A method for constructing a fingerprint of a traditional Chinese medicine composition containing angelica and cinnamon and application thereof

Abstract

This invention discloses a method for constructing a fingerprint spectrum of a traditional Chinese medicine composition containing Angelica sinensis and cinnamon. The method includes the following steps: preparation of a test solution; preparation of a reference solution; and obtaining a fingerprint spectrum of the traditional Chinese medicine composition based on high-performance liquid chromatography (HPLC) detection of the test solution and the reference solution. This invention provides a comprehensive and systematic analysis of the chemical components of a traditional Chinese medicine composition containing Angelica sinensis and cinnamon, offering a theoretical basis for in-depth research on quality control and the pharmacodynamic material basis.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a method for constructing fingerprint spectra of a traditional Chinese medicine composition containing angelica and cinnamon, and its application. Background Technology

[0002] According to the "Law of the People's Republic of China on Traditional Chinese Medicine," ancient classic prescriptions refer to "prescriptions recorded in ancient Chinese medicine classics that are still widely used today, have definite curative effects, and possess distinct characteristics and advantages." Ancient classic prescriptions have a long and rich history of use in my country and are still applied today. They are a summary of the essence of clinical practice by physicians throughout history, carrying the profound accumulation of thousands of years of brilliant Chinese medicine civilization. They represent the refinement of Chinese medicine theory over thousands of years, a summary of clinical experience throughout the ages, and the most essential part of the great treasure trove of Chinese medicine. In-depth research and development of classic Chinese medicine prescriptions is a golden key to unlocking the treasure trove of traditional Chinese medicine.

[0003] Traditional Chinese medicine decoctions, based on classic formulas, are the most commonly used dosage form in traditional Chinese medicine clinical practice. They possess advantages such as rational formulation, rapid onset of action, significant efficacy, and easy absorption, earning them widespread trust among patients. However, their drawbacks include susceptibility to spoilage and mold due to difficulties in preparation, carrying, temporary decoction, and prolonged storage; bitter taste and large quantities; and difficulty in standardizing procedures, all of which seriously affect their clinical efficacy and make them unsuitable for modern lifestyles. To maintain the advantages of decoctions and overcome their shortcomings, the Center for Drug Evaluation of the National Medical Products Administration issued the "Technical Guidelines for Pharmaceutical Research of Traditional Chinese Medicine Compound Preparations Managed According to the Catalogue of Ancient Classic Formulas (Trial Implementation)" on August 31, 2021. This guideline clearly states that the quality of traditional Chinese medicine compound preparations managed according to the catalogue of ancient classic formulas should be basically consistent with the quality of the reference sample of the classic formula. The reference sample represents the overall intrinsic quality of the preparation; except for the molding process, the reference sample and the preparation's other quality control indicators are basically consistent. Therefore, the reference sample is a physical comparison of the preparation's intrinsic quality and a reference for optimizing large-scale production processes and establishing quality standards. Reference samples serve as the benchmark for classic formulas and indeed for all traditional Chinese medicine research and development, acting as control substances to ensure the safety and efficacy of drugs. The formulation of production process routes, parameter optimization, and quality standard setting for compound preparations based on classic formulas must be based on these reference samples. Reference samples act as a link between clinical practice, industry, and scientific research, serving as a standard for the inheritance, application, and promotion of traditional Chinese medicine.

[0004] Fingerprinting, based on the understanding of the holistic effects of the chemical components of traditional Chinese medicine (TCM), utilizes spectroscopic and chromatographic techniques to obtain the spectra or chromatograms of TCM chemical components. It is a feasible model for identifying the authenticity of TCM, evaluating quality consistency, and assessing product stability, characterized by its large information content, strong characteristic features, holistic nature, and fuzziness. TCM fingerprinting can comprehensively reflect the relative relationships of the chemical components contained in medicinal materials, demonstrating the complexity and correlation of TCM components. It aligns with traditional TCM theories and can effectively characterize, comprehensively evaluate, and fully control the intrinsic quality of TCM. It is particularly suitable for quality control of medicinal materials and products when the effective components are not fully or not required to be fully identified. Besides examining factors such as the origin, harvesting season, harvested parts, processing methods, and storage time of medicinal materials to provide a basis for identifying the authenticity and quality of raw materials before production, TCM fingerprinting can also be used for quality control during the TCM production process: tracking changes in certain chemical components in preparations, and monitoring the consistency and stability of quality between raw materials and finished products, as well as between different batches of finished products. Compared with quality analysis methods that determine the content of index components, fingerprinting can more comprehensively reflect the types and quantities of chemical components in traditional Chinese medicine. Given that the effective components of traditional Chinese medicine compound preparations have not yet been fully elucidated, fingerprinting can achieve a comprehensive evaluation of the intrinsic quality of traditional Chinese medicine and effective control of its overall substances. It is one of the effective means of quality control of traditional Chinese medicine and its preparations.

[0005] Currently, no literature reports a comprehensive analysis of the chemical composition and fingerprint spectral studies of the traditional Chinese medicine composition containing angelica and cinnamon claimed in this invention. In the prior art, only the quality analysis of a single component in the traditional Chinese medicine compound is performed. There is no comprehensive and systematic quality control method to reflect the quality status of the main reference sample components in the traditional Chinese medicine compound and finished product of this invention. It is impossible to effectively control its production process and product quality, and cannot better guarantee its clinical efficacy. Therefore, it is necessary to adopt a fingerprint spectral quality control method that can comprehensively control the overall quality of the traditional Chinese medicine compound of this invention to control its key quality. Summary of the Invention

[0006] Based on this, the present invention provides a method for constructing a fingerprint spectrum of a traditional Chinese medicine composition containing angelica and cinnamon, the method comprising the following steps:

[0007] Preparation of the test solution of the traditional Chinese medicine composition: Take an appropriate amount of the decoction of the traditional Chinese medicine composition containing Angelica sinensis and cinnamon, place it in a container, add solvent, sonicate for a period of time, make up to volume, shake well, centrifuge, take the supernatant and filter, take the filtrate to obtain the test solution of the traditional Chinese medicine composition; wherein, the traditional Chinese medicine composition includes Angelica sinensis, cinnamon, white peony root, ginger, jujube and licorice.

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

[0009] Based on the results of high-performance liquid chromatography (HPLC) detection of the test solution and the reference solution of the traditional Chinese medicine composition, a fingerprint spectrum of the traditional Chinese medicine composition was obtained.

[0010] The chromatographic conditions for this high-performance liquid chromatography (HPLC) detection are as follows: a column packed with octadecyl-bonded silica gel is used; mobile phase A is selected from one or more of acetonitrile, methanol, and tetrahydrofuran; mobile phase B is an aqueous acid solution, an aqueous alkaline solution, and / or an aqueous buffer 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. 52–58 min, 75% B → 71% B; 58–62 min, 71% B → 68% B; 62–80 min, 68% B → 44% B; 80–83 min, 44% B → 26% B; 83–88 min, 26% B → 15% B; 88–89 min, 15% B → 95% B; 89–100 min, 95% B; flow rate 0.5–1.5 ml / min, column temperature 10–40 °C, detection wavelength 180–350 nm, injection volume 1–20 μl.

[0011] Furthermore, this 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 decoction of this traditional Chinese medicine composition has a mass of 5 to 30g, for example, about 15g.

[0015] Furthermore, the mass-to-volume ratio of the decoction of the traditional Chinese medicine composition to the solvent is 0.4 to 0.8, for example, about 0.6, in 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 duration of the ultrasonic treatment is 20–40 minutes, for example, about 30 minutes.

[0019] Furthermore, the centrifugation 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 ratio of the angelica, cinnamon, white peony root, ginger, jujube and licorice is (45-60):(30-50):(70-90):(30-50):(25-45):(20-40).

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

[0023] Furthermore, the preparation method of the decoction of the traditional 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, adding water to soak them, boiling them to a boil and boiling for a period of time, then keeping them at a gentle boil for a period of time, filtering them with a filter while they are hot to obtain the decoction of the traditional Chinese medicine composition containing angelica and cinnamon.

[0024] Furthermore, the volume / mass ratio of the water to the herbal composition containing angelica and cinnamon is 5 to 10, for example, about 7.

[0025] Furthermore, the container is either a ceramic 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 pot should be 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 at the simmering stage is 500–1500W, for example, about 900W.

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

[0032] Furthermore, the volume-to-volume ratio of the decoction containing angelica and cinnamon to the water is 0.1 to 0.5, for example, 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 content in the reference solution is 20–40 μg / ml, for example, about 30 μg / ml.

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

[0040] Furthermore, the cinnamic acid content 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 for this high-performance liquid chromatography 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] Furthermore, between the 0th minute and the 77th minute, the detection wavelength is 210–280 nm, for example, about 250 nm.

[0045] Furthermore, between the 77th and 100th minutes, 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 this ferulic acid is not less than 5000.

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

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

[0050] Furthermore, mobile phase A is acetonitrile.

[0051] Furthermore, the aqueous acid solution, aqueous alkaline solution, and / or aqueous buffer salt solution are selected from one or more weak acids and their salts, weak bases and their salts of different concentrations.

[0052] Furthermore, the aqueous acid solution, aqueous alkaline solution, and / or aqueous buffer solution are selected from different concentrations of 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.

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

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

[0055] Furthermore, the acidic aqueous solution is an aqueous solution of approximately 0.1% phosphoric acid.

[0056] Furthermore, the buffer salt solution is an acetate solution and / or an acetate solution.

[0057] Furthermore, the pH value of this buffer salt solution is no greater than 7.0.

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

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

[0060] Furthermore, when the detection wavelength is 250 nm between 0 minutes and 77 minutes, and 210 nm between 77 minutes and 100 minutes, 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] Furthermore, when the detection wavelength is 250nm between the 0th and 77th minutes, and 210nm between the 77th and 100th minutes, peak 1 originates from white peony root, peak 2 originates from white peony root, peak 3 originates from white peony root, peak 4 originates from white peony root, peak 5 originates from angelica root, peak 6 originates from stir-fried licorice root, peak 7 originates from white peony root, peak 8 originates from cinnamon root, peak 9 originates from cinnamon root, peak 10 originates from stir-fried licorice root, peak 11 originates from stir-fried licorice root, peak 12 originates from stir-fried licorice root, peak 13 originates from ginger root, and peak 14 originates from angelica root.

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

[0063] (1) Establish a standard fingerprint spectrum of the reference sample of traditional Chinese medicine composition according to the above fingerprint spectrum construction method;

[0064] (2) Take the test solution of the traditional Chinese medicine composition and perform detection according to the chromatographic conditions in the above fingerprint chromatogram construction method to obtain the fingerprint chromatogram of the traditional Chinese medicine composition sample; and

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

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

[0067] (1) The fingerprint spectrum of the sample to be tested of the traditional Chinese medicine composition showed 14 characteristic chromatographic peaks, and the retention time of each characteristic chromatographic peak was within ±10% of the retention time value of the corresponding reference chromatographic peak in the standard fingerprint spectrum of the reference sample of the traditional Chinese medicine composition.

[0068] (2) Taking peak 4 as peak S, the relative retention times of each characteristic chromatographic peak in the fingerprint chromatogram of the Chinese herbal composition sample to be tested and peak S are within ±10% of the relative retention times of each characteristic chromatographic peak in the standard fingerprint chromatogram of the Chinese herbal composition reference sample; and

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

[0070] According to another aspect of the present invention, the above-described construction method or the above-described quality control method is provided for use in the quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia.

[0071] The beneficial effects of this invention are:

[0072] The beneficial effects of this invention are:

[0073] In summary, this invention provides a method for fingerprint analysis and quality control of a reference sample containing angelica and cinnamon. This method identifies 14 common characteristic peaks, is simple to implement, and has a short analysis time. It solves the problems of difficult separation of fingerprint characteristic peaks and interference from impurity peaks, ensuring the chemical composition stability and safety of the reference sample. This provides an important reference for the quality control of subsequent preparations, ensuring product quality stability, guaranteeing the efficacy of the traditional Chinese medicine compound, and enabling the compound to better serve human health.

[0074] Specifically, compared with the prior art, the beneficial effects of the present invention are as follows:

[0075] (1) The fingerprint spectrum of the reference sample of the traditional Chinese medicine compound of the present invention is established, which overcomes the defect that the content determination of a single component is difficult to reflect the overall content. It can control the intrinsic quality of the reference sample of the traditional Chinese medicine compound of the present invention from an overall and macroscopic perspective, ensuring the efficacy of the drug. By using the main means of modern drug research, the classics can be inherited and the classic prescriptions can be more formally controlled.

[0076] (2) The chemical composition of the traditional Chinese medicine compound reference sample of the present invention is complex, and it is difficult to separate its characteristic peaks. In the process of establishing the fingerprint spectrum, the present invention adopts the gradient elution method to solve the problem of fingerprint characteristic peaks being difficult to separate and the 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 identified, and their relative retention time, relative peak area and similarity were studied, which ensured the chemical composition stability and safety of the reference sample, and provided important reference and quality reference for the subsequent quality control of compound preparations.

[0078] (4) The fingerprint spectrum of each effective component in the traditional Chinese medicine compound reference sample of the present invention is regarded as a whole. The order and interrelationship of each characteristic peak are emphasized. This 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. It also reduces the possibility of artificial processing to meet the quality standards. This provides 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, and is convenient and easy to master. Attached Figure Description

[0080] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without exceeding the scope of protection claimed by the present invention.

[0081] Figure 1 This is a liquid chromatography fingerprint of a granular reference sample of a traditional Chinese medicine composition containing angelica and cinnamon according to the present invention. Detailed Implementation

[0082] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0083] Unless otherwise stated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by one of ordinary skill in the field of this invention or the field of application of such terms. While any methods, conditions, substances, or materials similar to or equivalent to those disclosed herein may be used in the practice of this invention, preferred methods, conditions, substances, or materials are described herein.

[0084] This invention is intended to cover all options, variations, and equivalents that may be included in the field of prior art as defined in the claims. Those skilled in the art will recognize many similar or equivalent methods and substances described herein that can be applied in the practice of this invention. This invention is by no means limited to the description of methods and substances.

[0085] The singular forms “a,” “an,” and “the” used in the specification and appended claims include plural indicators unless the context clearly specifies otherwise.

[0086] In this invention, the term "comprising" and "including" are synonymous. The terms "comprising," "including," "having," "containing," or any other variations thereof as used herein are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such a composition, step, method, article, or apparatus.

[0087] As described in the background section, no literature reports a comprehensive analysis of the chemical composition and fingerprinting of the traditional Chinese medicine composition containing Angelica sinensis and cinnamon claimed in this invention. To address the above problems, this invention provides a method for constructing a fingerprint of a traditional Chinese medicine composition containing Angelica sinensis and cinnamon, the method comprising the following steps:

[0088] Preparation of the test solution of the traditional Chinese medicine composition: Take an appropriate amount of the decoction of the traditional Chinese medicine composition containing Angelica sinensis and cinnamon, place it in a container, add solvent, sonicate for a period of time, make up to volume, shake well, centrifuge, take the supernatant and filter, take the filtrate to obtain the test solution of the traditional Chinese medicine composition; wherein, the traditional Chinese medicine composition includes Angelica sinensis, cinnamon, white peony root, ginger, jujube and licorice.

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

[0090] Based on the results of high-performance liquid chromatography (HPLC) detection of the test solution and the reference solution of the traditional Chinese medicine composition, a fingerprint spectrum of the traditional Chinese medicine composition was obtained.

[0091] The chromatographic conditions for this high-performance liquid chromatography (HPLC) detection are as follows: a column packed with octadecyl-bonded silica gel is used; mobile phase A is selected from one or more of acetonitrile, methanol, and tetrahydrofuran; mobile phase B is an aqueous acid solution, an aqueous alkaline solution, and / or an aqueous buffer 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. 52–58 min, 75% B → 71% B; 58–62 min, 71% B → 68% B; 62–80 min, 68% B → 44% B; 80–83 min, 44% B → 26% B; 83–88 min, 26% B → 15% B; 88–89 min, 15% B → 95% B; 89–100 min, 95% B; flow rate 0.5–1.5 ml / min, column temperature 10–40 °C, detection wavelength 180–350 nm, injection volume 1–20 μl.

[0092] In this 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 a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range "0.5 to 1.5" is disclosed, the described range should be interpreted as including ranges such as "0.5 to 1.5", "0.5 to 1.3", "0.5 to 1.1", "0.5 to 0.9", "0.5 to 0.7", "0.7 to 1.5", "0.7 to 1.3", "0.7 to 1.1", "0.7 to 0.9", "0.9 to 1.5", "0.9 to 1.3", "0.9 to 1.1", "1.1 to 1.5", "1.1 to 1.3", "1.3 to 1.5", etc. When a range of values ​​is described in this document, unless otherwise stated, the range is intended to include its 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, the solvent in the preparation of the test solution of the traditional Chinese medicine composition is an alcohol, such as methanol.

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

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

[0097] In this 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 of the decoction of the traditional Chinese medicine composition to the solvent is 0.4 to 0.8, for example, about 0.6, in g / ml.

[0099] In this 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 to 600 W, for example, about 500 W.

[0101] In this 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, for example, about 40 kHz.

[0103] In this 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, for example, about 30 minutes.

[0105] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 30" includes 30 ±5%, or from 28.5 to 31.5.

[0106] In a preferred embodiment, the centrifugation speed is 4000 to 6000 rpm / min, for example, about 5000 rpm / min.

[0107] In this invention, "about" refers to a value within a 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 minutes, for example, about 10 minutes.

[0109] In this 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, white peony root, ginger, jujube and licorice is (45-60):(30-50):(70-90):(30-50):(25-45):(20-40).

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

[0112] In this 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 method for preparing the decoction of the traditional 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, adding water to soak them, boiling them to a boil and boiling for a period of time, then keeping them at a gentle boil for a period of time, filtering them with a filter while they are hot to obtain the decoction of the traditional Chinese medicine composition containing angelica and cinnamon.

[0114] In a preferred embodiment, the volume / mass ratio of the water to the herbal composition containing angelica and cinnamon is 5 to 10, for example, about 7.

[0115] In this 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 decoction pot or an electric ceramic stove.

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

[0118] In this 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 pot is covered when it boils.

[0120] In a preferred embodiment, the container has a power of 1000 to 3000W when boiling, for example, about 2000W.

[0121] In this 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, for example, about 10 minutes.

[0123] In this 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 at the simmering stage is 500 to 1500 W, for example, about 900 W.

[0125] In this 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 simmering time is 10 to 100 minutes, for example, about 60 minutes.

[0127] In this 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-to-volume ratio of the decoction containing angelica and cinnamon to the water is 0.1 to 0.5, for example, about 0.3.

[0129] In this 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, for example, about 18 μg / ml.

[0132] In this 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, for example, about 13 μg / ml.

[0134] In this 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 to 10 μg / ml, for example, about 6 μg / ml.

[0136] In this 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 this 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 reference solution contains 20–40 μg / ml of gallic acid, for example, about 30 μg / ml.

[0140] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 30" includes 30 ±5%, or from 28.5 to 31.5.

[0141] In a preferred embodiment, the reference solution contains 50–70 μg / ml of paeoniflorin, for example, about 60 μg / ml.

[0142] In this 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 reference solution contains 40–60 μg / ml of cinnamic acid, for example, about 50 μg / ml.

[0144] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes 50 ± 5%, or from 47.5 to 52.5.

[0145] In a preferred embodiment, the reference solution contains 40–60 μg / ml of glycyrrhizic acid, for example, about 50 μg / ml.

[0146] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes 50 ± 5%, or from 47.5 to 52.5.

[0147] In a preferred embodiment, the flow rate of the high-performance liquid chromatography (HPLC) detection is 0.8 to 1.2 ml / min, for example, about 1.0 ml / min.

[0148] In this 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, for example, about 25°C.

[0150] In this 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, the detection wavelength is 210–280 nm, for example, about 250 nm, between the 0th minute and the 77th minute.

[0152] In this 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, the detection wavelength is 190–230 nm, for example, about 210 nm, between the 77th minute and the 100th minute.

[0154] In this 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 to 15 μl, for example, about 10 μl.

[0156] In this 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 column.

[0159] In a preferred embodiment, the chromatographic column has the following specifications: column length 250 mm, inner diameter 4.6 mm, and particle size 5 μm.

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

[0161] In a preferred embodiment, the acid aqueous solution, alkaline aqueous solution, and / or buffer salt aqueous solution are selected from one or more weak acids and their salts, and weak bases and their salts of different concentrations.

[0162] In a preferred embodiment, the acidic aqueous solution, alkaline aqueous solution, and / or buffer salt aqueous 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.

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

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

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

[0166] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.1%" includes 0.1% ± 5%, or from 0.095% to 0.105%.

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

[0168] In a preferred embodiment, the pH value of the buffer saline solution is not greater than 7.0.

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

[0170] In this 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. "Approximately 38.938" includes ±5% of 38.938, or from 36.9911 to 40.8849; "Approximately 42.241" includes ±5% of 42.241, or from 40.12895 to 44.35305; "Approximately 45.390" includes ±5% of 45.390, or from 43.1205 to 47.6595; "Approximately 64.056" includes ±5% of 64.056, or from 60.8532 to 67.2588; "Approximately 65.959" includes ±5% of 65.959, or from 62.66105 to 69.25695; "Approximately 69.988" includes ±5% of 69.988, or from 66.4886 to 73.4874; "Approximately 72.691" includes ±5% of 72.691, or From 69.05645 to 76.32555; “about 74.593” includes 74.593 ± 5%, or from 70.86335 to 78.32265; “about 78.837” includes 78.837 ± 5%, or from 74.89515 to 82.77885; “about 85.261” includes 85.261 ± 5%, or from 80.99795 to 89.52405.

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

[0172] In this 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 2.203 ± 5%, or from 2.09285 to 2.31315; “about 2.337” includes 2.337 ± 5%, or from 2.22015 to 2.45385; “about 2.427” includes 2.427 ± 5%, or from 2.30565 to 2.54835; “about 2.491” includes 2.491 ± 5%, or from 2.36645 to 2.61555; “about 2.633” includes 2.633 ± 5%, or from 2.50135 to 2.76465; “about 2.847” includes 2.847 ± 5%, or from 2.70465 to 2.98935.

[0173] In a preferred embodiment, when the detection wavelength is 250 nm between the 0th and 77th minutes and 210 nm between the 77th and 100th minutes, 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 is 250 nm between the 0th and 77th minutes and 210 nm between the 77th and 100th minutes, peak 1 originates from white peony root, peak 2 originates from white peony root, peak 3 originates from white peony root, peak 4 originates from white peony root, peak 5 originates from angelica root, peak 6 originates from stir-fried licorice root, peak 7 originates from white peony root, peak 8 originates from cinnamon root, peak 9 originates from cinnamon root, peak 10 originates from stir-fried licorice root, peak 11 originates from stir-fried licorice root, peak 12 originates from stir-fried licorice root, peak 13 originates from ginger root, and peak 14 originates from angelica root.

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

[0176] (1) Establish a standard fingerprint spectrum of the reference sample of traditional Chinese medicine composition according to the above fingerprint spectrum construction method;

[0177] (2) Take the test solution of the traditional Chinese medicine composition and perform detection according to the chromatographic conditions in the above fingerprint chromatogram construction method to obtain the fingerprint chromatogram of the traditional Chinese medicine composition sample; and

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

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

[0180] (1) The fingerprint spectrum of the sample to be tested of the traditional Chinese medicine composition showed 14 characteristic chromatographic peaks, and the retention time of each characteristic chromatographic peak was within ±10% of the retention time value of the corresponding reference chromatographic peak in the standard fingerprint spectrum of the reference sample of the traditional Chinese medicine composition.

[0181] (2) Taking peak 4 as peak S, the relative retention times of each characteristic chromatographic peak in the fingerprint chromatogram of the Chinese herbal composition sample to be tested and peak S are within ±10% of the relative retention times of each characteristic chromatographic peak in the standard fingerprint chromatogram of the Chinese herbal composition reference sample; and

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

[0183] According to another aspect of the present invention, the above-described construction method or the above-described quality control method is provided for use in the quality detection and / or quality evaluation and / or quality control of a traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia.

[0184] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or conditions recommended by the manufacturer.

[0185] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0186] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this patent specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0187] Example

[0188] Example

[0189] 1. Instruments and reagents

[0190] 1.1 Instruments

[0191] High-performance liquid chromatograph, model: Agilent 1290_DAD HPLC (Agilent Technologies); High-performance liquid chromatograph, model: Waters 2695_2996 (Waters Technologies); 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 Measurement Instrument Co., Ltd.); Benchtop 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 Chemicals

[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 National Institutes for Food and Drug Control); cinnamaldehyde (batch number: 110710-202223, content: 98.9%, China National Institutes for Food and Drug Control); ligustilide (batch number: 111737-202311, content: 100.0%, China National Institutes for Food and Drug Control); 6-gingerol (batch number: 111833-202007, content: 99.3%). (China National Institutes for Food and Drug Control); Gallic acid (batch number: 110831-201906, content: 91.5%, China National Institutes for Food and Drug Control); Ammonium glycyrrhizate (batch number: 110731-202122, content: 94.4%, China National Institutes for Food and Drug Control); Cinnamic acid (batch number: 110786-202305, content: 99.8%, China National Institutes for Food and Drug Control); Paeoniflorin (batch number: 110736-202447, content: 98.1%, China National Institutes for Food and Drug Control).

[0194] 1.3 Sample Information

[0195] The present invention includes the following reference samples: a granule standard sample of a traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia (batch number: 2024062801-L, source: self-made); a granule standard sample of a traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia (batch number: 2024072501-L, source: self-made); a granule standard sample of a traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia (batch number: 2024082904-L, source: self-made); and a negative reference sample of a granule composition containing Angelica sinensis and Cinnamomum cassia lacking Angelica sinensis (batch number: 202409). 0201-L (Source: Self-made); The traditional Chinese medicine composition granules containing Angelica sinensis and cinnamon of the present invention, lacking cinnamon, negative reference sample (batch number: 2024090204-L, source: self-made); The traditional Chinese medicine composition granules containing Angelica sinensis and cinnamon of the present invention, lacking white peony root, negative reference sample (batch number: 2024090202-L, source: self-made); The traditional Chinese medicine composition granules containing Angelica sinensis and cinnamon of the present invention, lacking ginger, negative reference sample (batch number: 2024090301 ... white peony root, negative reference sample (batch number: 2024090202-L, source: self-made); The following are sample references for traditional Chinese medicine compositions containing cinnamon and angelica: granules lacking jujube (batch number: 2024090302-L, source: self-made); granules containing angelica and cinnamon of the present invention lacking roasted licorice (batch number: 2024090203-L, source: self-made); angelica slices (batch number: ZA00323102101-01, origin: Minxian County, Dingxi City, Gansu Province, source: self-made); cinnamon slices (batch number: ZA04223112801, origin: Pingxiang County, Guigang City, Guangxi Zhuang Autonomous Region, source: self-made). Homemade); White peony root slices (batch number: ZA00123121502, place of origin: Qiaocheng District, Bozhou City, Anhui Province, source: homemade); Jujube slices (batch number: ZA00223121402, place of origin: Ningyang County, Tai'an City, Shandong Province, source: homemade); Ginger slices (batch number: ZA00524071501, place of origin: Wanyuan County, Dazhou City, Sichuan Province, source: homemade); Fried licorice root slices (batch number: ZD00423090401-01, place of origin: Hangjin Banner, Ordos City, Inner Mongolia, source: homemade).

[0196] 1.4 Solution Preparation

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

[0198] Take 55.20g of Angelica sinensis, 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 root. Grind the above six ingredients into coarse granules, place them in a ceramic decoction pot, add 2000ml of water, cover, and soak for 45 minutes. Boil on an electric ceramic stove at 2000W for about 10 minutes, then simmer at 900W until reduced to 600ml (about 60 minutes). Filter the decoction through a 200-mesh nylon filter while hot to obtain the final product.

[0199] (2) Preparation of the reference solution:

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

[0201] (3) Preparation of the test solution

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

[0203] (4) Preparation of negative control solution

[0204] Preparation of negative sample solution for Angelica sinensis deficiency: Weigh the other herbs for Angelica sinensis deficiency according to the prescription, prepare negative sample for Angelica sinensis deficiency according to the preparation method of the reference sample, and prepare negative sample solution for Angelica sinensis deficiency according to item "(3)".

[0205] Negative control solutions lacking Angelica sinensis, cinnamon, white peony root, ginger, jujube, and stir-fried licorice were prepared using the same method.

[0206] (5) Blank solvent: methanol.

[0207] 3. Establishment of Feature Mapping Method

[0208] 3.1 Study on chromatographic conditions of characteristic spectra

[0209] The proposed initial chromatographic conditions are as follows:

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

[0211] Chromatographic conditions and system suitability tests were performed using octadecyl-bonded silica gel as the stationary phase (Agilent 5TC-C18 4.6×250mm 5μm); acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, with gradient elution performed according to the specifications in Table 1; flow rate was 1 ml per minute; column temperature was 25℃; and full wavelength scanning was performed.

[0212] Table 1. Characteristic chromatograms of the traditional Chinese medicine composition containing Angelica sinensis and Cinnamomum cassia of the present invention, chromatographic condition gradient 1

[0213]

[0214]

[0215] Preparation of reference solution: Accurately weigh appropriate amounts of ferulic acid, paeoniflorin, cinnamaldehyde, 6-gingerol, ligustilide, gallic acid, ammonium glycyrrhizate, cinnamic acid, and paeoniflorin reference standards, and add methanol to prepare a mixed solution containing 18 μg ferulic acid, 1.0 mg paeoniflorin, 13 μg cinnamaldehyde, 6 μg 6-gingerol, 18 μg ligustilide, 30 μg gallic acid, 50 μg ammonium glycyrrhizate, 50 μg cinnamic acid, and 60 μg paeoniflorin per ml, which will be used as the reference solution.

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

[0217] The assay involves precisely pipetting 10 μl of the reference solution and the test solution, injecting them into the liquid chromatograph, and measuring the results.

[0218] Table 2 summarizes the optimal absorption wavelengths of the reference solutions.

[0219] Table 2 Summary of Optimal Absorption Wavelengths for Reference Solutions

[0220] Reference solution Peak time Optimal absorption wavelength (nm) gallic acid 7.531 216、272 paeoniflorin 28.460 232 ferulic acid 34.661 218、236、322 Ligusticum 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 showed that, combined with the full-wavelength UV absorption spectrum, UV absorption fluorescence was observed at wavelengths of 200 nm to 300 nm. Comparing the liquid phase spectra at 210 nm, 250 nm, and 254 nm, the response values ​​of the characteristic peaks at 254 nm and 250 nm were significantly better than those at 210 nm within the 0–75 minute range. 6-Gingerol (t) was visible at 210 nm within the 75–100 minute range. R =77.321) and ligustilide (t R =84.083); therefore, the detection wavelength of the characteristic spectrum of the granular reference sample of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention is determined to be: 250 nm for the detection wavelength within 0 to 77 minutes, and 210 nm for the detection wavelength within 77 to 100 minutes. Given the poor separation of the characteristic peaks within 30 to 40 minutes, it is further optimized to gradient 2 in 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 showed that the resolution and response values ​​of each characteristic peak under gradient 2 met the requirements of the liquid chromatography characteristic spectrum. Therefore, this gradient was determined as the characteristic spectrum method for the reference sample of the traditional Chinese medicine composition granules containing Angelica sinensis and Cinnamomum cassia of this invention. The detection wavelengths were: 250 nm for 0–77 minutes and 210 nm for 77–100 minutes. The chromatogram of the reference sample of the decoction of the traditional Chinese medicine composition granules containing Angelica sinensis and Cinnamomum cassia of this invention showed 21 chromatographic peaks. Considering that the peak area response values ​​of peaks 7, 9–12, and 14 were low, and the peak shape of peak 18 was poor, considering batch-to-batch differences, they were not included in the characteristic peak standard of the characteristic spectrum. The response values ​​and resolution of the remaining chromatographic peaks were good. Therefore, a total of 14 characteristic peaks were included in the characteristic spectrum of the reference sample of the decoction of the traditional Chinese medicine composition granules containing Angelica sinensis and Cinnamomum cassia of this invention. The elution time of peak 21 (ligustilide) was about 85 minutes, which had returned to the initial gradient. Further optimization was carried out to gradient 3 as shown in Table 4.

[0225] Table 4 Gradient 3

[0226]

[0227]

[0228] The results show that the separation degree and response value of each characteristic peak under this gradient have met the requirements of liquid phase characteristic spectrum. Therefore, this gradient is determined as the characteristic spectrum method of the granular reference sample of the traditional Chinese medicine composition containing angelica and cinnamon in this invention. The detection wavelength is: 250 nm for 0 to 77 minutes and 210 nm for 77 to 100 minutes.

[0229] 3.2. Investigation on the preparation of reference sample test solution

[0230] Based on the results of the study on the preparation process of the reference sample, which is preserved in the form of decoction, the preparation method of the decoction test solution was investigated. Therefore, it was determined that 15g of decoction would be used for the study.

[0231] 3.2.1 Investigation of Extraction Solvents

[0232] Six portions of the decoction were taken, and the effects of pure water, 50% methanol, and methanol as extraction solvents on the characteristic chromatograms of the granular reference samples of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention were investigated.

[0233] Take 6 portions of the standard sample decoction, each about 15g, accurately weigh them, place them in 25ml volumetric flasks, add appropriate amounts of pure water, 50% methanol, and methanol respectively, sonicate (500W, 40kHz) for 30 minutes, cool, make up to volume, shake well, centrifuge at 5000rpm / min for 10min, take the supernatant and filter it, take the filtrate to obtain the final product.

[0234] The results showed that the number of characteristic peaks of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention was the same after different extraction solvents were used for treatment. The total peak area was slightly larger after methanol treatment, so methanol was the appropriate solvent for treatment.

[0235] 3.2.2, Examination of extraction time

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

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

[0238] The results showed that the number of characteristic peaks and the total peak area were similar after different extraction times for the traditional Chinese medicine composition granules containing angelica and cinnamon of the present invention. Considering the continuity of the experiment, the extraction time was 30 minutes.

[0239] 3.2.3 Sampling Quantity Examination

[0240] Six portions of the decoction were taken, and the effects of sample amounts of 10g, 15g, and 20g on the characteristic spectrum of the granular reference sample of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention were investigated.

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

[0242] The results showed that after methanol treatment, the total peak area of ​​the herbal composition containing angelica and cinnamon granules of the present invention varied proportionally with different sample amounts (10–20 g), indicating that the detection method was stable within the sample amount range of 10–20 g. Considering that the same method was used for both content detection and characteristic chromatographic detection, the sample amount was determined to be 15 g.

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

[0244] 4. Methodological Validation

[0245] 4.1 Specificity

[0246] The blank solvent, reference solution, the granular reference sample solution of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention, the negative control solution of each herb, and the single herb control solution are injected for analysis. The blank solvent should not interfere with the determination of the elution position of the characteristic peak. In addition, the assignment of the chromatographic peaks should be determined based on the detection results of the negative control solution and the single herb control solution. Figure 1 This is a liquid chromatography fingerprint of a granular reference sample of a traditional Chinese medicine composition containing angelica and cinnamon according to the present invention.

[0247] In summary, the chromatographic peak assignment results of the characteristic chromatogram of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention show that the chromatogram of the decoction of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention mainly presents 14 chromatographic peaks. Peaks 1-4 and peak 7 belong to white peony root, with peak 1 being gallic acid and peak 4 being paeoniflorin; peaks 5 and 14 belong to angelica root, which are ferulic acid and ligustilide, respectively; peaks 6 and 10-12 belong to stir-fried licorice root, with peak 12 being glycyrrhizic acid; peaks 8 and 9 belong to cinnamon root, with peak 8 being cinnamic acid and peak 9 being cinnamaldehyde; and peak 13 belongs to ginger root, which is 6-gingerol.

[0248] Blank solvent, reference solution, the granular reference sample solution of the traditional Chinese medicine composition containing angelica and cinnamon of the present invention, negative control solutions of each herb, and single-herb control solutions were injected for analysis. At the determined characteristic peak positions, the blank solvent and negative control solutions showed no interference. Details of the specific chromatographic peaks and their corresponding herbs in the chromatograms are shown in Table 5.

[0249] Table 5. Details of the specific chromatographic peaks in the chromatograms and their corresponding medicinal ingredients.

[0250] medicinal smell Angelica sinensis Cinnamon White peony root Ginger jujube Stir-fried licorice Remark Belonging 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 sinensis Peak 6 - - - - - + / Stir-fried licorice Peak 7 - - + - - - / White peony root Peak 8 - + - - - - Cinnamon acid Cinnamon Peak 9 - + - - - - Cinnamaldehyde Cinnamon Peak 10 - - - - - + / Stir-fried licorice Peak 11 - - - - - + / Stir-fried licorice Peak 12 - - - - - + glycyrrhizic acid Stir-fried licorice Peak 13 - - - + - - 6-Gingerol Ginger Peak 14 + - - - - - ligustilide Angelica sinensis

[0251] 4.2 Precision

[0252] 4.2.1 Repeatability

[0253] Prepare 6 test solutions according to the above test solution preparation method, inject and detect them, record the chromatograms, and calculate the RSD value of the relative retention time of the characteristic peak and the S peak (paeoniflorin); the RSD of the relative retention time is ≤3%.

[0254] The results show that the RSD of the relative retention time of each characteristic peak in the repeatability experiment is ≤3%, indicating that the method has good repeatability.

[0255] 4.2.2 Intermediate Precision

[0256] Two personnel prepared six test solutions according to the above-determined conditions, and injected them into an Agilent high-performance liquid chromatograph and a Waters high-performance liquid chromatograph for detection. The RSD value of the relative retention time of each characteristic peak and the S peak (paeoniflorin) was calculated. If the RSD of the relative retention time is ≤3%, it indicates that the intermediate precision is good.

[0257] The results show that the RSD of the relative retention times of each characteristic peak in the intermediate precision tests for Waters and Agilent is ≤3%. This indicates that the method has good intermediate precision in both Waters and Agilent.

[0258] 4.2.3 Instrument Precision

[0259] Prepare a test solution under the conditions defined above, inject it six times consecutively, record the chromatogram, and calculate the RSD values ​​of the retention time and peak area of ​​the characteristic peak and the S peak (paeoniflorin). If the RSD of the relative retention time is ≤3% and the RSD of the relative peak area is ≤5%, the relative peak area can be appropriately relaxed for peaks with a smaller proportion of the total peak area. This indicates good instrument precision.

[0260] The results showed that the RSD of the relative retention time of each characteristic peak in the instrument precision experiment was ≤3% and 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 test solution according to the above test 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 the sample for detection, 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 indicates that the flow rate is robust.

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

[0265] 4.3.2 Column Temperature Durability

[0266] Prepare one sample solution according to the above preparation method. Inject the sample and detect it 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 indicates that the flow rate robustness is good.

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

[0268] 4.3.3 Durability under different acid concentrations

[0269] Prepare one test solution according to the above test solution preparation method, and inject it into 0.09%, 0.10%, and 0.11% phosphoric acid solution for detection. Record the chromatograms 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 indicates that the robustness to different acid concentrations is good.

[0270] The results showed that the relative average deviation of the relative retention time of each characteristic peak in the experiment with different acid concentrations was ≤3%, indicating that the performance was good with different acid concentrations.

[0271] 4.3.4 Durability of Different Chromatographic Columns

[0272] Prepare one test solution according to the above test solution preparation method, use three different numbered chromatographic columns, record the chromatograms, 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 indicates that the different chromatographic columns have good durability.

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

[0274] 4.4 Stability of the test solution

[0275] Prepare one test solution according to the above method, and inject it for detection at 0, 3, 6, 10, 15, 18, 24, 36, 48, and 62 hours. Record the chromatograms and calculate the RSD values ​​of the relative retention times and relative peak areas of each characteristic peak. The relative peak area of ​​peaks with a smaller proportion of the total peak area can be appropriately relaxed. This indicates that the test solution has good stability.

[0276] The results showed 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 was ≤3%; the RSD of the relative peak area was ≤5%, except for peak 9. Since the total peak area of ​​peak 9 accounted for ≤1.0%, this limitation could be relaxed. This indicates that the test solution has good stability within 62 hours.

[0277] 5. Validation of the characteristic spectrum method for reference samples

[0278] Based on the above research results and in accordance with the analytical method validation guidelines 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. Therefore, this detection method is of good quality, as follows.

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

[0280] Chromatographic conditions and system suitability tests were performed using octadecyl-bonded silica gel as the stationary phase (Agilent 5TC-C18 4.6×250mm 5μm); acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, with gradient elution as specified in Table 4; flow rate of 1 mL / min; column temperature of 25℃; detection wavelength of 250 nm for 0–77 minutes and 210 nm for 77–100 minutes. The theoretical plate number, calculated based on the ferulic acid peak, should be no less than 5000.

[0281] Preparation of reference solution: Accurately weigh appropriate amounts of ferulic acid, cinnamaldehyde, 6-gingerol, ligustilide, gallic acid, ammonium glycyrrhizate, cinnamic acid, and paeoniflorin reference standards, and add methanol to prepare a mixed solution containing 18 μg ferulic acid, 13 μg cinnamaldehyde, 6 μg 6-gingerol, 18 μg ligustilide, 30 μg gallic acid, 50 μg ammonium glycyrrhizate, 50 μg cinnamic acid, and 60 μg paeoniflorin per ml, which will be used as the reference solution.

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

[0283] The assay involves precisely pipetting 10 μl of the reference solution and the test solution, injecting them into the liquid chromatograph, and measuring the results.

[0284] The embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present invention. The descriptions of the embodiments above are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, any changes or modifications made by those skilled in the art based on the ideas of the present invention, its specific implementation methods, and its application scope, are all within the scope of protection of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A method for constructing a fingerprint spectrum of a traditional Chinese medicine composition containing angelica and cinnamon, characterized in that, The construction method includes the following steps: Preparation of the test solution of the traditional Chinese medicine composition: Take an appropriate amount of the decoction of the traditional Chinese medicine composition containing angelica and cinnamon, place it in a container, add methanol, sonicate for a period of time, make up to volume, shake well, centrifuge, take the supernatant and filter, take the filtrate to obtain the test solution of the traditional Chinese medicine composition; wherein, the traditional Chinese medicine composition includes angelica, cinnamon, white peony root, ginger, jujube and licorice. Preparation of reference solutions: Weigh appropriate amounts of gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide reference standards, and add methanol to prepare reference solutions with concentrations of 1–100 μg / ml for gallic acid, paeoniflorin, ferulic acid, cinnamic acid, cinnamaldehyde, glycyrrhizic acid, 6-gingerol, and ligustilide, respectively. Based on the results of high-performance liquid chromatography (HPLC) detection of the test solution and the reference solution of the traditional Chinese medicine composition, a fingerprint spectrum of the traditional Chinese medicine composition was obtained. The chromatographic conditions for high-performance liquid chromatography (HPLC) detection were as follows: a column packed with octadecyl-bonded silica gel was used; mobile phase A was acetonitrile; mobile phase B was approximately 0.1% aqueous phosphoric acid solution; and the gradient elution program was 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, 80% → 75% B; 52–58 min… The flow rate was approximately 1 ml / min, the column temperature was approximately 25°C, the detection wavelength was approximately 250 nm from 0 to 77 minutes, the detection wavelength was approximately 210 nm from 77 to 100 minutes, and the injection volume was 1 to 20 μl. The flow rate was 75% B → 71% B; 58–62 min, 71% B → 68% B; 62–80 min, 68% B → 44% B; 80–83 min, 44% B → 26% B; 83–88 min, 26% B → 15% B; 88–89 min, 15% B → 95% B; 89–100 min, 95% B. The specifications of the chromatographic column are as follows: column length 250 mm, inner diameter 4.6 mm, particle size 5 μm; The fingerprint spectrum includes peaks 1-14, with peak 4 serving as a reference peak. The average relative retention times of peaks 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14 are approximately 0.299, 0.710, 0.918, 1.300, 1.411, 1.516, 2.139, 2.203, 2.337, 2.427, 2.491, 2.633, and 2.847, respectively.

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

3. The construction method according to claim 1, characterized in that, In the preparation of the test solution of the traditional Chinese medicine composition, the container is a volumetric flask.

4. The construction method according to claim 1, characterized in that, The decoction of the traditional Chinese medicine composition has a mass of 5-30g.

5. The construction method according to claim 4, characterized in that, The decoction of the traditional Chinese medicine composition weighs approximately 15g.

6. The construction method according to claim 1, characterized in that, The mass-to-volume ratio of the decoction of the traditional Chinese medicine composition to the methanol is 0.4 to 0.8, with units of g / ml.

7. The construction method according to claim 6, characterized in that, The mass-to-volume ratio of the decoction of the traditional Chinese medicine composition to the methanol is approximately 0.6, expressed in g / ml.

8. The construction method according to claim 1, characterized in that, The power of the ultrasonic treatment is 400-600W.

9. The construction method according to claim 8, characterized in that, The power of the ultrasonic treatment is approximately 500W.

10. The construction method according to claim 1, characterized in that, The frequency of the ultrasonic treatment is 30–50 kHz.

11. The construction method according to claim 10, characterized in that, The frequency of the ultrasonic treatment is approximately 40 kHz.

12. The construction method according to claim 1, characterized in that, The ultrasonic treatment time is 20 to 40 minutes.

13. The construction method according to claim 12, characterized in that, The ultrasonic treatment lasts for approximately 30 minutes.

14. The construction method according to claim 1, characterized in that, The centrifuge speed is 4000-6000 rpm / min.

15. The construction method according to claim 14, characterized in that, The centrifuge speed is approximately 5000 rpm / min.

16. The construction method according to claim 1, characterized in that, The centrifugation time is 5 to 15 minutes.

17. The construction method according to claim 16, characterized in that, The centrifugation time is approximately 10 minutes.

18. The construction method according to claim 1, characterized in that, The mass ratio of the Angelica sinensis, the cinnamon, the white peony root, the ginger, the jujube and the licorice is (45-60):(30-50):(70-90):(30-50):(25-45):(20-40).

19. The construction method according to claim 18, characterized in that, The mass ratio of the angelica, cinnamon, white peony root, ginger, jujube and licorice is approximately 55.2: approximately 41.4: approximately 82.8: approximately 41.4: approximately 36: approximately 27.

6.

20. The construction method according to claim 1, characterized in that, The preparation method of the decoction of the traditional 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, adding water to soak them, boiling them to a boil and boiling for a period of time, then keeping them at a simmer for a period of time, filtering them with a filter while they are hot to obtain the decoction of the traditional Chinese medicine composition containing angelica and cinnamon.

21. The construction method according to claim 20, characterized in that, The volume / mass ratio of the water to the traditional Chinese medicine composition containing angelica and cinnamon is 5 to 10.

22. The construction method according to claim 21, characterized in that, The volume / mass ratio of the water to the traditional Chinese medicine composition containing angelica and cinnamon is approximately 7.

23. The construction method according to claim 20, characterized in that, The container is a ceramic decoction pot or an electric ceramic stove.

24. The construction method according to claim 20, characterized in that, The soaking time is 10 to 100 minutes.

25. The construction method according to claim 24, characterized in that, The soaking time is approximately 45 minutes.

26. The construction method according to claim 20, characterized in that, Cover the pot when it boils.

27. The construction method according to claim 20, characterized in that, The power of the container during boiling is 1000-3000W.

28. The construction method according to claim 27, characterized in that, The container has a power of approximately 2000W when boiling.

29. The construction method according to claim 20, characterized in that, The boiling time is 5 to 15 minutes.

30. The construction method according to claim 29, characterized in that, The boiling time is approximately 10 minutes.

31. The construction method according to claim 20, characterized in that, The power of the container during simmering is 500-1500W.

32. The construction method according to claim 31, characterized in that, The power of the container at the simmering stage is approximately 900W.

33. The construction method according to claim 20, characterized in that, The simmering time is 10 to 100 minutes.

34. The construction method according to claim 33, characterized in that, The simmering time is approximately 60 minutes.

35. The construction method according to claim 20, characterized in that, The volume ratio of the decoction containing angelica and cinnamon to the water is 0.1 to 0.

5.

36. The construction method according to claim 35, characterized in that, The volume ratio of the decoction containing angelica and cinnamon to the water is approximately 0.

3.

37. The construction method according to claim 20, characterized in that, The filter screen is a 200-mesh nylon filter screen.

38. The construction method according to claim 1, characterized in that, The concentration of ferulic acid in the reference solution was 10–25 μg / ml.

39. The construction method according to claim 38, characterized in that, The concentration of ferulic acid in the reference solution was approximately 18 μg / ml.

40. The construction method according to claim 1, characterized in that, The concentration of cinnamaldehyde in the reference solution was 10–20 μg / ml.

41. The construction method according to claim 40, characterized in that, The concentration of cinnamaldehyde in the reference solution was approximately 13 μg / ml.

42. The construction method according to claim 1, characterized in that, The concentration of 6-gingerol in the reference solution is 1–10 μg / ml.

43. The construction method according to claim 42, characterized in that, The concentration of 6-gingerol in the reference solution was approximately 6 μg / ml.

44. The construction method according to claim 1, characterized in that, The concentration of ligustilide in the reference solution was 10–25 μg / ml.

45. The construction method according to claim 44, characterized in that, The concentration of ligustilide in the reference solution was approximately 18 μg / ml.

46. ​​The construction method according to claim 1, characterized in that, The reference solution contained 20–40 μg / ml of gallic acid.

47. The construction method according to claim 46, characterized in that, The reference solution contained approximately 30 μg / ml of gallic acid.

48. The construction method according to claim 1, characterized in that, The reference solution contained 50–70 μg / ml of paeoniflorin.

49. The construction method according to claim 48, characterized in that, The reference solution contained approximately 60 μg / ml of paeoniflorin.

50. The construction method according to claim 1, characterized in that, The reference solution contained 40–60 μg / ml of cinnamic acid.

51. The construction method according to claim 50, characterized in that, The reference solution contained approximately 50 μg / ml of cinnamic acid.

52. The construction method according to claim 1, characterized in that, The reference solution contained 40–60 μg / ml of glycyrrhizic acid.

53. The construction method according to claim 52, characterized in that, The reference solution contained approximately 50 μg / ml of glycyrrhizic acid.

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

55. The construction method according to claim 54, characterized in that, The injection volume is approximately 10 μl.

56. The construction method according to claim 1, characterized in that, The theoretical plate number of the chromatographic peak corresponding to the ferulic acid is not less than 5000.

57. The construction method according to claim 1, characterized in that, The chromatographic column is an Agilent 5TC-C18 column.

58. The construction method according to claim 1, characterized in that, When the detection wavelength is 250 nm between 0 minutes and 77 minutes, and 210 nm between 77 minutes and 100 minutes, the fingerprint spectrum includes peaks 1-14, and the retention times of peaks 1-14 are approximately 8.968 min, approximately 21.263 min, approximately 27.481 min, approximately 29.945 min, approximately 38.938 min, approximately 42.241 min, approximately 45.390 min, approximately 64.056 min, approximately 65.959 min, approximately 69.988 min, approximately 72.691 min, approximately 74.593 min, approximately 78.837 min, and approximately 85.261 min, respectively.

59. The construction method according to claim 1, characterized in that, When the detection wavelength is 250 nm between 0 minutes and 77 minutes, and 210 nm between 77 minutes and 100 minutes, 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.

60. The construction method according to claim 1, characterized in that, When the detection wavelength is 250 nm between the 0th and 77th minutes and 210 nm between the 77th and 100th minutes, peak 1 originates from white peony root, peak 2 originates from white peony root, peak 3 originates from white peony root, peak 4 originates from white peony root, peak 5 originates from angelica root, peak 6 originates from stir-fried licorice root, peak 7 originates from white peony root, peak 8 originates from cinnamon root, peak 9 originates from cinnamon root, peak 10 originates from stir-fried licorice root, peak 11 originates from stir-fried licorice root, peak 12 originates from stir-fried licorice root, peak 13 originates from ginger root, and peak 14 originates from angelica root.

61. A method for quality control of a traditional Chinese medicine composition comprising angelica and cinnamon, characterized in that, The quality control method includes the following steps: (1) Establish a standard fingerprint spectrum of a traditional Chinese medicine composition reference sample using the fingerprint spectrum construction method according to any one of claims 1 to 60; (2) Take the test solution of the traditional Chinese medicine composition and detect it under the chromatographic conditions in the fingerprint spectrum construction method according to any one of claims 1 to 60 to obtain the fingerprint spectrum of the traditional Chinese medicine composition to be tested; as well as (3) Compare the fingerprint spectrum of the Chinese medicine composition to be tested obtained in step (2) with the standard fingerprint spectrum of the Chinese medicine composition reference sample obtained in step (1). If the fingerprint spectrum meets the requirements, the product is qualified; if it does not meet the requirements, the product is unqualified.

62. The quality control method according to claim 61, characterized in that, The compliance requirements include one or more of the following: (1) The fingerprint spectrum of the Chinese medicine composition to be tested shows 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 spectrum of the Chinese medicine composition reference sample. (2) Taking peak 4 as peak S, the relative retention time of each characteristic chromatographic peak in the fingerprint spectrum of the Chinese medicine composition sample to be tested and peak S is within ±10% of the relative retention time value of each characteristic chromatographic peak in the standard fingerprint spectrum of the Chinese medicine composition reference sample. as well as (3) According to the similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine, the similarity between the fingerprint of the sample to be tested of the traditional Chinese medicine composition and the standard fingerprint of the reference sample of the traditional Chinese medicine composition shall not be less than 0.

90.

63. Use of the construction method according to any one of claims 1 to 60 or the quality control method according to claim 61 or 62 in the quality detection and / or quality evaluation and / or quality control of traditional Chinese medicine compositions containing Angelica sinensis and cinnamon.

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