Heart nourishing and palpitation calming capsule quality control method based on HPLC fingerprint spectrum

Abstract

The invention provides a quality control method for heart-nourishing and palpitation-calming capsules based on an HPLC (High Performance Liquid Chromatography) fingerprint spectrum, and belongs to the technical field of medicine detection. The method comprises the following steps: weighing capsule contents to prepare a test solution, setting liquid phase conditions, carrying out methodological verification, constructing a fingerprint spectrum, and importing the fingerprint spectrum into software to carry out similarity analysis. And further performing clustering analysis and principal component analysis on the common peak area by adopting SPSS software so as to evaluate the quality consistency among batches. Results show that the consistency of the fingerprints of all batches of samples is good, the similarity reaches 0.991 or above, and the uniformity of the sample quality is further verified through clustering analysis and principal component analysis. The method provided by the invention is comprehensive and accurate, provides a scientific basis for quality control of the heart-nourishing and palpitation-calming capsules, and ensures the stability and consistency of medicine quality.

Description

Technical Field

[0001] The invention belongs to the technical field of drug detection, and particularly relates to a quality control method for Yangxin Dingji capsules based on HPLC fingerprint. Technical Background

[0002] Yangxin Dingji Capsules, a traditional Chinese medicine, are commonly used clinically to treat symptoms such as palpitations, shortness of breath, and fatigue. They nourish the blood and qi, strengthen the pulse, and calm palpitations, improving heart function and alleviating related symptoms. Their main ingredients are a variety of Chinese medicinal herbs, prepared using a specific process.

[0003] In the process of drug production and quality control, ensuring the stability and consistency of drug quality is the key to ensuring the safety and effectiveness of drug use for patients. At present, there are certain limitations in the traditional methods for the quality control of Yangxin Dingji Capsules. Some methods only focus on the content determination of certain single ingredients, while ignoring the synergistic effects of other ingredients in the drug on the efficacy. At the same time, some detection methods are relatively cumbersome in the operation process and have high requirements for experimental conditions. Large errors may occur when different experimenters operate, making it difficult for the test results to accurately reflect the true quality status of the drug. In addition, with the continuous improvement of drug quality standards, traditional quality control methods have been unable to meet the requirements of the current "Pharmacopoeia of the People's Republic of China" and related regulations for comprehensive and precise control of drug quality.

[0004] Therefore, developing a quality control method for Yangxin Dingji Capsules based on HPLC fingerprint to comprehensively and accurately evaluate the quality of drugs and ensure the consistency and stability of the quality of different batches of drugs has become an urgent problem to be solved. Summary of the Invention

[0005] The object of the present invention is to provide a quality control method for Yangxin Dingji Capsules based on HPLC fingerprint. The method establishes the quality control standard of Yangxin Dingji Capsules through HPLC fingerprint technology, and provides a scientific basis for the quality evaluation and quality control of the drug by analyzing the quality consistency of samples from different batches.

[0006] The object of the present invention is achieved through the following technical solutions:

[0007] The present invention provides a quality control method for Yangxin Dingji Capsules based on HPLC fingerprint, comprising the following steps:

[0008] (1) Preparation of test solution: Weigh the contents of Yangxin Dingji Capsules, add methanol, sonicate, make up to the loss, and filter to prepare the test solution;

[0009] (2) Liquid phase conditions: Acetonitrile was used as mobile phase A, 0.1% phosphoric acid aqueous solution was used as mobile phase B, and an UltimatePlus C18 column was used for liquid phase conditions.

[0010] (3) Methodological validation: Verify the reliability of the method through precision tests, stability tests, and repeatability tests;

[0011] (4) Fingerprint construction: multiple batches of sample solutions were injected into the liquid phase system to generate fingerprints and then imported into the Chinese medicine chromatographic fingerprint similarity evaluation software for similarity analysis;

[0012] (5) Multivariate statistical analysis: SPSS software was used to perform cluster analysis and principal component analysis on the common peak areas to evaluate the quality consistency between batches.

[0013] Furthermore, in step (1), the specific preparation method of the test solution is as follows: weigh 0.5 g of the content of Yangxin Dingji Capsules and place it in a conical flask, add 25 ml of 50% methanol solution, accurately add 25 mL of 50% methanol solution, weigh, ultrasonically treat, cool, weigh again, make up the lost amount with 50% methanol solution, shake well, filter, and filter the filtrate with a 0.45 μm microporous filter membrane to obtain.

[0014] Furthermore, the ultrasonic treatment is carried out for 30 minutes at a power of 100 W and a frequency of 40 kHz.

[0015] Furthermore, in step (2), the liquid phase conditions include a flow rate of 1.0 mL min -1 ; Detection wavelength 265nm; injection volume 10μL; column temperature 30℃.

[0016] Furthermore, in step (2), the gradient elution procedure is as follows:

[0017]

[0018] Furthermore, in step (3), the precision test uses chromatographic peak No. 7 as the reference peak to calculate the RSD values ​​of the relative retention time and peak area of ​​each common peak, requiring RSD < 2%;

[0019] The stability test was completed within 24 hours at room temperature, and the chromatographic peak No. 7 was used as the reference peak to calculate the relative retention time and peak area RSD values ​​of the common peaks, with the requirement that RSD < 2%;

[0020] The repeatability test was performed by preparing 6 parallel test solutions, taking the chromatographic peak No. 7 as the reference peak, and calculating the RSD values ​​of the relative retention time and peak area of ​​each common peak, with the RSD requirement being <2%.

[0021] Furthermore, in step (5), the cluster analysis adopts the inter-group linkage method and uses the squared Euclidean distance as the classification basis to divide the sample quality.

[0022] Furthermore, in step (5), the principal component analysis selects the principal components based on the condition that the eigenvalue is greater than 1.

[0023] The beneficial effects of the present invention are:

[0024] The present invention uses HPLC fingerprint technology to comprehensively and systematically analyze the chemical composition information in Yangxin Dingji Capsules. Compared to single-component detection, this technology can simultaneously obtain characteristic information of multiple components, thereby constructing a representative fingerprint spectrum, establishing a more scientific and accurate quality control standard for Yangxin Dingji Capsules. This quality control method based on the overall chemical composition characteristics can better reflect the true quality status of the drug, effectively avoiding the one-sidedness that may be caused by single-component detection, and providing a solid guarantee for the stability and controllability of drug quality.

[0025] Using the method presented herein, different batches of Yangxin Dingji Capsules were analyzed, visually demonstrating the consistency of the fingerprints across batches. The experimental results showed good consistency across the fingerprints of each batch, with similarities to the referenced fingerprint reaching 0.991 or higher. This demonstrates the consistent and stable quality of each batch of Yangxin Dingji Capsules. This result not only provides an important basis for quality control during drug production but also helps ensure the efficacy and safety of different batches of drugs in clinical use, enabling patients to receive stable and reliable treatment.

[0026] Through a series of methodological investigations such as precision tests, stability tests, and repeatability tests, the reliability, stability, and repeatability of the method adopted in the present invention were fully verified. The results of the precision test showed that the RSD values ​​of the relative retention time and relative peak area were both less than 2%, indicating that the method had good consistency in different measurements; the stability test showed that the test sample was stable within 24 hours, ensuring the quality stability of the sample during the analysis process; the repeatability test further confirmed the reliability of the method under multiple repeated operations. These methodological investigation results provide solid theoretical support for the practical application of this quality control method and ensure the accuracy and credibility of the analysis results. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0028] Figure 1 The fingerprints of 10 batches of Yangxin Dingji Capsules in Example 1 of the present invention are as follows;

[0029] Figure 2 This is a cluster analysis diagram of 10 batches of Yangxin Dingji Capsule samples in Example 1 of the present invention;

[0030] Figure 3 This is the principal component analysis scree plot in Example 1 of the present invention;

[0031] Figure 4 This is a chromatogram of the precision test in Example 1 of the present invention;

[0032] Figure 5 This is a chromatogram of the stability test in Example 1 of the present invention;

[0033] Figure 6 It is the repeatability test chromatogram in Example 1 of the present invention. DETAILED DESCRIPTION

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0036] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0037] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.

[0038] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0039] Example 1

[0040] This example aims to establish the quality control standard of Yangxin Dingji Capsule by HPLC fingerprint technology, analyze the quality consistency of different batches of samples, and provide a scientific basis for the quality evaluation and quality control of the drug. The specific research methods and results are as follows:

[0041] 1. Instruments and Materials

[0042] Shimadzu (Shimadzu Enterprise Management (China) Co., Ltd.), JJ324BC balance (Changshu Shuangjie Testing Instrument Factory), SB25-12DT CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0043] Yangxin Dingji Capsules (Hebei Yongfeng Pharmaceutical Co., Ltd., batch numbers 06724110141, 06724110241, 06724110341, 06724110441, 06724110541, 06724110641, 06724120241, 06724030141, 06724030241, 06724030341), acetonitrile (batch number 02250120) and methanol (batch number 0201801756) were all chromatographically pure, and phosphoric acid (batch number 02045202) was analytically pure and were purchased from Tianjin Komiou Chemical Reagent Co., Ltd.

[0044] 2. Methods

[0045] 2.1 Preparation of test solution

[0046] Weigh 0.5 g of the contents of Yangxin Dingji Capsules and place it in a conical flask, add 25 ml of 50% methanol solution, accurately add 25 mL of 50% methanol solution, weigh, treat with ultrasound (power 100 W, frequency 40 kHz) for 30 min, let cool, weigh again, make up the lost amount with 50% methanol solution, shake well, filter, and filter the filtrate with a 0.45 μm microporous filter membrane to obtain the product.

[0047] 2.2 Liquid phase conditions

[0048] Acetonitrile was used as mobile phase A, and 0.1% phosphoric acid aqueous solution was used as mobile phase B. The chromatographic column was an Ultimate Plus C18 column (4.6×250 nm, 5 μm). The flow rate was 1.0 mL·min -1 ; Detection wavelength 265nm; injection volume 10μL; column temperature 30℃; specific gradient elution program is shown in Table 1.

[0049] Table 1 Gradient elution program

[0050]

[0051] 2.3 Methodological Investigation

[0052] Precision test: Prepare the test solution according to the method in "2.1" and make 6 consecutive injections under the chromatographic conditions in "2.2". The chromatogram is shown in Figure 4 , record the peak area, and calculate the relative retention time and relative peak area of ​​each common peak using chromatographic peak No. 7 as a reference. The results show that the RSD values ​​of relative retention time and relative peak area are both less than 2%, indicating good precision of the method.

[0053] Stability test: Prepare the test solution according to the method in "2.1". Place it at room temperature for 0, 2, 4, 8, 12, and 24 hours. Then inject the sample and measure it under the chromatographic conditions in "2.2". The chromatogram is shown in Figure 5 , record the peak area, and calculate the relative retention time and relative peak area of ​​each common peak using chromatographic peak No. 7 as a reference. The results show that the RSD values ​​of relative retention time and relative peak area are both less than 2%, indicating that the test sample has good stability within 24 hours.

[0054] Repeatability test: Prepare 6 portions of the test solution according to the method in "2.1" and inject the sample for determination under the chromatographic conditions in "2.2". The chromatogram is shown in Figure 6 , record the peak area, and calculate the relative retention time and relative peak area of ​​each common peak using chromatographic peak No. 7 as a reference. The results show that the RSD values ​​of relative retention time and relative peak area are both less than 2%, indicating that the method has good reproducibility.

[0055] 2.4 Fingerprint establishment and similarity analysis

[0056] The sample solutions of 10 batches of Yangxin Dingji Capsules were prepared according to the above method and injected under the set chromatographic conditions. The data were imported into the Chinese medicine chromatographic fingerprint similarity evaluation software (2012 version) to obtain the fingerprints of 10 batches of Yangxin Dingji Capsules, as shown in Figure 2. Figure 1 As shown, the fingerprints of each batch of samples showed good consistency, indicating that this method can effectively reflect the chemical composition characteristics of Yangxin Dingji Capsules. Similarity analysis of 10 batches of samples using the software showed that the similarity between all 10 batches of samples and the reference spectrum reached 0.991 or above, indicating a high degree of similarity, indicating that the quality of Yangxin Dingji Capsules is relatively consistent and stable across batches.

[0057] 2.5 Cluster Analysis

[0058] The 15 common peak areas of 10 batches of Yangxin Dingji Capsule samples were imported into SPSS software and the samples were classified using the intergroup linkage method. The results are as follows: Figure 2 When the squared Euclidean distance is 5, the 10 batches of samples can be divided into two categories: S2, S4, S8, and S10 are in one category, and S1, S3, S5-S7, and S9 are in another category. The sample quality is relatively uniform.

[0059] 2.6 Principal Component Analysis

[0060] The 15 common peak areas of 10 batches of Yangxin Dingji Capsule samples were imported into SPSS software. Based on the screening condition of eigenvalue greater than 1, 6 principal components were successfully identified. The cumulative variance contribution rate of these principal components was as high as 92.464%, which fully reflected the main characteristics of the original data. Figure 3 shown.

[0061] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A quality control method for Yangxin Dingji Capsules based on HPLC fingerprint, characterized in that: The following steps are involved: (1) Preparation of test solution: Weigh the contents of Yangxin Dingji Capsules, add methanol, sonicate, make up to the loss, and filter to prepare the test solution; (2) Liquid phase conditions: Acetonitrile was used as mobile phase A, 0.1% phosphoric acid aqueous solution was used as mobile phase B, and an UltimatePlus C18 column was used for liquid phase conditions. (3) Methodological validation: Verify the reliability of the method through precision tests, stability tests, and repeatability tests; (4) Fingerprint construction: multiple batches of sample solutions were injected into the liquid phase system to generate fingerprints and then imported into the Chinese medicine chromatographic fingerprint similarity evaluation software for similarity analysis; (5) Multivariate statistical analysis: SPSS software was used to perform cluster analysis and principal component analysis on the common peak areas to evaluate the quality consistency between batches.

2. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (1), the specific preparation method of the test solution is as follows: weigh 0.5 g of the contents of Yangxin Dingji Capsules and place it in a conical flask, add 25 ml of 50% methanol solution, accurately add 25 mL of 50% methanol solution, weigh, ultrasonically treat, cool, weigh again, make up the lost amount with 50% methanol solution, shake well, filter, and filter the filtrate with a 0.45 μm microporous filter membrane to obtain.

3. The quality control method of Yangxin Dingji Capsule according to claim 2, characterized in that: The ultrasonic treatment was carried out at a power of 100 W and a frequency of 40 kHz for 30 min.

4. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (2), the liquid phase conditions include a flow rate of 1.0 mL min -1 ; Detection wavelength 265nm; injection volume 10μL; column temperature 30℃.

5. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (2), the gradient elution procedure is as follows:

6. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (3), the precision test uses chromatographic peak No. 7 as the reference peak, and calculates the RSD values ​​of the relative retention time and peak area of ​​each common peak, requiring RSD < 2%; The stability test was completed within 24 hours at room temperature, and the chromatographic peak No. 7 was used as the reference peak to calculate the relative retention time and peak area RSD values ​​of the common peaks, with the requirement that RSD < 2%; The repeatability test was performed by preparing 6 parallel test solutions, taking the chromatographic peak No. 7 as the reference peak, and calculating the RSD values ​​of the relative retention time and peak area of ​​each common peak, with the RSD requirement being <2%.

7. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (5), the cluster analysis adopts the inter-group linkage method and uses the squared Euclidean distance as the classification basis to divide the sample quality.

8. The quality control method of Yangxin Dingji Capsule according to claim 1, characterized in that: In step (5), the principal component analysis selects the principal components based on the condition that the eigenvalue is greater than 1.

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