Establishment of fingerprint of a mixed plant solid beverage of Poria cocos and cinnamon and determination of its component content

Abstract

The present invention relates to a fingerprint establishment method, a detection method, and a content determination method of the traditional Chinese medicine composition Pinghe Drink. The raw materials of the traditional Chinese medicine composition include: lily, papaya, yam, poria, cinnamon, tangerine peel, polygonatum, polygonatum, and mulberry. The fingerprint establishment method includes the preparation of a reference solution and a test solution; the reference solution and the test solution are detected by high performance liquid chromatography; the mobile phase used in the high performance liquid chromatography is acetonitrile and formic acid aqueous solution, and the elution method is gradient elution. The fingerprint and content determination method of the traditional Chinese medicine composition constructed by the above method can effectively characterize the traditional Chinese medicine composition, and can simply and quickly determine the content of multiple components, providing richer chemical information and a more comprehensive basis for its quality evaluation.

Description

Technical Field

[0001] The invention belongs to the technical field of quality analysis of traditional Chinese medicines, and particularly relates to a high performance liquid chromatography (HPLC) fingerprint establishment and determination method of a tuckahoe, cinnamon mixed plant solid beverage and a component content determination method thereof. Background Art

[0002] Constitution in Traditional Chinese Medicine (TCM) refers to the comprehensive, relatively stable, inherent characteristics of the human body, including its morphological structure, physiological functions, and psychological state, formed through both innate and acquired factors. These characteristics are individual traits that develop during human growth and development, adapting to the natural and social environment. Research on TCM constitutions has found a correlation between constitution and disease, influencing the onset and pattern of disease, as well as the progression of symptoms and prognosis. While constitution is stable, it is also variable. Scientific intervention can facilitate the prevention and cure of disease. Current research categorizes constitutions into nine types based on their overall characteristics and clinical manifestations. Among these, those with a balanced constitution exhibit a balanced balance of yin and yang, qi, and blood, characterized by a moderate physique, a rosy complexion, and abundant energy. This type of constitution is considered relatively healthy and should be prioritized for maintenance.

[0003] The Poria cocos and cinnamon mixed plant solid beverage, marketed as "Pinghe Drink," is a functional food specifically developed by Yabao Jiuhe (Beijing) Health Management Co., Ltd. for people with a balanced constitution. It primarily consists of ingredients such as lily, papaya, yam, Poria cocos, cinnamon, tangerine peel, Polygonatum sibiricum, Polygonatum odoratum, and mulberry. The formula includes lily, which nourishes yin and moistens the lungs, clears the heart and calms the mind; papaya, which relaxes tendons and activates collaterals, harmonizes the stomach and eliminates dampness; yam, which nourishes the spleen and stomach, promotes fluid production and benefits the lungs, and tonifies the kidneys and astringes essence; Poria cocos, which promotes diuresis and eliminates dampness, strengthens the spleen, and calms the mind; cinnamon, which replenishes fire and yang, directs fire back to its source, dispels cold and relieves pain, and warms and unblocks the meridians; tangerine peel, which regulates qi and strengthens the spleen, dries dampness and resolves phlegm; Polygonatum sibiricum, which replenishes qi and nourishes yin, strengthens the spleen, moistens the lungs, and benefits the kidneys; Polygonatum odoratum, which nourishes yin and moistens dryness, promotes fluid production and quenches thirst; and mulberry, which nourishes yin and replenishes blood, promotes fluid production and moistens dryness. Each ingredient in the formula performs its respective functions, making it a suitable dietary therapy for those with a balanced constitution.

[0004] Pinghe Drink (a mixed plant solid beverage containing Poria cocos and cinnamon) is a newly developed product by the applicant. Made from a combination of multiple ingredients, it is complex and lacks quality control methods. Currently, due to the lack of rigorous quality control standards, production is conducted based solely on actual conditions, resulting in varying production processes. This has a negative impact on the standardized production of Pinghe Drink.

[0005] Traditional Chinese medicine compositions contain botanical herbs from diverse sources and with complex compositions. The efficacy of a composition does not derive from a single active ingredient but rather from the combined effects of the ingredients within the composition. Differences in the content and composition ratio of the active ingredients within a composition can affect efficacy. Given the complexity of TCM compositions, any single active ingredient or indicator component is insufficient to effectively evaluate the quality of TCM. The quality of TCM compositions and their preparations should be evaluated using appropriate detection methods that provide rich identification information. Current methods such as microscopic identification, physical and chemical identification, and content determination are insufficient to address this issue.

[0006] At present, the quality control of traditional Chinese medicine compositions such as plant solid beverages mostly uses a single ingredient or component as an evaluation indicator. This has the problem of a single evaluation indicator, which cannot reflect the overall quality level of plant solid beverages, and its quality stability is also difficult to guarantee. Pinghe Drink contains a variety of traditional Chinese medicine ingredients, and the detectable active ingredients may reach dozens or even more. If the different active ingredients in Pinghe Drink are to be identified and determined one by one, a lot of screening work is required. At present, there is no technical report on the establishment of a fingerprint spectrum of Pinghe Drink to detect its quality, which brings more technical obstacles to the establishment of a fingerprint spectrum detection method for Pinghe Drink.

[0007] The fingerprint of traditional Chinese medicine is a comprehensive and quantifiable identification method. It is based on the systematic study of the chemical components of traditional Chinese medicine and is mainly used to evaluate the authenticity, quality and stability of traditional Chinese medicine and semi-finished products of traditional Chinese medicine preparations. It specifically includes two aspects: (1) through the characteristic of the fingerprint, the authenticity of traditional Chinese medicine can be effectively identified; (2) through the main characteristic peak area and ratio of the fingerprint, the quality of traditional Chinese medicine can be effectively controlled. The establishment of the fingerprint of traditional Chinese medicine will be able to more comprehensively reflect the types and quantities of chemical components contained in traditional Chinese medicine and its preparations, and then make an overall description and evaluation of the quality of the medicine. This is also in line with the holistic theory of traditional Chinese medicine. On this basis, if further spectrum efficacy research is carried out, the quality of traditional Chinese medicine can be truly combined with its efficacy, which will help to clarify the mechanism of action of traditional Chinese medicine. The fingerprint of traditional Chinese medicine refers to the chromatogram or spectrum that can indicate the chemical characteristics of traditional Chinese medicine or traditional Chinese medicine preparations after appropriate treatment using certain analytical methods. It is a key technology for achieving the overall correlation quality evaluation of multiple components.

[0008] At present, TCM fingerprint technology has involved many methods, including chromatographic methods such as thin layer scanning (TLCS), high performance liquid chromatography (HPLC), gas chromatography (GC) and high performance capillary electrophoresis (HPCE), as well as spectroscopic methods such as ultraviolet spectroscopy (UV), infrared spectroscopy (IR), mass spectrometry (MS), nuclear magnetic resonance (NMR) and X-ray diffraction.

[0009] Therefore, there is a need for a method for quality monitoring of Pinghe Drink products through fingerprint analysis, as well as a need for a standardized production method based on the quality monitoring method. Summary of the Invention

[0010] In order to improve the above-mentioned technical problems, the present invention provides a method for establishing a fingerprint of a mixed plant solid beverage of Poria cocos and cinnamon and determining the content of its components. The method is accurate, reliable, and simple to operate, providing a basis for quality control of the solid beverage. The mixed plant solid beverage of Poria cocos and cinnamon is a product with complex ingredients. Whether a high-quality fingerprint can be established and accurate content determination can be performed depends on whether the separation effect of the high-performance liquid phase is good. After trial and error, it was verified that the liquid phase conditions provided by the present invention can achieve an excellent separation effect, and multiple components can be separated and peaks can be shown. This is very important for traditional Chinese medicine products with complex ingredients.

[0011] The medicinal ingredients of the Poria cocos and cinnamon mixed plant solid beverage are lily, papaya, yam, Poria cocos, cinnamon, tangerine peel, Polygonatum sibiricum, Polygonatum odoratum and mulberry.

[0012] The fingerprint construction method or detection method of the Poria cocos and cinnamon mixed plant solid beverage of the present invention comprises the following steps of preparing the fingerprint: taking a test solution and performing high performance liquid chromatography detection to obtain a fingerprint. The chromatographic conditions used are: a C18 chromatographic column, 0.1% formic acid aqueous solution as mobile phase A, acetonitrile as mobile phase B, and gradient elution. The gradient elution procedure is:

[0013]

[0014] To make the fingerprint more accurate, the reference solution can be tested under the same liquid phase conditions before and after the test sample is tested, and the chromatographic peak can be identified by the chromatogram of the reference. The reference used in the present invention is hesperidin. The selection of the reference was based on the UPLC-Q-Exactive-Orbitrap-MS technology test results of the inventor's previous Poria cocos and cinnamon mixed plant solid beverage, and then consulted the literature to screen the index components of each herb in the Poria cocos and cinnamon mixed plant solid beverage. At the same time, after experimental testing and other steps, it was finally determined. The reference finally selected had a good peak shape in the chromatogram of the Poria cocos and cinnamon mixed plant solid beverage test solution, without interference from impurity peaks before and after, and the completely separated hesperidin peak was used as the reference peak of the fingerprint.

[0015] The preparation of the reference solution is simple: dissolve the reference substance hesperidin in a solvent capable of dissolving hesperidin. Methanol and / or ethanol are preferred, with methanol being most preferred. There are no specific requirements for the concentration, as long as it meets the upper and lower detection limits of the instrument. The concentrations used in the present invention range from 115 μg / mL to 120 μg / mL, specifically 117.5 μg / mL.

[0016] The test solution is extracted from the Poria cocos and cinnamon mixed plant solid beverage. The extraction process is as follows: the Poria cocos and cinnamon mixed plant solid beverage is added to the solvent for extraction. The solvent is commonly used in the art for extracting natural compounds, including but not limited to alcohols or aqueous alcohol solutions, such as methanol and ethanol. In some embodiments of the present invention, methanol or a 60%-80% aqueous ethanol solution is used, specifically methanol.

[0017] Because the target components of the Poria cocos and cinnamon mixed plant solid beverage are all components with good solubility in alcohol and alcohol water, conventional natural compound extraction methods in the art can be used, such as ultrasonic extraction and reflux extraction. The embodiment of the present invention specifically adopts a water bath reflux process, and the test methanol is heated and refluxed in an 80-degree Celsius water bath for 1 hour. Comparing several different extraction methods, it was found that the chromatogram baseline using methanol water bath reflux extraction was relatively stable, the chromatographic peak was higher, and the peak shape was better, which was the best way.

[0018] After the extraction is completed, the extract is filtered, the filtrate is evaporated to dryness, and the residue is dissolved in a re-dissolution solvent (for example, about 5 ml of methanol), filtered, and the filtrate is taken. The re-dissolution solvent is preferably the same as the extraction solvent. The specific solution concentration after re-dissolution does not need to be forced to be within a certain range, as long as the equipment can detect the components. A more suitable range can be selected: Weight of Poria cocos and cinnamon mixed plant solid beverage: Volume of re-dissolution solvent = (0.5~1.5) g: 1 mL, for example 0.5g, 1g or 1.5g of Poria cocos and cinnamon mixed plant solid beverage / mL.

[0019] The chromatogram obtained under the above conditions has 18 characteristic peaks, among which peak 16 is hesperidin. The fingerprint spectrum uses the chromatographic peak of peak 16 hesperidin as a reference (S). The relative retention times of the 17 common characteristic peaks are:

[0020]

[0021]

[0022] Typically, after a fingerprint spectrum identifies characteristic peaks in a traditional Chinese medicine product, the requirement is satisfied if the peaks are present at their respective locations. However, when a fingerprint spectrum has higher requirements, not only are the peak locations required to meet the requirements, but the areas of the peaks must also meet certain requirements, thereby demonstrating that the proportions and contents of the various components in the composition meet certain requirements.

[0023] When the chromatographic peak of hesperidin No. 16 is used as the reference (S), the relative peak areas of the common characteristic peaks are:

[0024] Peak Relative peak area 1 0.11-0.17 2 0.22～0.33 3 0.07～0.09 4 0.02～0.03 5 0.03～0.04 6 0.02～0.03 7 0.05～0.06 8 0.03～0.04 9 0.05～0.06 10 0.14～0.16 11 0.05～0.06 12 0.09～0.10 13 0.06～0.08 14 0.03～0.04 15 0.36～0.37 16 1 17 0.04～0.05 18 0.03～0.04

[0025] When preparing the fingerprint, in order to prepare the fingerprint more accurately, the chromatographic peaks can be identified by the chromatogram of the reference substance. The fingerprint of the Poria cocos and cinnamon mixed plant solid beverage is prepared using the chromatograms of different batches of the Poria cocos and cinnamon mixed plant solid beverage, preferably at least 5 batches, and preferably at least 10 batches. The fingerprint of the tested Poria cocos and cinnamon mixed plant solid beverage is compared with the fingerprint of the Poria cocos and cinnamon mixed plant solid beverage constructed by the present invention. The relative retention time should be within ±5% of the specified value to be qualified. The product is evaluated for similarity with the fingerprint, and a similarity greater than 0.8, preferably greater than 0.90, is qualified.

[0026] The present invention also provides a method for simultaneously determining the contents of multiple components in the traditional Chinese medicine composition Pinghe Drink (a mixed plant solid beverage of Poria cocos, cinnamon, etc.), comprising the following steps:

[0027] (1) Preparation of reference solution:

[0028] (2) Prepare the test solution:

[0029] (3) HPLC detection of content.

[0030] The present invention adopts the same HPLC conditions as the aforementioned fingerprint detection, and can simultaneously detect the contents of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the Poria cocos and cinnamon mixed plant solid beverage under the same chromatographic conditions. The operation is simple and fast, the reagent consumption is low, and the cost is low. By selecting appropriate detection conditions, the method is able to meet standard requirements in terms of separation, accuracy, repeatability, and stability for determining the contents of each component in the Poria cocos and cinnamon mixed plant solid beverage.

[0031] The reference substance in the reference substance solution is selected from one or more of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin. The reference substance solution can be a solution of each of the four reference substances, or a solution of a mixture of any two or more of the four reference substances.

[0032] HPLC detection and analysis were performed on the reference solution and test solution of each concentration. The test sample concentration was calculated based on the determination results of the reference and test samples, and the content of the test sample components was calculated: a concentration-peak area standard curve of each reference sample component was drawn; the linear regression equation of each component was calculated with the chromatographic peak area Y of the reference solution component of each concentration as the ordinate and the mass concentration X of the reference solution component as the abscissa. The test sample concentration was calculated based on the HPLC peak area of ​​the test sample.

[0033] In one embodiment of the present invention, HPLC detection and analysis are performed on the reference solution and the test solution of each concentration, and the chromatographic peak in the mixed standard (solution of the reference mixture) is qualitatively characterized according to the chromatographic peak of each reference solution. The concentration of the test sample is calculated according to the measurement results of the reference and the test sample, and the content of the test sample component is calculated: a concentration-peak area standard curve of each component in the mixed standard is drawn; the chromatographic peak area Y of the chromatographic peak component of the reference solution of each concentration is used as the ordinate, and the mass concentration X of the reference solution component is used as the abscissa to calculate the linear regression equation of each component, and the test sample concentration is calculated according to the HPLC peak area of ​​the test sample.

[0034] For example, in the content determination process given in the embodiment of the present invention, the linear regression equation is first calculated:

[0035] The linear regression equation for protocatechuic acid was: y = 11480x + 335.7, with a linear range of 2.57 to 12.85 μg / mL; the linear regression equation for chlorogenic acid was: y = 13150x + 292, with a linear range of 1.54 to 7.7 μg / mL; the linear regression equation for ferulic acid was: y = 32176x – 199.5, with a linear range of 1.24 to 6.2 μg / mL; and the linear regression equation for hesperidin was: y = 17998x + 2934.8, with a linear range of 23.5 to 117.5 μg / mL. In these linear regression equations, y represents the peak area of ​​the corresponding component, and x represents the mass concentration of the component in μg / mL. The content of the corresponding substance in the sample was then calculated based on this linear regression equation and the peak area of ​​the sample measured.

[0036] When determining the content in this field, the area comparison method of the test sample and the standard can also be used for calculation, for example, the formula: S 供试品 / S 标准品 =Concentration 供试品 / concentration 标准品 The test sample concentration was calculated in this way.

[0037] For the content determination of the traditional Chinese medicine composition with complex ingredients of the present invention, it is more appropriate to use a linear regression equation to calculate the content of each ingredient.

[0038] The reference substances are dissolved in organic solvents to obtain solutions of the respective reference substances; or multiple reference substances are dissolved in the same organic solvent to obtain solutions of the reference substances. The organic solvent may be selected from methanol and / or ethanol, preferably methanol. When the reference substance solution is a series of concentration reference substance solutions, the concentration ranges of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin are 2-13 μg / mL, 1-8 μg / mL, 1-7 μg / mL, and 20-120 μg / mL, respectively; the concentrations of the reference substances in the series may be increased arithmetic or geometrically.

[0039] In one embodiment of the present invention, the method for preparing the series of reference substance solutions comprises: preparing a solution of a mixture of four reference substances, wherein the concentrations of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the solution are 12.85 μg / mL, 7.7 μg / mL, 6.2 μg / mL, and 117.5 μg / mL, respectively; performing arithmetic dilution on the solution to obtain a series of protocatechuic acid concentrations of 2.57 μg / mL, 5.14 μg / mL, 7.71 μg / mL, 10.28 μg / mL, and 12. 85μg / mL; the concentration series of chlorogenic acid were 1.54μg / mL, 3.08μg / mL, 4.62μg / mL, 6.16μg / mL, 7.7μg / mL; the concentration series of ferulic acid were 1.24μg / mL, 2.48μg / mL, 3.72μg / mL, 4.96μg / mL, 6.2μg / mL; the concentration series of hesperidin were 23.5μg / mL, 47μg / mL, 70.5μg / mL, 94μg / mL, 117.5μg / mL.

[0040] The test solution preparation method is the same as the fingerprint preparation method, but for accurate content calculation, the Poria cocos and cinnamon mixed plant solid beverage granules must be precisely weighed. The ratio of Poria cocos and cinnamon mixed plant solid beverage granules to the post-extraction reconstitution solvent should be controlled in the range of 1.0g:0.5ml-1.5ml, preferably 1.0g:0.9ml-1.1ml.

[0041] The chromatographic column used in the method for preparing fingerprint and content determination of the present invention is a commonly used C18 chromatographic column. Under more optimized conditions, the chromatographic column used is a 250mm×4.6mm, 3-5μm chromatographic column. According to the high performance liquid phase detection of the present invention, the C18 chromatographic column can be AQ-C18 or WELCH AQ-C18 chromatographic column, in one embodiment of the present invention, is WELCH AQ-C18 chromatographic column, for example, C 18 Column (4.6mm×250mm, 5μm). The theoretical plate number should be no less than 5000 according to hesperidin, and the chromatograms of different batches of Poria cocos and cinnamon mixed plant solid beverage and the chromatogram of the reference substance were obtained. AQ-C18 column (4.6 mm × 250 mm, 5 μm) and WELCH AQ-C18 chromatographic column (4.6mm×250mm, 5μm), it was found that The chromatogram obtained using the AQ-C18 column (4.6 mm × 250 mm, 5 μm) showed better peak shapes for each component and the best resolution, making it a preferred column for subsequent studies.

[0042] The peaks of multiple components can basically be realized within the wavelength range of 280-286nm, which can meet the requirements of the preparation of fingerprints. Taking the four reference substances of the present invention as an example, the present invention uses a PDA ultraviolet detector to perform full wavelength scanning on a mixed reference substance containing 4 components. Among them, the reference peak hesperidin has the best absorption effect at 283nm, and protocatechuic acid, chlorogenic acid and ferulic acid have maximum absorption wavelengths at 210nm, 219nm and 207nm, respectively. By comparison, it was found that there was no significant difference between the chromatographic peak response values ​​of protocatechuic acid, chlorogenic acid and ferulic acid at 283nm and the response values ​​at their respective corresponding maximum absorption wavelengths. Taking the above factors into consideration, the present invention selects 283nm or a wavelength near it as the detection wavelength, and the spectral information of each compound is shown in FIG. Figure 3 and Figure 4 .

[0043] There is generally no fixed limit on the injection volume for HPLC; it only needs to ensure ease of use and that the instrument meets the basic detection limit. When peak area accuracy is critical, it is best to maintain the same injection volume for each assay, especially during the entire assay. The injection volume should be consistent throughout the assay, typically 5.0-20.0 μL, for example, 10 μL.

[0044] The flow rate of the mobile phase can be adjusted according to the equipment performance and work efficiency while ensuring good separation of the component peaks. Too high a flow rate will lead to large equipment losses, while too low a flow rate will take a long time. The embodiment of the present invention uses the most commonly used flow rate of 1 ml.min according to the conditions of the instrument and the work efficiency. -1 , and around the flow rate 0.8-1.3ml.min -1 range for detection.

[0045] The present invention does not contain temperature-sensitive substances or operations, so there is no special restriction on the column temperature, and normal detection can be achieved under normal laboratory conditions. Therefore, it is recommended that the HPLC detection environment be 20-40°C, for example, 30°C or 25°C can be set in the embodiments of the present invention.

[0046] The separation effect can be guaranteed within the range of gradient elution conditions given above in the present invention. The most preferred gradient conditions are:

[0047]

[0048]

[0049] In the present invention, "optional", "optionally" or "optionally present" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0050] In the present invention, "comprising" and "including" are open-ended expressions, meaning they include the contents specified in the present invention but do not exclude other aspects. It should be understood that "comprising" and "including" can also include closed-ended expressions, meaning "consisting of."

[0051] Advantageous Effects of the Invention

[0052] (1) By optimizing HPLC chromatographic conditions, the present invention successfully established the fingerprint of the mixed plant solid beverage of Poria cocos and cinnamon, identified a total of 18 common peaks, and simultaneously detected the contents of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the mixed plant solid beverage of Poria cocos and cinnamon, laying the foundation for the establishment of a comprehensive quality standard for the mixed plant solid beverage of Poria cocos and cinnamon.

[0053] (2) The HPLC fingerprint determination method of the mixed plant solid beverage of Poria cocos and cinnamon was tested and proved to be reliable. The quality of the mixed plant solid beverage was comprehensively evaluated. The precision, repeatability and stability tests verified that the method of the present invention has high stability and reliability, and thus can be used as a method for determining the fingerprint of the mixed plant solid beverage of Poria cocos and cinnamon.

[0054] (3) The present invention uses the same chromatographic conditions to simultaneously detect the contents of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the Poria cocos and cinnamon mixed plant solid beverage. The operation is simple and quick, the reagent consumption is low, and the cost is low. By selecting appropriate detection conditions, the method can determine the separation, accuracy, repeatability, and stability of the content of each component in the Poria cocos and cinnamon mixed plant solid beverage and meet the standard requirements. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 A in the middle is the chromatogram of hesperidin reference substance; Figure 1 B in the middle is the fingerprint of the mixed plant solid beverage of Poria cocos, cinnamon and so on; Figure 1 Figure C is an overlay of HPLC fingerprints of 10 batches of Poria cocos and cinnamon mixed plant solid beverages.

[0056] Figure 2 A is the chromatogram of the upper limit of gradient elution; Figure 9 B is the chromatogram of the lower limit of gradient elution.

[0057] Figure 3 A and B are the UV absorption graphs of protocatechuic acid and chlorogenic acid in the wavelength range of 200-400nm, respectively.

[0058] Figure 4 C and D are the ultraviolet absorption graphs of ferulic acid and hesperidin in the wavelength range of 200-400nm, respectively.

[0059] Figure 5 These are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage obtained at different detection wavelengths, where A, B, and C are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage at detection wavelengths of 283nm, 210nm, and 219nm, respectively.

[0060] Figure 6 These are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage obtained under different elution conditions, where A and B are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage under elution conditions one and two, respectively.

[0061] Figure 7 The fingerprints of the mixed plant solid beverage of Poria cocos and cinnamon obtained under different chromatographic columns, where A is Fingerprint of Poria cocos and cinnamon mixed plant solid beverage under AQ-C18 column conditions; B is WELCH Fingerprint of Poria cocos and cinnamon mixed plant solid beverage under AQ-C18 column conditions.

[0062] Figure 8 These are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage obtained under different sample extraction methods, where A, B, and C are the fingerprints of the Poria cocos and cinnamon mixed plant solid beverage under extraction methods A, B, and C, respectively.

[0063] Figure 9 A in the middle is the chromatogram of the test solution of the mixed plant solid beverage of Poria cocos and cinnamon; Figure 9 B is the chromatogram of the mixed reference solution (in the figure: 1-protocatechuic acid; 2-chlorogenic acid; 3-ferulic acid; 4-hesperidin). DETAILED DESCRIPTION

[0064] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0065] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0066] Experimental instruments and reagents:

[0067] 1. Instrument

[0068] High-performance liquid chromatography (LC-20A, Shimadzu Corporation, Japan), magnetic stirring water bath (LC-WB-8+, Shanghai Lichen Bangsi Instrument Technology Co., Ltd.); XS105 electronic balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.); Milli-Q ultrapure water purification system (Beijing Wuzhou Oriental Technology Co., Ltd.).

[0069] 2. Reagents

[0070] Protocatechuic acid (purity ≥98%, batch number 110809-202207, China Food and Drug Administration), chlorogenic acid (purity ≥98%, batch number 110753-202119, China Food and Drug Administration), ferulic acid (purity ≥98%, batch number 110773-202316, China Food and Drug Administration), hesperidin (purity ≥98%, batch number 110721-202220, China Food and Drug Administration) Research Institute); 10 batches (S1-S10) of Poria cocos and cinnamon mixed plant solid beverage, with batch numbers 202204171, 202201221, 202110211, 202104161, 202101311, 2007091, 2005061, 2003141, 2001141, and 1909181, sourced from Shanxi Yabao Jiuhe Pharmaceutical Technology Co., Ltd.; chromatographic grade acetonitrile; chromatographic grade methanol; and ultrapure water.

[0071] Example 1 Establishment of fingerprint of mixed plant solid beverage of Poria cocos and cinnamon

[0072] The fingerprint was prepared according to the following conditions. The relevant results can be found in Figure 1 .

[0073] 1. Chromatographic conditions

[0074] HPLC fingerprint analysis conditions were adopted by WELCH AQ-C18 column (4.6 mm × 250 mm, 5 μm); mobile phase A was water (0.1% formic acid in water), mobile phase B was acetonitrile; gradient elution; flow rate was 1.0 ml min -1 ; Column temperature 30℃; Injection volume 10μL, detection wavelength 283nm.

[0075] Table 1 Gradient elution method

[0076]

[0077]

[0078] 2. Preparation of reference solution

[0079] Take the hesperidin reference substance, accurately weigh it, and add methanol to prepare a hesperidin reference substance solution of 117.50 μg / ml.

[0080] 3. Preparation of test solution

[0081] Take about 5.0 g of the granules of this product, weigh accurately, place them in a stoppered conical flask, add an appropriate amount of methanol, heat and reflux in a water bath at 80 degrees Celsius for 1 hour, cool, filter, evaporate the filtrate to dryness, add 5 ml of methanol to dissolve the residue, filter, and take the filtrate to obtain the product.

[0082] 4. HPLC fingerprint analysis of Poria cocos and cinnamon mixed plant solid beverage

[0083] 4.1 Precision test

[0084] Six consecutive injections of a mixed plant solid beverage containing Poria cocos and cinnamon from the same batch (Batch No. 202204171) were performed. The retention time and peak area of ​​each common chromatographic peak were recorded. The relative retention time and peak area of ​​hesperidin, peak 16, were used as a reference to calculate the relative retention time and peak area of ​​each common peak. This was done to examine the consistency of the relative retention time and peak area ratios of the chromatographic peaks. The results showed that the relative retention time RSDs for each common peak were less than 1.36%, and the relative peak area RSDs were less than 3.82%, demonstrating that the assay method of the present invention is stable, has good system integrity, and is reliable.

[0085] 4.2 Stability test

[0086] Take the same batch (batch number: 202204171) of Poria cocos and cinnamon mixed plant solid beverage test sample, at different time points (0h, 2h, 4h, 6h, 8h, 12h, 24h) sample analysis, record the retention time and peak area of ​​each common chromatographic peak, with the retention time and peak area of ​​peak 16 hesperidin as a reference, convert the relative retention time and relative peak area of ​​each common peak to investigate the consistency of the relative retention time and relative peak area ratio of the chromatographic peak. The results show that the relative retention time RSD of each common peak is less than 1%, and the relative peak area RSD is less than 3.15%. This shows that the Poria cocos and cinnamon mixed plant solid beverage sample of the present invention has good stability under experimental conditions and can ensure the reliability of the analysis results within 24h.

[0087] 4.3 Repeatability test

[0088] Take the same batch (batch number: 202204171) of Poria cocos and cinnamon mixed plant solid beverage test sample, and prepare 6 test sample solutions in parallel according to the method under "3". Samples were injected and analyzed separately, and the retention time and peak area of ​​each common chromatographic peak were recorded. The retention time and peak area of ​​peak 16 hesperidin were used as a reference to convert the relative retention time and relative peak area of ​​each common peak to examine the consistency of the relative retention time and relative peak area ratio of the chromatographic peaks. It was shown that the relative retention time RSD of each common peak was less than 1%, and the relative peak area RSD was less than 3.76%, indicating that the method of the present invention is stable and reliable and conducive to promotion.

[0089] 4.4 Establishment and similarity evaluation of HPLC fingerprint of Poria cocos and cinnamon mixed plant solid beverage

[0090] Ten batches of a mixed plant beverage containing Poria cocos and cinnamon (batch numbers: 202204171, 202201221, 202110211, 202104161, 202101311, 2007091, 2005061, 2003141, 2001141, and 1909181) were prepared and tested according to the method in item 3. Samples were injected and analyzed, and chromatograms were recorded. A representative chromatogram (batch number 202204171 of a mixed plant beverage containing Poria cocos and cinnamon) was selected as the reference fingerprint. The "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines" recommended by the Pharmacopoeia Commission was used for analysis. Chromatographic similarity was used as the fingerprint evaluation metric, and multi-point calibration and full-spectrum matching were used to generate the common fingerprint pattern. The similarities between the fingerprints of each batch of samples and the control were (0.999, 0.999, 0.997, 0.997, 0.996, 0.996, 0.996, 0.995, 0.996, 0.995), respectively.

[0091] 4.5 Identification of common peaks in fingerprints, calculation of relative peak areas and relative retention times

[0092] Using similarity evaluation system software, a total of 18 peaks were detected in the 10 fingerprints. Comparison with the reference sample identified peak 16 as hesperidin, with a retention time of 84.700 min. Peak 16 had a good peak shape, with no interference from impurities before and after, and was completely separated. Therefore, it was selected as a reference peak for calculating the relative peak areas and relative retention times of the other shared peaks. The results are shown in Tables 2 and 3 below.

[0093] Table 2 Relative peak areas of 18 common peaks in 10 batches of samples (S1-S10)

[0094]

[0095] Table 3 Relative retention times of 18 common peaks in 10 batches of samples (S1-S10)

[0096]

[0097]

[0098] 4.6 Range of Gradient Elution Conditions

[0099] By adjusting the conditions of gradient elution, it can be found that under the currently determined elution conditions, the ratio of mobile phases A and B can be appropriately adjusted, and the upper and lower limits of the elution conditions can be determined through experiments.

[0100] Upper elution condition:

[0101]

[0102] Lower limit elution conditions:

[0103]

[0104]

[0105] Results see Figure 2 , Figure 2 A is the upper limit elution condition of the mobile phase, Figure 2 Figure B is the lower limit elution condition of the mobile phase. Both conditions meet the effect of good separation and complete elution.

[0106] Example 2 Comparison of Fingerprint Preparation Methods of Poria Cocos and Cinnamon Mixed Plant Solid Beverage

[0107] 1. Determination of chromatographic conditions and exploration of fingerprint methods

[0108] The fingerprint results of Pinghe Drink prepared under different HPLC chromatographic conditions were investigated and screened.

[0109] (1) Detection wavelength

[0110] Prepare and test according to the method of Example 1. Take about 5.0g of the granules of this product, weigh accurately, place in a stoppered conical flask, add an appropriate amount of methanol, heat under reflux in a water bath at 80 degrees Celsius for 1 hour, cool, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of methanol, filter, and take the filtrate.

[0111] HPLC fingerprint analysis conditions were adopted by WELCH AQ-C18 column (4.6 mm × 250 mm, 5 μm); mobile phase A was water (0.1% formic acid in water), mobile phase B was acetonitrile; gradient elution; flow rate was 1.0 ml min -1; column temperature 30 ℃; injection volume 10 μL, analysis at detection wavelengths of 283 nm, 210 nm and 219 nm, respectively. The results are shown in Figure 5 .

[0112] As can be seen, using 283nm as the detection wavelength provides richer chromatographic information, a more stable baseline, and more balanced responses across peaks, which comprehensively reflects the components in the sample. Therefore, 280-286nm was selected as the detection wavelength for the fingerprint of the Poria cocos and cinnamon mixed plant solid beverage sample, with the optimal condition being 283nm.

[0113] (2) Elution conditions

[0114] The inventors compared several different elution conditions and the results showed that the mobile phase elution conditions currently selected by the present invention are the best conditions. Due to the large number of elution conditions, the following is a brief example: Take the particles of this product, prepare them according to the method for preparing the test solution in Example 1, and compare the following two elution conditions according to the chromatographic conditions in (1) detection wavelength. The results are shown in FIG. Figure 6 As can be seen, in the chromatogram under elution condition 1, the peak shapes of the components are good and the separation is good. Therefore, elution condition 1 was selected as the elution condition for the fingerprint of the Poria cocos and cinnamon mixed plant solid beverage sample.

[0115] Elution condition 1:

[0116]

[0117] Elution condition 2:

[0118]

[0119] (3) Column type

[0120] Take the granules of this product and use the chromatographic columns according to the preparation method and chromatographic conditions of the test solution in Example 1. AQ-C18 (4.6 mm × 250 mm, 5 μm) and WELCH AQ-C18 (4.6mm×250mm, 5μm) was used for analysis. The results are shown in Figure 7 It can be seen that both can meet the test of fingerprint spectrum and can be used as chromatographic columns for preparing fingerprint spectrum. The peak shape of each component of the chromatogram separated by AQ-C18 column (4.6mm×250mm, 5μm) is good, and the separation is good. An AQ-C18 chromatographic column (4.6 mm × 250 mm, 5 μm) was used as the chromatographic column for the fingerprint of the Poria cocos and cinnamon mixed plant solid beverage test sample.

[0121] 3. Determination of sample extraction method

[0122] Pingheyin has a complex composition, and different extraction solvents can affect the extraction of the active ingredients. Finding the most suitable extraction method is crucial to maximize the extraction of ingredients and to visualize the peaks on the HPLC graph, which is one of the technical difficulties.

[0123] Samples were extracted using methods A, B, and C. The processes are as follows:

[0124] A: Take about 5.0g of this product granules, accurately weighed, placed in a stoppered conical flask, add appropriate amount of methanol, heat under reflux in a water bath at 80 degrees Celsius for 1 hour, cool, filter, evaporate the filtrate to dryness, add 5ml of methanol to dissolve the residue, filter, and take the filtrate to obtain.

[0125] B: Take about 5.0g of the granules of this product, weigh accurately, place in a stoppered conical flask, add an appropriate amount of methanol, ultrasonically treat for 1h, cool, filter, evaporate the filtrate to dryness, add 5ml of methanol to dissolve the residue, filter, and take the filtrate to obtain.

[0126] C: Take about 5.0g of the granules of this product, weigh accurately, place in a stoppered conical flask, add an appropriate amount of 70% ethanol, heat under reflux in a water bath at 80 degrees Celsius for 1 hour, cool, filter, evaporate the filtrate to dryness, add 5ml of 70% ethanol to dissolve the residue, filter, and take the filtrate to obtain.

[0127] The test solutions prepared by the above three methods were tested according to the chromatographic conditions finally determined above. The results are as follows: Figure 8 As shown, from Figure 8 All three methods meet the requirements for fingerprint preparation and can be used as extraction methods for fingerprint preparation or content determination. However, after comparison, methanol water bath reflux extraction was found to be the best, producing the most stable chromatogram baseline, the best chromatographic peak separation, and the best peak shape.

[0128] Example 3 Method for Determining the Contents of Four Components in Poria Cocos and Cinnamon Mixed Plant Solid Beverage

[0129] 1. Chromatographic conditions

[0130] Same as Example 1.

[0131] 2. Preparation of reference solution

[0132] Take appropriate amounts of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin reference substances, accurately weigh them, and add methanol to prepare a mixed reference solution containing 12.85 μg / mL of protocatechuic acid, 7.70 μg / mL of chlorogenic acid, 6.20 μg / mL of ferulic acid, and 117.50 μg / mL of hesperidin.

[0133] At the same time, appropriate amounts of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin reference substances were taken and accurately weighed, and methanol was added to prepare single solutions of each reference substance at 12.85 μg / mL of protocatechuic acid, 7.70 μg / mL of chlorogenic acid, 6.20 μg / mL of ferulic acid, and 117.50 μg / mL of hesperidin.

[0134] 3. Preparation of test solution

[0135] Take about 5.0 g of the granules of this product, weigh accurately, place them in a stoppered conical flask, add an appropriate amount of methanol, heat and reflux in a water bath at 80 degrees Celsius for 1 hour, cool, filter, evaporate the filtrate to dryness, add 5 ml of methanol to dissolve the residue, filter, and take the filtrate to obtain the product.

[0136] 4. Content determination and methodological investigation of Poria cocos and cinnamon mixed plant solid beverage

[0137] 4.1 Precision test

[0138] Six consecutive injections of the same batch number (202204171) of a mixed plant solid beverage containing Poria cocos and cinnamon were performed, and the peak areas were recorded. The RSD values ​​for the peak areas of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin were calculated to be 0.63%, 1.7%, 0.68%, and 0.77%, respectively, demonstrating the good precision of the assay method.

[0139] 4.2 Linear relationship investigation

[0140] According to the chromatographic conditions under item "1" above, take a single solution of each reference substance for chromatographic detection, and then perform chromatographic detection on the solution of the reference substance mixture. Compare the chromatographic peaks of the single reference substance to confirm the ownership of the chromatographic peaks of the reference substance mixture. Then, directly perform chromatographic analysis on the solution of the reference substance mixture to simultaneously determine the four components and obtain a linear equation.

[0141] Accurately measure 1 mL, 2 mL, 3 mL, 4 mL, and 5 mL of the mixed reference solution into 5 mL volumetric flasks. Dilute to the mark with methanol to prepare mixed reference solutions of varying concentrations. Analyze according to the chromatographic conditions in "1" above, and record the liquid chromatograms. Plot a standard curve using the peak area (Y) of the chromatographic peak of each mixed reference solution concentration as the ordinate and the concentration (X, μg / mL) of each mixed reference solution as the abscissa. Calculate the regression equation and correlation coefficient for each component (see Table 4).

[0142] Table 4 Regression equations, correlation coefficients, and linear ranges of the four chemical components

[0143]

[0144] 4.3 Stability test

[0145] The same batch number (202204171) of Poria cocos and cinnamon mixed plant solid beverage was taken as the test sample. According to the chromatographic conditions under item "1", the test solution was sampled and analyzed after 0h, 2h, 4h, 6h, 8h, 10h, 12h, and 24h, and the peak areas were recorded respectively. The RSD values ​​of the peak areas of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin were calculated to be 1.12%, 0.58%, 0.79%, and 0.76%, respectively, indicating that the test solution of the present invention is stable within 24h.

[0146] 4.4 Repeatability test

[0147] Take the same batch number (202204171) of Poria cocos and cinnamon mixed plant solid beverage test sample, and prepare 6 test sample solutions in parallel according to the method under item "3". The samples were injected and analyzed according to the chromatographic conditions under item "1", and the peak areas were recorded respectively. The linear regression equations of each component calculated the contents of each component, which were: protocatechuic acid 11.99 μg / mL, chlorogenic acid 7.38 μg / mL, ferulic acid 5.87 μg / mL, and hesperidin 106.47 μg / mL. The peak area RSD values ​​were 1.23%, 0.93%, 1.26%, and 0.76%, respectively. This shows that the method of the present invention has good repeatability.

[0148] 4.5 Sample recovery

[0149] Take a known amount of Poria cocos and cinnamon mixed plant solid beverage (202204171), pour out the contents, accurately weigh 2.50g, and prepare 6 parallel portions. Add 1mL of a mixed reference solution containing 30μg / mL protocatechuic acid, 20μg / mL chlorogenic acid, 15μg / mL ferulic acid, and 265μg / mL hesperidin. Prepare the test solution according to the method under "3" and inject and analyze according to the chromatographic conditions under "1". Record the peak area and calculate the recovery rate of each component by the linear regression equation. The results are shown in Table 5. The calculation method for "sample content" in Table 5 is: multiply the content of each component measured in 4.4 by 5ml and divide by 2.

[0150] Table 5 Recovery results of the four components of the mixed plant solid beverage of Poria cocos and cinnamon (n=6)

[0151]

[0152]

[0153] 4.6 Sample determination

[0154] Ten batches of the test solution of the Poria cocos and cinnamon mixed plant solid beverage were taken and sampled and analyzed according to the chromatographic conditions under item "1". The peak area of ​​each component was recorded respectively, and the content of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin was calculated respectively by the linear regression equation of each component. The results are shown in Table 6 below.

[0155] Table 6 Determination results of the contents of four components in 10 batches of Poria cocos and cinnamon mixed plant solid beverage (μg / mL, n=3)

[0156]

[0157] In summary, the present invention successfully established a fingerprint of the Poria cocos and cinnamon mixed plant solid beverage by optimizing the HPLC chromatographic conditions, identified a total of 18 common peaks, and was able to simultaneously detect the contents of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the Poria cocos and cinnamon mixed plant solid beverage. Methodological verification showed that all indicators of the detection method of the present invention were good, thereby improving the quality control standard of the Poria cocos and cinnamon mixed plant solid beverage and providing a reference basis for the establishment of its comprehensive quality standard.

[0158] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A method for constructing or detecting a fingerprint of a Chinese medicine composition, Poria cocos and cinnamon mixed plant solid beverage, wherein the raw materials of the Chinese medicine composition are composed of: lily, papaya, yam, Poria cocos, cinnamon, tangerine peel, polygonatum, polygonatum, and mulberry. A test solution is subjected to high performance liquid chromatography to obtain a fingerprint; The chromatographic conditions are as follows: the chromatographic column is a C18 column, 0.1% formic acid aqueous solution is used as mobile phase A, acetonitrile is used as mobile phase B, and gradient elution is performed. The gradient elution program is: The detection wavelength of the high performance liquid phase is 280-286 nm; The preparation method of the test solution comprises the following steps: taking a mixed plant solid beverage of Poria cocos and cinnamon, adding an extraction solvent for extraction, filtering the extract after extraction, evaporating the filtrate to dryness, adding an extraction solvent for redissolution, filtering, and taking the filtrate as the test solution; the extraction solvent is alcohol or an aqueous solution of alcohol, and the alcohol is selected from methanol and ethanol; and the extraction process is water bath reflux or ultrasonic extraction.

2. The method according to claim 1, further comprising a high performance liquid phase detection step of a reference substance solution, wherein the reference substance is hesperidin.

3. The method according to claim 2, wherein the reference substance solution is prepared as follows: take the reference substance hesperidin and dissolve it in methanol or ethanol.

4. The method according to claim 1, wherein the fingerprint has 18 characteristic peaks, wherein peak 16 is hesperidin, and the chromatographic peak of peak 16 of hesperidin is used as the reference peak in the fingerprint. The relative retention times of the 17 common characteristic peaks are: 。 5. The method according to claim 1, wherein when the chromatographic peak of hesperidin No. 16 is used as the reference peak in the fingerprint, the relative peak areas of the common characteristic peaks are: 。 6. A method for simultaneously determining the contents of multiple components in a Chinese medicine composition, Poria cocos and cinnamon mixed plant solid beverage, wherein the raw materials of the Chinese medicine composition are composed of: lily, papaya, yam, Poria cocos, cinnamon, tangerine peel, polygonatum, polygonatum, and mulberry, characterized in that: The reference solution and the test solution were analyzed by HPLC. The concentration of the test sample was calculated based on the determination results of the reference solution and the test sample, and the content of the test sample components was calculated. The chromatographic conditions are as follows: the chromatographic column is a C18 column, 0.1% formic acid aqueous solution is used as mobile phase A, acetonitrile is used as mobile phase B, and gradient elution is performed. The gradient elution program is: The detection wavelength is 280-286 nm. The reference substance is selected from one or more of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin, and the reference substance solution is a solution of each of the four reference substances, or a mixed solution of any two or more reference substances; The preparation method of the test solution comprises: extracting a mixed plant solid beverage of Poria cocos and cinnamon, adding an extraction solvent, filtering the extract after extraction, evaporating the filtrate to dryness, re-dissolving it with the extraction solvent, filtering, and taking the filtrate as the test solution; when performing content determination, the ratio of the mixed plant solid beverage particles of Poria cocos and cinnamon to the re-dissolving solvent after extraction is 1.0 g: 0.9 ml-1.1 ml; the extraction solvent is an alcohol or an aqueous solution of an alcohol, and the alcohol is selected from methanol and ethanol; and the extraction process is water bath reflux or ultrasonic extraction.

7. The method according to claim 6, wherein the reference substance solution is prepared by: dissolving each reference substance in an organic solvent to obtain a solution of each reference substance; or dissolving multiple reference substances in the same organic solvent to obtain a solution of a reference substance mixture, wherein the organic solvent is selected from methanol or ethanol.

8. The method according to claim 7, wherein the concentration ranges of protocatechuic acid, chlorogenic acid, ferulic acid, and hesperidin in the reference solution are 2-13 μg / mL, 1-8 μg / mL, 1-7 μg / mL, and 20-120 μg / mL, respectively; and the concentration series of each reference substance increases in arithmetic or geometric increments. The method for calculating the content of the test sample is the linear regression equation method: draw a concentration-peak area standard curve for each reference sample component; use the chromatographic peak area Y of the reference sample solution component of each concentration as the vertical coordinate and the mass concentration X of each reference sample solution component as the horizontal coordinate to calculate the linear regression equation of each component; according to the HPLC peak area of ​​the test sample, substitute it into the linear regression equation to calculate the test sample concentration.

9. The method according to claim 1 or 6, wherein the C18 chromatographic column is a 250 mm×4.6 mm, 3-5 μm chromatographic column.

10. The method according to claim 9, wherein the C18 chromatographic column is a 5 μm chromatographic column. The method according to claim 1 or 6 , wherein the detection wavelength is 283 nm.

12. The method according to claim 1 or 6, wherein the chromatographic conditions are: Chromatographic column: C18 column, 250 mm × 4.6 mm, 5 μm, detection wavelength 283 nm; The gradient elution program was: 。 13. The method according to claim 1 or 6, wherein the extraction process comprises adding methanol and heating under reflux in a water bath at 80 degrees Celsius for 0.5-2 hours, filtering the extract, evaporating the filtrate to dryness, adding 5 ml of methanol to dissolve the residue, filtering, and taking the filtrate to obtain the product.

14. Use of the method according to any one of claims 1 to 13 in the quality control of a Poria cocos and cinnamon mixed plant solid beverage; comparing the fingerprint of the tested Poria cocos and cinnamon mixed plant solid beverage with the fingerprint of a standard Poria cocos and cinnamon mixed plant solid beverage; a product is qualified if the relative retention time is within ±5% of the specified value; and a similarity evaluation is performed with the standard fingerprint; a product is qualified if the similarity is greater than 0.8.

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