature is 10~40℃, the detection wavelength is 180~350nm, and the injection volume is 1~20μl.

[0092] In the present invention, when concentration, time, flow rate, column temperature, wavelength, injection volume, ratio, power, frequency, mass, rotation speed, 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 with any lower range limit or preferred value, regardless of whether the range is disclosed separately. For example, when the range "0.5-1.5" is disclosed, the described range should be interpreted as including the ranges "0.5-1.5", "0.5-1.3", "0.5-1.1", "0.5-0.9", "0.5-0.7", "0.7-1.5", "0.7-1.3", "0.7-1.1", "0.7-0.9", "0.9-1.5", "0.9-1.3", "0.9-1.1", "1.1-1.5", "1.1-1.3", "1.3-1.5", 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.

[0093] In a preferred embodiment, the licorice is stir-fried licorice.

[0094] In a preferred embodiment, in the preparation of the test solution of the traditional Chinese medicine composition, the solvent is alcohol, such as methanol.

[0095] In a preferred embodiment, the container is a volumetric flask.

[0096] In a preferred embodiment, the mass of the decoction of the traditional Chinese medicine composition is 5 to 30 g, for example, about 15 g.

[0097] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 15" includes ±5% of 15, or from 14.25 to 15.75.

[0098] In a preferred embodiment, the mass-to-volume ratio between the decoction of the traditional Chinese medicine composition and the solvent is 0.4-0.8, such as about 0.6, in units of g / ml.

[0099] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.6" includes ±5% of 0.6, or from 0.57 to 0.63.

[0100] In a preferred embodiment, the power of the ultrasonic treatment is 400-600W, such as about 500W.

[0101] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 500" includes ±5% of 500, or from 475 to 525.

[0102] In a preferred embodiment, the frequency of the ultrasonic treatment is 30 to 50 kHz, such as about 40 kHz.

[0103] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 40" includes ±5% of 40, or from 38 to 42.

[0104] In a preferred embodiment, the ultrasonic treatment time is 20 to 40 minutes, such as about 30 minutes.

[0105] 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.

[0106] In a preferred embodiment, the centrifugal rotation speed is 4000-6000 rpm / min, such as about 5000 rpm / min.

[0107] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 5000" includes ±5% of 5000, or from 4750 to 5250.

[0108] In a preferred embodiment, the centrifugation time is 5 to 15 min, such as about 10 min.

[0109] 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.

[0110] In a preferred embodiment, the mass ratio of the angelica, cinnamon bark, white peony root, ginger, jujube and liquorice is (45-60):(30-50):(70-90):(30-50):(25-45):(20-40).

[0111] In a preferred embodiment, the mass ratio of the angelica sinensis, the cinnamon bark, the white peony root, the ginger, the jujube and the liquorice is about 55.2: about 41.4: about 82.8: about 41.4: about 36: about 27.6.

[0112] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 55.2" includes ±5% of 55.2, or from 52.44 to 57.96; "about 41.4" includes ±5% of 41.4, or from 39.33 to 43.47; "about 82.8" includes ±5% of 82.8, or from 78.66 to 86.94; "about 36" includes ±5% of 36, or from 34.2 to 37.8; "about 27.6" includes ±5% of 27.6, or from 26.22 to 28.98.

[0113] In a preferred embodiment, the preparation method of the decoction of the Chinese medicine composition containing angelica and cinnamon comprises: weighing appropriate amounts of angelica, cinnamon, white peony root, ginger, jujube and licorice, crushing them into coarse particles, placing them in a container, soaking them in water, decocting them until boiling and boiling them for a period of time, then keeping them slightly boiling for a period of time, filtering the filtrate with a filter while it is hot, and obtaining the decoction of the Chinese medicine composition containing angelica and cinnamon.

[0114] In a preferred embodiment, the volume / mass ratio of the water to the traditional Chinese medicine composition comprising angelica and cinnamon is 5-10, such as about 7.

[0115] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 7" includes ±5% of 7, or from 6.65 to 7.35.

[0116] In a preferred embodiment, the container is a ceramic medicine decoction pot or an electric ceramic stove.

[0117] In a preferred embodiment, the soaking time is 10 to 100 minutes, such as about 45 minutes.

[0118] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 45" includes ±5% of 45, or from 42.75 to 47.25.

[0119] In a preferred embodiment, the decocting vessel is covered when boiling.

[0120] In a preferred embodiment, the power of the container during boiling is 1000-3000W, for example, about 2000W.

[0121] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 2000" includes ±5% of 2000, or from 1900 to 2100.

[0122] In a preferred embodiment, the boiling time is 5 to 15 minutes, such as about 10 minutes.

[0123] 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.

[0124] In a preferred embodiment, the power of the container when slightly boiling is 500-1500W, such as about 900W.

[0125] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 900" includes ±5% of 900, or from 855 to 945.

[0126] In a preferred embodiment, the slight boiling time is 10 to 100 minutes, for example, about 60 minutes.

[0127] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 60" includes ±5% of 60, or from 57 to 63.

[0128] In a preferred embodiment, the volume / volume ratio of the decoction of the traditional Chinese medicine composition comprising angelica and cinnamon to the water is 0.1-0.5, such as about 0.3.

[0129] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.3" includes ±5% of 0.3, or from 0.285 to 0.315.

[0130] In a preferred embodiment, the filter is a 200 mesh nylon filter.

[0131] In a preferred embodiment, the concentration of ferulic acid in the reference solution is 10-25 μg / ml, such as about 18 μg / ml.

[0132] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 18" includes ±5% of 18, or from 17.1 to 18.9.

[0133] In a preferred embodiment, the concentration of cinnamaldehyde in the reference solution is 10-20 μg / ml, such as about 13 μg / ml.

[0134] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 13" includes ±5% of 13, or from 12.35 to 13.65.

[0135] In a preferred embodiment, the concentration of 6-gingerol in the reference solution is 1-10 μg / ml, such as about 6 μg / ml.

[0136] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 6" includes ±5% of 6, or from 5.7 to 6.3.

[0137] In a preferred embodiment, the concentration of ligustilide in the reference solution is 10-25 μg / ml, for example, about 18 μg / ml.

[0138] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 18" includes ±5% of 18, or from 17.1 to 18.9.

[0139] In a preferred embodiment, the gallic acid in the reference solution is 20-40 μg / ml, for example, about 30 μg / ml.

[0140] 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.

[0141] In a preferred embodiment, the concentration of paeoniflorin in the reference solution is 50-70 μg / ml, for example, about 60 μg / ml.

[0142] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 60" includes ±5% of 60, or from 57 to 63.

[0143] In a preferred embodiment, the cinnamic acid in the reference solution is 40-60 μg / ml, such as about 50 μg / ml.

[0144] 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.

[0145] In a preferred embodiment, the glycyrrhizic acid in the reference solution is 40-60 μg / ml, for example, about 50 μg / ml.

[0146] 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.

[0147] 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.

[0148] 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.

[0149] In a preferred embodiment, the column temperature is 20-30°C, such as about 25°C.

[0150] 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.

[0151] In a preferred embodiment, between the 0th minute and the 77th minute, the detection wavelength is 210-280 nm, such as about 250 nm.

[0152] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 250" includes ±5% of 250, or from 237.5 to 262.5.

[0153] In a preferred embodiment, between the 77th minute and the 100th minute, the detection wavelength is 190-230 nm, for example, about 210 nm.

[0154] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 210" includes ±5% of 210, or from 199.5 to 220.5.

[0155] In a preferred embodiment, the injection volume is 5-15 μl, such as about 10 μl.

[0156] 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.

[0157] In a preferred embodiment, the theoretical plate number of the chromatographic peak corresponding to the ferulic acid is not less than 5000.

[0158] In a preferred embodiment, the chromatographic column is an Agilent 5TC-C18 chromatographic column.

[0159] 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.

[0160] In a preferred embodiment, mobile phase A is acetonitrile.

[0161] In a preferred embodiment, the acid aqueous solution, base aqueous solution and / or buffered saline solution is selected from one or more of weak acids and their salts, weak bases and their salts of different concentrations.

[0162] In a preferred embodiment, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution is 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 in different concentrations.

[0163] In a preferred embodiment, the acid aqueous solution is a 0.01% to 1% acid aqueous solution.

[0164] In a preferred embodiment, the acid aqueous solution is a 0.01% to 1% phosphoric acid aqueous solution.

[0165] In a preferred embodiment, the aqueous acid solution is about 0.1% phosphoric acid aqueous solution.

[0166] 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%.

[0167] In a preferred embodiment, the buffered saline solution is an acetate aqueous solution and / or an acetate aqueous solution.

[0168] In a preferred embodiment, the pH of the buffered saline solution is no greater than 7.0.

[0169] In a preferred embodiment, when the detection wavelength between the 0th minute and the 77th minute is 250nm and the detection wavelength between the 77th minute and the 100th minute is 210nm, the fingerprint spectrum includes peaks 1-14, and the retention times of peaks 1-14 correspond to about 8.968min, about 21.263min, about 27.481min, about 29.945min, about 38.938min, about 42.241min, about 45.390min, about 64.056min, about 65.959min, about 69.988min, about 72.691min, about 74.593min, about 78.837min, and about 85.261min, respectively.

[0170] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 8.968" includes ±5% of 8.968, or from 8.5196 to 9.4164; "about 21.263" includes ±5% of 21.263, or from 20.19985 to 22.32615; "about 27.481" includes ±5% of 27.481, or from 26.10695 to 28.85505; "about 29.945" includes ±5% of 29.945 , or from 28.44775 to 31.44225; "about 38.938" includes ±5% of 38.938, or from 36.9911 to 40.8849; "about 42.241" includes ±5% of 42.241, or from 40.12895 to 44.35305; "about 45.390" includes ±5% of 45.390, or from 43.1205 to 47.6595; " "About 64.056" includes ±5% of 64.056, or from 60.8532 to 67.2588; "about 65.959" includes ±5% of 65.959, or from 62.66105 to 69.25695; "about 69.988" includes ±5% of 69.988, or from 66.4886 to 73.4874; "about 72.691" includes ±5% of 72.691, or From 69.05645 to 76.32555; "about 74.593" includes ±5% of 74.593, or from 70.86335 to 78.32265; "about 78.837" includes ±5% of 78.837, or from 74.89515 to 82.77885; "about 85.261" includes ±5% of 85.261, or from 80.99795 to 89.52405.

[0171] In a preferred embodiment, when the detection wavelength between the 0th minute and the 77th minute is 250 nm and the detection wavelength between the 77th minute and the 100th minute is 210 nm, the fingerprint spectrum includes peaks 1-14, wherein peak 4 is used as a reference peak, and the relative retention time averages of peak 1, peak 2, peak 3, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, and peak 14 are approximately 0.299, approximately 0.710, approximately 0.918, approximately 1.300, approximately 1.411, approximately 1.516, approximately 2.139, approximately 2.203, approximately 2.337, approximately 2.427, approximately 2.491, approximately 2.633, and approximately 2.847, respectively.

[0172] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.299" includes ±5% of 0.299, or from 0.28405 to 0.31395; "about 0.710" includes ±5% of 0.710, or from 0.6745 to 0.7455; "about 0.918" includes ±5% of 0.918, or from 0.8721 to 0.9639; "about 1.300" includes ±5% of 1.300, or from 1.235 to 1.365; "about 1.411" includes ±5% of 1.411, or from 1.34045 to 1.48155; "about 1.516" includes ±5% of 1.516, or from 1.4402 to 1.5918; "about 2.139" includes ±5% of 2.139, or from 2.03205 to 2.24595; “about 2.203” includes ±5% of 2.203, or from 2.09285 to 2.31315; “about 2.337” includes ±5% of 2.337, or from 2.22015 to 2.45385; “about 2.427” includes ±5% of 2.427, or from 2.30565 to 2.54835; “about 2.491” includes ±5% of 2.491, or from 2.36645 to 2.61555; “about 2.633” includes ±5% of 2.633, or from 2.50135 to 2.76465; “about 2.847” includes ±5% of 2.847, or from 2.70465 to 2.98935.

[0173] In a preferred embodiment, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, peak 1 is gallic acid, peak 4 is paeoniflorin, peak 5 is ferulic acid, peak 8 is cinnamic acid, peak 9 is cinnamaldehyde, peak 12 is glycyrrhizic acid, peak 13 is 6-gingerol, and peak 14 is ligustilide.

[0174] In a preferred embodiment, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, peak No. 1 comes from white peony medicinal material, peak No. 2 comes from white peony medicinal material, peak No. 3 comes from white peony medicinal material, peak No. 4 comes from white peony medicinal material, peak No. 5 comes from angelica medicinal material, peak No. 6 comes from stir-fried licorice medicinal material, peak No. 7 comes from white peony medicinal material, peak No. 8 comes from cinnamon medicinal material, peak No. 9 comes from cinnamon medicinal material, peak No. 10 comes from stir-fried licorice medicinal material, peak No. 11 comes from stir-fried licorice medicinal material, peak No. 12 comes from stir-fried licorice medicinal material, peak No. 13 comes from ginger medicinal material, and peak No. 14 comes from angelica medicinal material.

[0175] According to another aspect of the present invention, a quality control method for a Chinese medicine composition containing angelica and cinnamon is provided, the quality control method comprising the following steps:

[0176] (1) establishing a standard fingerprint of a reference sample of a Chinese medicine composition according to the above-mentioned fingerprint construction method;

[0177] (2) taking a sample solution of the Chinese medicine composition and testing it according to the chromatographic conditions in the fingerprint spectrum construction method to obtain a fingerprint spectrum of the Chinese medicine composition sample to be tested; and

[0178] (3) Compare the fingerprint of the sample of the Chinese medicine composition obtained in step (2) with the standard fingerprint of the reference sample of the Chinese medicine composition obtained in step (1). If the fingerprint meets the requirements, it is a qualified product; if the fingerprint does not meet the requirements, it is an unqualified product.

[0179] In a preferred embodiment, the compliance requirements include one or more of the following:

[0180] (1) The fingerprint of the Chinese medicine composition sample to be tested exhibits 14 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 composition reference sample;

[0181] (2) With peak No. 4 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine composition 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 composition reference sample; and

[0182] (3) According to the similarity evaluation system of Chinese medicine chromatographic fingerprint, the similarity between the fingerprint of the Chinese medicine composition sample to be tested and the standard fingerprint of the Chinese medicine composition reference sample shall be calculated, and the similarity shall not be less than 0.90.

[0183] According to another aspect of the present invention, there is provided a use of the above construction method or the above quality control method in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine composition containing angelica and cinnamon.

[0184] 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.

[0185] 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.

[0186] 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.

[0187] Example

[0188] Example

[0189] 1. Instruments and reagents

[0190] 1.1 Instruments

[0191] High performance liquid chromatograph, model: Agilent1290_DAD HPLC (Agilent Technologies Co., Ltd.); High performance liquid chromatograph, model: Waters 2695_2996 (Waters Technologies Co., Ltd.); Chromatographic column: Agilent 5TC-C18(2)250×4.6mm, SN:731424 / 735516 / 735519; Electronic balance, model: BCE224I-1CCN (Sartorius Group, Germany); Electronic balance model: JY2002 (Shanghai Puchun Metrology Instrument Co., Ltd.); Desktop low-speed centrifuge, model: TD4N (Changsha Yingtai Instrument Co., Ltd.); CNC ultrasonic cleaner, model: KQ-500DE (Kunshan Ultrasonic Instrument Co., Ltd.); Digital display constant temperature water bath, model: HH-6 (Shanghai Boxun Medical Biological Instrument Co., Ltd.); Diaphragm oil-free vacuum pump, model: MT-10 (Yancheng Xinmingte Glass Instrument Co., Ltd.).

[0192] 1.2 Reagents and test drugs

[0193] Methanol and acetonitrile were of chromatographic grade, methanol and phosphoric acid were of analytical grade, and water was purified water; Ferulic acid (batch number: 110773-202316, content: 99.3%, China Food and Drug Inspection Institute); Cinnamaldehyde (batch number: 110710-202223, content: 98.9%, China Food and Drug Inspection Institute); Ligustilide (batch number: 111737-202311, content: 100.0%, China Food and Drug Inspection Institute); 6-gingerol (batch number: 111833-202007, content: 99.3%) , China Food and Drug Inspection Institutes); gallic acid (batch number: 110831-201906, content: 91.5%, China Food and Drug Inspection Institutes); ammonium glycyrrhizinate (batch number: 110731-202122, content: 94.4%, China Food and Drug Inspection Institutes); cinnamic acid (batch number: 110786-202305, content: 99.8%, China Food and Drug Inspection Institutes); paeoniflorin (batch number: 110736-202447, content: 98.1%, China Food and Drug Inspection Institutes).

[0194] 1.3. Sample information

[0195] The Chinese medicine composition granule reference sample containing angelica and cinnamon of the present invention (batch number: 2024062801-L source: homemade); the Chinese medicine composition granule reference sample containing angelica and cinnamon of the present invention (batch number: 2024072501-L source: homemade); the Chinese medicine composition granule reference sample containing angelica and cinnamon of the present invention (batch number: 2024082904-L source: homemade); the Chinese medicine composition granule containing angelica and cinnamon of the present invention lacking angelica negative reference sample (batch number: 202409 0201-L Source: homemade); the Chinese medicine composition granules containing angelica and cinnamon of the present invention lack cinnamon negative benchmark sample (batch number: 2024090204-L Source: homemade); the Chinese medicine composition granules containing angelica and cinnamon of the present invention lack white peony negative benchmark sample (batch number: 2024090202-L Source: homemade); the Chinese medicine composition granules containing angelica and cinnamon of the present invention lack ginger negative benchmark sample (batch number: 2024090301-L Source: homemade); the Chinese medicine composition granules containing angelica and cinnamon of the present invention lack ginger negative benchmark sample (batch number: 2024090302-L Source: homemade); The Chinese medicine composition granules containing angelica and cinnamon lack the negative reference sample of jujube (batch number: 2024090302-L source: homemade); the Chinese medicine composition granules containing angelica and cinnamon of the present invention lack the negative reference sample of fried licorice (batch number: 2024090203-L source: homemade); angelica slices (batch number: ZA00323102101-01, origin: Min County, Dingxi City, Gansu Province, source: homemade); cinnamon slices (batch number: ZA04223112801, origin: Ping, Guigang City, Guangxi Zhuang Autonomous Region, source : homemade); white peony slices (batch number: ZA00123121502, origin: Qiaocheng District, Bozhou City, Anhui Province, source: homemade); jujube slices (batch number: ZA00223121402, origin: Ningyang County, Tai'an City, Shandong Province, source: homemade); ginger slices (batch number: ZA00524071501, origin: Wanyuan County, Dazhou City, Sichuan Province, source: homemade); stir-fried licorice slices (batch number: ZD00423090401-01, origin: Hangjin Banner, Ordos, Inner Mongolia, source: homemade).

[0196] 1.4. Solution preparation

[0197] (1) Preparation method of reference sample decoction

[0198] Take 55.20g of angelica, 41.40g of cinnamon, 82.80g of white peony root, 41.40g of ginger, 36.00g of jujube, and 27.60g of stir-fried licorice, grind them into coarse particles, put them in a ceramic decoction pot, add 2000ml of water, cover, soak for 45 minutes, boil at 2000W in an electric ceramic stove for about 10 minutes, keep it at a slight boil at 900W and simmer until 600ml (about 60 minutes), filter through a 200-mesh nylon filter while hot.

[0199] (2) Preparation of reference solution:

[0200] Take appropriate amounts of ferulic acid, cinnamaldehyde, 6-gingerol, ligustilide, gallic acid, ammonium glycyrrhizinate, cinnamic acid, and paeoniflorin reference substances, weigh them accurately, and add methanol to make a mixed solution containing 18 μg of ferulic acid, 13 μg of cinnamaldehyde, 6 μg of 6-gingerol, 18 μg of ligustilide, 30 μg of gallic acid, 50 μg of ammonium glycyrrhizinate, 50 μg of cinnamic acid, and 60 μg of paeoniflorin per 1 ml as the reference solution.

[0201] (3) Preparation of test solution

[0202] Take about 15g of the reference sample decoction, place it in a 25ml volumetric flask, add an appropriate amount of methanol, ultrasonically (500W, 40kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000rpm / min for 10min, filter the supernatant, and take the filtrate to obtain the product.

[0203] (4) Preparation of negative control solution

[0204] Preparation of negative sample solution lacking Angelica sinensis: weigh other medicinal ingredients lacking Angelica sinensis according to the prescription, prepare negative sample lacking Angelica sinensis according to the reference sample preparation method, and prepare negative sample solution lacking Angelica sinensis according to item "(3)".

[0205] Negative control solutions lacking angelica, cinnamon bark, white peony root, ginger, jujube, and fried licorice were prepared in the same way.

[0206] (5) Blank solvent: methanol.

[0207] 3. Establishment of feature map method

[0208] 3.1 Study on characteristic spectrum chromatographic conditions

[0209] The proposed initial chromatographic conditions are:

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

[0211] Chromatographic conditions and system suitability test: Octadecyl bonded silica gel was used as filler (Agilent 5TC-C18 4.6×250 mm 5 μm); acetonitrile was used as mobile phase A, 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in Table 1; the flow rate was 1 ml per minute; the column temperature was 25° C.; and full wavelength scanning was performed.

[0212] Table 1 Characteristic spectrum of the Chinese medicine composition particles containing angelica and cinnamon of the present invention Chromatographic condition gradient 1

[0213]

[0214]

[0215] Preparation of reference solution: Take appropriate amount of ferulic acid, paeoniflorin, cinnamaldehyde, 6-gingerol, ligustilide, gallic acid, ammonium glycyrrhizinate, cinnamic acid, and paeoniflorin reference substances, weigh them accurately, and add methanol to prepare a mixed solution containing 18 μg of ferulic acid, 1.0 mg of paeoniflorin, 13 μg of cinnamaldehyde, 6 μg of 6-gingerol, 18 μg of ligustilide, 30 μg of gallic acid, 50 μg of ammonium glycyrrhizinate, 50 μg of cinnamic acid, and 60 μg of paeoniflorin per 1 ml as the reference solution.

[0216] Preparation of test solution: Take about 15 g of the decoction, place it in a 25 ml volumetric flask, add an appropriate amount of methanol, ultrasonically (500 W, 40 kHz) for 30 minutes, cool, make up to volume, shake well, filter, and take the filtrate to obtain the test solution.

[0217] Determination method: Accurately pipette 10μl of reference solution and test solution, inject into liquid chromatograph, and determine.

[0218] The summary table of the optimal absorption wavelengths of the reference solution is shown in Table 2.

[0219] Table 2 Summary of the best absorption wavelengths of reference solutions

[0220] Reference solution Peak time Optimum absorption wavelength (nm) gallic acid 7.531 216、272 Paeoniflorin 28.460 232 Ferulic acid 34.661 218、236、322 Chuanxiong lactone I 50.274 278 Cinnamaldehyde 61.018 206、222、290 Glycyrrhizic acid 73.644 252 6-Gingerol 77.183 282 Ligustilide 84.164 208、282、326

[0221] The results show that, combined with the full-wavelength UV absorption spectrum, there is UV absorption fluorescence at a wavelength of 200nm to 300nm; compared with the liquid phase spectra at 210nm, 250nm and 254nm, the response values ​​of the characteristic peaks at 254nm and 250nm wavelengths are significantly better than those at 210nm within 0 to 75 minutes; 6-gingerol (t R =77.321) and ligustilide (t R =84.083); therefore, the characteristic spectrum detection wavelength of the Chinese medicine composition granule benchmark sample containing angelica and cinnamon of the present invention is determined as follows: the detection wavelength within 0 to 77 minutes is 250nm, and the detection wavelength within 77 to 100 minutes is 210nm. In view of the poor separation of the characteristic peaks at 30 to 40 minutes, it is further optimized to the gradient 2 of Table 3.

[0222] Table 3 Gradient 2

[0223] Time (min) Acetonitrile (%) 0.1% Phosphoric acid (%) 0→10 5→6 95→94 10→12 6→12 94→88 12→20 12 88 20→21 12→16 88→84 21→38 16 84 38→48 16→20 84→80 48→52 20→25 80→75 52→58 25→29 75→71 58→62 29→32 71→68 62→80 32→56 68→44 80→83 56→74 44→26 83→85 74→5 26→95 85→100 5 95

[0224] The results show that the separation and response values ​​of each characteristic peak under gradient 2 have reached the requirements of the liquid phase characteristic spectrum, so this gradient is determined as the characteristic spectrum method of the Chinese medicine composition granule reference sample containing Angelica and cinnamon of the present invention; the detection wavelength is: the detection wavelength within 0-77 minutes is 250nm, and the detection wavelength within 77-100 minutes is 210nm. The chromatogram of the Chinese medicine composition granule decoction reference sample containing Angelica and cinnamon of the present invention presents 21 chromatographic peaks. In view of the low peak area response values ​​of peaks 7, 9-12, and 14, and the poor peak shape of peak 18, considering the batch difference, it is not included in the characteristic spectrum characteristic peak standard; the response values ​​and separation of the remaining chromatographic peaks are good, so it is determined that the characteristic spectrum of the Chinese medicine composition granule decoction reference sample containing Angelica and cinnamon of the present invention includes 14 characteristic peaks. The peak time of peak 21 (ligustilide) is about 85 minutes and has returned to the initial gradient, and is further optimized to gradient 3 in Table 4.

[0225] Table 4 Gradient 3

[0226]

[0227]

[0228] The results show that the separation and response values ​​of each characteristic peak under this gradient have reached the requirements of the liquid phase characteristic spectrum, so this gradient is determined as the characteristic spectrum method of the reference sample of the Chinese medicine composition granules containing angelica and cinnamon of the present invention; the detection wavelength is: the detection wavelength within 0 to 77 minutes is 250nm, and the detection wavelength within 77 to 100 minutes is 210nm.

[0229] 3.2. Preparation of reference sample test solution

[0230] According to the results of the research on the preparation process of the reference sample, the reference sample was stored in the form of a decoction, so the preparation method of the decoction test solution was investigated. Therefore, it was determined that 15g of the decoction was taken for the research.

[0231] 3.2.1. Investigation of extraction solvent

[0232] Six portions of the decoction were taken to investigate the effects of pure water, 50% methanol and methanol as extraction solvents on the characteristic spectra of the reference samples of the Chinese medicine composition granules containing angelica and cinnamon bark of the present invention.

[0233] Take 6 portions of the reference sample decoction, about 15 g each, weigh accurately, put into a 25 ml volumetric flask, add pure water, 50% methanol, and appropriate amount of methanol respectively, treat with ultrasound (500W, 40kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000rpm / min for 10 minutes, take the supernatant for filtration, and take the filtrate to obtain the product.

[0234] The results show that the number of characteristic peaks of the Chinese medicine composition particles containing angelica and cinnamon bark treated with different extraction solvents is the same, and the total peak area is slightly larger after methanol treatment, so the treatment solvent is methanol.

[0235] 3.2.2. Extraction time investigation

[0236] Six portions of the decoction were taken to examine the effects of extraction times of 20 minutes, 30 minutes, and 45 minutes on the characteristic spectra of the reference samples of the Chinese medicine composition granules containing angelica and cinnamon of the present invention.

[0237] Take 6 portions of the reference sample decoction, about 15 g each, weigh accurately, place in a 25 ml volumetric flask, add an appropriate amount of methanol, treat with ultrasound (500 W, 40 kHz) for 20 min, 30 min, and 45 min respectively, cool, make up to volume, shake well, centrifuge at 5000 rpm / min for 10 min, filter the supernatant, and take the filtrate to obtain the product.

[0238] The results show that after the Chinese medicine composition particles containing Angelica sinensis and cinnamon bark are processed for different extraction times, the number of characteristic peaks is the same and the total peak area is close. Considering the continuity of the experiment, the extraction time is 30 minutes.

[0239] 3.2.3. Sampling volume investigation

[0240] Six portions of the decoction were taken to investigate the effects of sampling amounts of 10 g, 15 g, and 20 g on the characteristic spectrum of the benchmark sample of the Chinese medicine composition granules comprising angelica and cinnamon bark of the present invention.

[0241] Take 6 portions of the reference sample decoction, about 10g, 15g, and 20g each, weigh accurately, place in a 25ml volumetric flask, add appropriate amount of methanol, treat with ultrasound (500W, 40kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000rpm / min for 10min, filter the supernatant, and take the filtrate to obtain the product.

[0242] The results show that after different sampling amounts of the Chinese medicine composition particles containing angelica and cinnamon bark are treated with methanol, the total peak area of ​​the sampling amount is 10-20g and changes proportionally, indicating that the detection method is stable within the range of 10-20g. Considering that the content detection and the characteristic spectrum detection share the same method, the sampling amount is determined to be 15g.

[0243] In summary, the method for preparing the test solution of the characteristic spectrum of the benchmark sample of the Chinese medicine composition granules containing angelica and cinnamon of the present invention is: take about 15g of the benchmark sample decoction, accurately weigh it, put it in a 25ml volumetric flask, add an appropriate amount of methanol, ultrasonically (500W, 40kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000rpm / min for 10min, filter the supernatant, and take the filtrate to obtain.

[0244] 4. Methodological Validation

[0245] 4.1. Exclusivity

[0246] The blank solvent, the reference solution, the negative control solution of each medicinal flavor of the benchmark sample solution of the Chinese medicine composition granules containing Angelica sinensis and cinnamon bark of the present invention, and the single medicinal material control solution are sampled and analyzed. The blank solvent must not interfere with the proposed characteristic peak position. In addition, the attribution of the chromatographic peak must be determined based on the test results of the negative control solution and the single medicinal material control solution. Figure 1 It is the liquid phase fingerprint of the reference sample of the Chinese medicine composition granules containing angelica and cinnamon of the present invention.

[0247] In summary, from the results of the attribution identification of the chromatographic peaks of the characteristic spectrum of the granules of the Chinese medicine composition containing angelica and cinnamon of the present invention, it can be seen that the chromatogram of the reference sample of the granule decoction of the Chinese medicine composition containing angelica and cinnamon of the present invention mainly presents 14 chromatographic peaks, among which peaks 1 to 4 and peak 7 belong to the medicinal taste of white peony, among which peak 1 is gallic acid and peak 4 is paeoniflorin; peaks 5 and 14 belong to the medicinal taste of angelica, which are ferulic acid and ligustilide, respectively; peaks 6 and 10 to 12 belong to the medicinal taste of stir-fried licorice, among which peak 12 is glycyrrhizic acid; peaks 8 and 9 belong to the medicinal taste of cinnamon, among which peak 8 is cinnamic acid and peak 9 is cinnamaldehyde; peak 13 belongs to the medicinal taste of ginger, which is 6-gingerol.

[0248] The blank solvent, the reference solution, the negative control solution of each medicinal flavor of the Chinese medicine composition granule reference sample solution containing angelica and cinnamon of the present invention, and the single taste control solution were sampled and analyzed. At the proposed characteristic peak position, the blank solvent and the negative control solution had no interference. The specific medicinal flavors of the chromatographic peaks in the chromatogram are shown in Table 5.

[0249] Table 5 Details of the medicinal flavors attributed to specific chromatographic peaks in the chromatogram

[0250] Medicinal taste Angelica Cinnamon White Peony Root Ginger Jujube Fried Licorice Remark Attribution Peak 1 - - + - - - gallic acid White Peony Root Peak 2 - - + - - - / White Peony Root Peak 3 - - + - - - / White Peony Root Peak 4 - - + - - - Paeoniflorin White Peony Root Peak 5 + - - - - - Ferulic acid Angelica Peak 6 - - - - - + / Fried Licorice Peak 7 - - + - - - / White Peony Root Peak 8 - + - - - - Cinnamic acid Cinnamon Peak 9 - + - - - - Cinnamaldehyde Cinnamon Peak 10 - - - - - + / Fried Licorice Peak 11 - - - - - + / Fried Licorice Peak 12 - - - - - + Glycyrrhizic acid Fried Licorice Peak 13 - - - + - - 6-Gingerol Ginger Peak 14 + - - - - - Ligustilide Angelica

[0251] 4.2 Precision

[0252] 4.2.1 Repeatability

[0253] According to the above test solution preparation method, 6 test solutions were prepared, sampled and tested, chromatograms were recorded, and the RSD value of the relative retention time between the characteristic peak and the S peak (peony root glucoside) was calculated; the RSD of the relative retention time was ≤3%.

[0254] The results showed that the RSD of the relative retention time of each characteristic peak in the repeatability experiment was ≤3%, indicating that this method had good repeatability.

[0255] 4.2.2 Intermediate precision

[0256] Two personnel prepared 6 test solutions according to the above conditions, respectively, and injected and tested them on Agilent HPLC and Waters HPLC, and calculated the RSD value of the relative retention time of each characteristic peak and the S peak (peony indole); if the RSD of the relative retention time is ≤3%, it means that the intermediate precision is good.

[0257] The results showed that the RSD of the relative retention time of each characteristic peak between Waters and Agilent was ≤3%, indicating that this method had good precision between Waters and Agilent.

[0258] 4.2.3 Instrument Precision

[0259] Prepare a sample solution according to the above conditions, inject 6 times continuously, record the chromatogram, calculate the RSD value of the retention time and peak area of ​​the characteristic peak and S peak (peony glucoside); the RSD of the relative retention time is ≤3%, the RSD of the relative peak area is ≤5%, and the relative peak area of ​​the chromatographic peak with a small proportion of the total peak area can be appropriately relaxed. This indicates that the instrument has good precision.

[0260] The results showed that the RSD of the relative retention time of each characteristic peak in the instrument precision experiment was ≤3%; the RSD of the relative peak area of ​​each characteristic peak was ≤5%, indicating that the instrument precision was good.

[0261] 4.3 Durability

[0262] 4.3.1 Flow rate durability

[0263] Prepare one sample solution according to the above sample solution preparation method, set the mobile phase flow rate to 0.9 ml / min, 1.0 ml / min, and 1.1 ml / min respectively, inject and detect, record the chromatogram, calculate the relative average deviation of the relative retention time of each characteristic peak, and the relative average deviation of the relative retention time is ≤3%, which indicates that the flow rate durability is good.

[0264] The results show that the relative average deviation of the relative retention time of each characteristic peak in different flow rate experiments is ≤3%, except for peak 1. In view of the long running time of this gradient, it can be appropriately relaxed, indicating that the flow rate durability is good.

[0265] 4.3.2 Column temperature durability

[0266] Prepare 1 sample solution according to the above sample solution preparation method, inject and detect at column temperatures of 20℃, 25℃, and 30℃, record the chromatogram, and calculate the relative average deviation of the relative retention time of each characteristic peak. If the relative average deviation of the relative retention time is ≤3%, it means that the flow rate durability is good.

[0267] The results show that the relative average deviation of the relative retention time of each characteristic peak in different column temperature experiments is ≤3%, except for peak 1. In view of the long running time of this gradient, it can be appropriately relaxed, indicating that the column temperature has good durability.

[0268] 4.3.3 Durability at different acid concentrations

[0269] Prepare one sample solution according to the above sample solution preparation method, inject and detect it in 0.09%, 0.10% and 0.11% phosphoric acid solution, record the chromatogram, calculate the relative average deviation of the relative retention time of each characteristic peak, and if the relative average deviation of the relative retention time is ≤3%, it means that the durability of different acid concentrations is good.

[0270] The results show that the relative average deviation of the relative retention time of each characteristic peak in experiments with different acid concentrations is ≤3%, indicating that the durability of different acid concentrations is good.

[0271] 4.3.4 Durability of different chromatographic columns

[0272] Prepare one sample solution according to the above sample solution preparation method, use three numbered chromatographic columns respectively, record the chromatograms, calculate the relative average deviation of the relative retention time of each characteristic peak, and the relative average deviation of the relative retention time is ≤3%, which means that the durability of different chromatographic columns is good.

[0273] The results showed that the relative average deviation of the relative retention time of each characteristic peak in different chromatographic column experiments was ≤3%, indicating that different chromatographic columns had good durability.

[0274] 4.4. Stability of test solution

[0275] Prepare 1 sample solution according to the above sample solution preparation method, and inject and test at 0, 3, 6, 10, 15, 18, 24, 36, 48, and 62 hours respectively. Record the chromatogram, calculate the relative retention time and RSD value of the relative peak area of ​​each characteristic peak; the relative peak area of ​​the chromatographic peak with a small proportion of the total peak area can be appropriately relaxed. This indicates that the stability of the sample solution is good.

[0276] The results show that the RSD of the relative retention time of each characteristic peak in the stability test of the test solution at 0, 3, 6, 10, 15, 18, 24, 36, 48, and 62 hours is ≤3%; the RSD of the relative peak area is ≤5%, except for peak 9, which can be relaxed in view of the fact that the total peak area of ​​peak 9 accounts for ≤1.0%. This shows that the test solution has good stability within 62 hours.

[0277] 5. Confirmation of the characteristic spectrum method of the benchmark sample

[0278] Based on the above research results, according to the guidelines for validation of analytical methods in General Chapter 9101 of the 2020 edition of the "Chinese Pharmacopoeia", the specificity, accuracy, precision, column temperature, flow rate, different acid concentrations, different chromatographic columns, and stability of the test solution of this characteristic detection method all meet the requirements, so this detection method is better, as follows.

[0279] Determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Rules 0512).

[0280] Chromatographic conditions and system suitability test: Octadecyl bonded silica gel was used as filler (Agilent 5TC-C18 4.6×250mm 5μm); acetonitrile was used as mobile phase A, 0.1% phosphoric acid solution was used as mobile phase B, and gradient elution was performed according to the provisions in Table 4; the flow rate was 1 ml per minute; the column temperature was 25°C; the detection wavelength within 0 to 77 minutes was 250 nm, and the detection wavelength within 77 to 100 minutes was 210 nm. The theoretical plate number calculated based on the ferulic acid peak should not be less than 5000.

[0281] Preparation of reference solution: Take appropriate amounts of ferulic acid, cinnamaldehyde, 6-gingerol, ligustilide, gallic acid, ammonium glycyrrhizinate, cinnamic acid, and paeoniflorin reference substances, weigh them accurately, and add methanol to prepare a mixed solution containing 18 μg of ferulic acid, 13 μg of cinnamaldehyde, 6 μg of 6-gingerol, 18 μg of ligustilide, 30 μg of gallic acid, 50 μg of ammonium glycyrrhizinate, 50 μg of cinnamic acid, and 60 μg of paeoniflorin per 1 ml as the reference solution.

[0282] Preparation of test solution: Take about 15 g of the reference sample decoction, weigh it accurately, put it in a 25 ml volumetric flask, add an appropriate amount of methanol, sonicate (500 W, 40 kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000 rpm / min for 10 minutes, filter the supernatant, and take the filtrate to obtain the test solution.

[0283] Determination method: Accurately pipette 10μl of reference solution and test solution, inject into liquid chromatograph, and determine.

[0284] 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 medicine composition containing angelica and cinnamon, characterized in that: The construction method comprises the following steps: Preparation of a Chinese medicine composition test solution: taking an appropriate amount of a decoction of a Chinese medicine composition containing angelica and cinnamon, placing it in a container, adding a solvent, ultrasonically treating it for a period of time, fixing the volume, shaking it well, centrifuging it, filtering the supernatant, and taking the filtrate to obtain the Chinese medicine composition test solution; wherein the Chinese medicine composition includes angelica, cinnamon, white peony root, ginger, jujube and liquorice; Preparation of reference solution: weigh appropriate amounts of gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide reference substances, add methanol to prepare reference solutions with concentrations of 1 to 100 μg / ml for gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide, respectively; Obtaining a fingerprint of the Chinese medicine composition according to the results of high performance liquid chromatography detection of the Chinese medicine composition test solution and the reference solution; The chromatographic conditions of the high performance liquid phase detection are as follows: a chromatographic column with octadecyl bonded silica gel as filler is used, mobile phase A is selected from one or more of acetonitrile, methanol and tetrahydrofuran, mobile phase B is an acid aqueous solution, an alkaline aqueous solution and / or a buffered saline solution, and the gradient elution program is: 0-10 min, 95% → 94% B; 10-12 min, 94% B → 88% B; 12-20 min, 88% B; 20-21 min, 88% → 84% B; 21-38 min, 84% B; 38-48 min, 84% B → 80% B; 48-5 2min, 80%→75%B; 52~58min, 75%B→71%B; 58~62min, 71%→68%B; 62~80min, 68%B→44%B; 80~83min, 44%B→26%B; 83~88min, 26%B→15%B; 88~89min, 15%B→95%B; 89~100min, 95%B; the flow rate is 0.5~1.5ml / min, the column temperature is 10~40℃, the detection wavelength is 180~350nm, and the injection volume is 1~20μl.

2. The construction method according to claim 1, characterized in that: The licorice is stir-fried licorice.

3. The construction method according to claim 1, characterized in that: In the preparation of the test solution of the Chinese medicine composition, the solvent is an alcohol, such as methanol; Preferably, the container is a volumetric flask; Preferably, the mass of the decoction of the Chinese medicine composition is 5 to 30 g, for example, about 15 g; Preferably, the mass volume ratio between the decoction of the traditional Chinese medicine composition and the solvent is 0.4 to 0.8, such as about 0.6, in units of g / ml; More preferably, the power of the ultrasonic treatment is 400-600W, for example about 500W; More preferably, the frequency of the ultrasonic treatment is 30 to 50 kHz, such as about 40 kHz; More preferably, the ultrasonic treatment time is 20 to 40 min, for example about 30 min; Preferably, the centrifugal speed is 4000-6000 rpm / min, for example, about 5000 rpm / min; Preferably, the centrifugation time is 5 to 15 minutes, for example, about 10 minutes; Particularly preferably, the mass ratio of the angelica sinensis, the cinnamon bark, the white peony root, the ginger, the jujube and the liquorice is (45-60):(30-50):(70-90):(30-50):(25-45):(20-40); Particularly preferably, the mass ratio of the angelica sinensis, the cinnamon bark, the white peony root, the ginger, the jujube and the liquorice is about 55.2: about 41.4: about 82.8: about 41.4: about 36: about 27.6; Particularly preferably, the preparation method of the decoction of the Chinese medicine composition containing angelica and cinnamon comprises: weighing appropriate amounts of angelica, cinnamon, white peony root, ginger, jujube and liquorice, crushing them into coarse particles, placing them in a container, soaking them in water, boiling them until boiling for a period of time, then keeping them slightly boiling for a period of time, filtering the filtrate with a filter while it is hot to obtain the decoction of the Chinese medicine composition containing angelica and cinnamon; Particularly preferably, the volume / mass ratio of the water to the Chinese medicinal composition comprising angelica and cinnamon is 5 to 10, for example, about 7; Particularly preferably, the container is a ceramic medicine decoction pot or an electric ceramic stove; Particularly preferably, the soaking time is 10 to 100 minutes, for example, about 45 minutes; Particularly preferably, the decocting is covered when boiling; Particularly preferably, the power of the container during boiling is 1000-3000W, for example about 2000W; Particularly preferably, the boiling time is 5 to 15 minutes, for example about 10 minutes; Particularly preferably, the power of the container at slight boiling is 500 to 1500 W, for example about 900 W; Particularly preferably, the slight boiling time is 10 to 100 minutes, for example about 60 minutes; Particularly preferably, the volume / volume ratio of the decoction of the Chinese medicinal composition comprising angelica and cinnamon to the water is 0.1 to 0.5, for example, about 0.3; Particularly preferably, the filter is a 200-mesh nylon filter.

4. The construction method according to claim 1, characterized in that: The concentration of ferulic acid in the reference solution is 10-25 μg / ml, for example, about 18 μg / ml; Preferably, the concentration of cinnamaldehyde in the reference solution is 10-20 μg / ml, for example, about 13 μg / ml; Preferably, the concentration of 6-gingerol in the reference solution is 1 to 10 μg / ml, for example, about 6 μg / ml; Preferably, the concentration of ligustilide in the reference solution is 10 to 25 μg / ml, for example, about 18 μg / ml; Preferably, the gallic acid in the reference solution is 20-40 μg / ml, for example, about 30 μg / ml; Preferably, the concentration of paeoniflorin in the reference solution is 50-70 μg / ml, for example, about 60 μg / ml; Preferably, the cinnamic acid in the reference solution is 40-60 μg / ml, for example, about 50 μg / ml; Preferably, the glycyrrhizic acid content in the reference solution is 40-60 μg / ml, for example, about 50 μg / ml.

5. The construction method according to claim 1, characterized in that: The flow rate of the HPLC detection is 0.8 to 1.2 ml / min, for example, about 1.0 ml / min; Preferably, the column temperature is 20-30°C, such as about 25°C; Preferably, between minute 0 and minute 77, the detection wavelength is 210 to 280 nm, for example, about 250 nm; Preferably, between the 77th minute and the 100th minute, the detection wavelength is 190 to 230 nm, for example, about 210 nm; Preferably, the injection volume is 5 to 15 μl, for example, about 10 μl; More preferably, the theoretical plate number of the chromatographic peak corresponding to the ferulic acid is not less than 5000; Preferably, the chromatographic column is an Agilent 5TC-C18 chromatographic column; Even more preferably, the specifications of the chromatographic column are: column length 250 mm, inner diameter 4.6 mm, and particle size 5 μm.

6. The construction method according to claim 1, characterized in that: Mobile phase A was acetonitrile; Preferably, the acid aqueous solution, base aqueous solution and / or buffered saline solution is selected from one or more of weak acids and their salts, weak bases and their salts of different concentrations; Preferably, the acid aqueous solution, alkali aqueous solution and / or buffered saline solution is 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 in different concentrations; More preferably, the acid aqueous solution is a 0.01% to 1% acid aqueous solution; More preferably, the acid aqueous solution is a 0.01% to 1% phosphoric acid aqueous solution; More preferably, the aqueous acid solution is an aqueous solution of about 0.1% phosphoric acid; Also preferably, the buffered saline solution is an acetate solution and / or an acetate solution; Also preferably, the pH value of the buffered saline solution is not greater than 7.

0.

7. The construction method according to claim 1, characterized in that: When the detection wavelength between the 0th minute and the 77th minute is 250nm and the detection wavelength between the 77th minute and the 100th minute is 210nm, the fingerprint spectrum includes peaks 1-14, and the retention times of the peaks 1-14 correspond to about 8.968min, about 21.263min, about 27.481min, about 29.945min, about 38.938min, about 42.241min, about 45.390min, about 64.056min, about 65.959min, about 69.988min, about 72.691min, about 74.593min, about 78.837min, and about 85.261min, respectively; Preferably, when the detection wavelength between the 0th minute and the 77th minute is 250 nm, and the detection wavelength between the 77th minute and the 100th minute is 210 nm, the fingerprint spectrum includes peaks 1-14, wherein peak 4 is used as a reference peak, and the relative retention time averages of peaks 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14 are about 0.299, about 0.710, about 0.918, about 1.300, about 1.411, about 1.516, about 2.139, about 2.203, about 2.337, about 2.427, about 2.491, about 2.633, and about 2.847, respectively; More preferably, when the detection wavelength between the 0th minute and the 77th minute is 250 nm, and the detection wavelength between the 77th minute and the 100th minute is 210 nm, peak 1 is gallic acid, peak 4 is paeoniflorin, peak 5 is ferulic acid, peak 8 is cinnamic acid, peak 9 is cinnamaldehyde, peak 12 is glycyrrhizic acid, peak 13 is 6-gingerol, and peak 14 is ligustilide; More preferably, when the detection wavelength between the 0th minute and the 77th minute is 250nm, and the detection wavelength between the 77th minute and the 100th minute is 210nm, peak 1 comes from white peony medicinal material, peak 2 comes from white peony medicinal material, peak 3 comes from white peony medicinal material, peak 4 comes from white peony medicinal material, peak 5 comes from angelica medicinal material, peak 6 comes from stir-fried licorice medicinal material, peak 7 comes from white peony medicinal material, peak 8 comes from cinnamon medicinal material, peak 9 comes from cinnamon medicinal material, peak 10 comes from stir-fried licorice medicinal material, peak 11 comes from stir-fried licorice medicinal material, peak 12 comes from stir-fried licorice medicinal material, peak 13 comes from ginger medicinal material, and peak 14 comes from angelica medicinal material.

8. A method for quality control of a Chinese medicine composition comprising angelica and cinnamon, characterized in that: The quality control method comprises the following steps: (1) Establishing a standard fingerprint of a reference sample of a Chinese medicine composition according to the fingerprint construction method described in any one of claims 1 to 7; (2) taking a sample solution of the Chinese medicine composition, and testing it according to the chromatographic conditions in the fingerprint construction method according to any one of claims 1 to 7 to obtain a fingerprint of the Chinese medicine composition sample to be tested; as well as (3) Compare the fingerprint of the Chinese medicine composition sample obtained in step (2) with the standard fingerprint of the Chinese medicine composition 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.

9. The quality control method according to claim 8, characterized in that: The compliance requirements include one or more of the following: (1) The fingerprint of the Chinese medicine composition sample to be tested exhibits 14 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 composition reference sample; (2) With peak No. 4 as the S peak, the relative retention time of each characteristic chromatographic peak in the fingerprint of the Chinese medicine composition 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 composition 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 composition sample to be tested and the standard fingerprint of the Chinese medicine composition reference sample is calculated, and the similarity shall not be less than 0.

90.

10. Use of the construction method according to any one of claims 1 to 7 or the quality control method according to claim 8 or 9 in quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine composition containing angelica and cinnamon.

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