Rheum officinale quality determination method

Abstract

The invention relates to the technical field of medicinal material quality judgment, and discloses a rhubarb quality judgment method which comprises the following steps: S1, sample pretreatment; s2, anthraquinone component detection; S3, heavy metal and harmful element detection; and S4, data processing and quality determination. According to the method, a plurality of advanced instrument analysis technologies and data processing methods are combined, and comprehensive, rapid and accurate detection of the anthraquinone component content, the heavy metal content and the harmful element content in rheum officinale is realized. By optimizing technical details such as sample pretreatment, chromatographic conditions, gradient elution procedures, an online internal standard correction technology and a data processing model, the method not only improves the sensitivity and accuracy of detection, but also has the advantages of simplicity and convenience in operation and good repeatability, and provides powerful technical support for quality control and safety control of the rhubarb medicinal material.

Description

Technical Field

[0001] The invention relates to the technical field of medicinal material quality determination, and more particularly to a method for determining the quality of rhubarb. Background Art

[0002] As a traditional Chinese medicinal material, the quality of rhubarb directly affects its efficacy and clinical application effect. However, there are many shortcomings in the traditional methods for determining the quality of rhubarb. For example, the sample pretreatment process is cumbersome and time-consuming, and the digestion efficiency is low, resulting in low element recovery rate; the detection method of anthraquinone components has low sensitivity and poor accuracy, making it difficult to accurately determine total anthraquinones and specific anthraquinone components; the detection method of heavy metals and harmful elements has poor selectivity and is easily interfered by matrix effects, resulting in inaccurate measurement results; the data processing method is simple, lacks scientificity and systematicity, and it is difficult to fully and accurately reflect the quality characteristics of rhubarb.

[0003] Specifically, the existing methods for determining the quality of rhubarb usually use wet digestion or dry ashing for sample pretreatment. These methods are not only time-consuming, but also incomplete digestion, which can easily lead to element loss. In terms of the detection of anthraquinone components, although the traditional high performance liquid chromatography (HPLC) can achieve the separation and determination of anthraquinone components, it has low sensitivity and is easily interfered by other components. For the detection of heavy metals and harmful elements, the commonly used atomic absorption spectrometry (AAS) or atomic fluorescence spectrometry (AFS) has poor selectivity and is easily affected by matrix effects, resulting in inaccurate measurement results. In addition, most of the existing data processing methods use simple statistical methods, which lack scientificity and systematicity, and it is difficult to accurately analyze and determine the measurement results.

[0004] Therefore, there is an urgent need for an efficient and accurate method for determining the quality of rhubarb to solve the problems existing in the prior art. Summary of the invention

[0005] In view of this, the present invention provides an efficient and accurate method for determining the quality of rhubarb, which combines a variety of advanced instrumental analysis techniques and data processing methods to achieve comprehensive, rapid and accurate detection of anthraquinone components, heavy metals and harmful elements in rhubarb. By optimizing technical details such as sample pretreatment, chromatographic conditions, gradient elution procedures, online internal standard correction technology and data processing models, the present invention not only improves the sensitivity and accuracy of detection, but also has the advantages of simple operation and good repeatability, providing strong technical support for the quality control and safety control of rhubarb medicinal materials.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for determining the quality of rhubarb comprises the following steps:

[0008] S1. Sample pretreatment: The rhubarb sample was quickly digested by microwave digestion-acid removal method, and the excess acid was removed by acid removal step after the digestion was completed;

[0009] S2. Anthraquinone component detection: High performance liquid chromatography-diode array detector, combined with gradient elution program and column temperature control conditions, was used to accurately determine the total anthraquinones and specific anthraquinone components in rhubarb;

[0010] S3. Detection of heavy metals and harmful elements: Inductively coupled plasma mass spectrometry, combined with online internal standard correction technology and high-resolution mass spectrometry mode, as well as matrix matching correction, was used to determine the heavy metals and harmful elements of lead, cadmium, arsenic, mercury, and copper in rhubarb;

[0011] S4. Data processing and quality judgment: The multivariate linear regression model was used to process the measurement results. At the same time, the principal component analysis was combined to perform cluster analysis and outlier detection on the measurement results. The quality of rhubarb was judged based on the measurement results and quality judgment standards.

[0012] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the microwave digestion-acid-removing method, the microwave power is between 500W-500W, the digestion time is between 5 minutes and 30 minutes, the digestion temperature is between 120°C and 200°C, the acid-removing temperature is between 150°C and 250°C, and the acid-removing time is between 5 minutes and 20 minutes.

[0013] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the HPLC-diode array detector, the diode array detector is used for multi-wavelength detection of anthraquinone components.

[0014] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the gradient elution procedure, the initial mobile phase consists of methanol and water, wherein the initial volume percentage of methanol is set to 35%.

[0015] The gradient elution procedure is divided into three stages:

[0016] Phase 1: From the starting point, within 5 minutes, the volume percentage of methanol increases linearly to 50%, while the volume percentage of water decreases to 50% accordingly;

[0017] Stage 2: Subsequently, over the next 10 minutes, the volume percentage of methanol continued to increase linearly to 75%, while the volume percentage of water decreased to 25%;

[0018] Stage 3: Finally, in the remaining 5 minutes, the volume percentage of methanol was kept constant at 75% and the volume percentage of water was kept at 25% until the gradient elution program was completed.

[0019] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the column temperature control condition, the column temperature of the chromatographic column is set between 25°C and 35°C.

[0020] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the inductively coupled plasma mass spectrometry, the influence of instrument drift is eliminated by online internal standard correction technology, and the selectivity of the determination is improved by using a high-resolution mass spectrometry mode.

[0021] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the multivariate linear regression model, the independent variable is the content of each component in the measurement result, and the dependent variable is the content or quality index of the component to be measured. The regression equation is obtained by fitting and used for data processing.

[0022] Preferably, in the above-mentioned method for determining the quality of rhubarb, in the principal component analysis, the characteristic differences and outliers of rhubarb samples from different batches or sources are identified by performing dimensionality reduction processing and cluster analysis on the measurement results.

[0023] It can be seen from the above technical solution that, compared with the prior art, the present invention discloses a method for determining the quality of rhubarb, which has the following beneficial effects:

[0024] High sample pretreatment efficiency: Microwave digestion-acid-dripping method is used for sample pretreatment. By optimizing the power, time, temperature of microwave digestion and the temperature and time of the acid-dripping step, the digestion efficiency and element recovery rate are significantly improved, the sample loss and contamination are reduced, and an accurate and reliable sample basis is provided for subsequent detection.

[0025] Anthraquinone component detection has high sensitivity and good accuracy: High performance liquid chromatography-diode array detector (HPLC-DAD) is used to detect anthraquinone components, combined with gradient elution program and column temperature control conditions, to achieve accurate determination of total anthraquinone and specific anthraquinone components. The multi-wavelength detection function of the diode array detector improves the sensitivity and accuracy of the detection and effectively avoids interference from other components.

[0026] Good selectivity and high accuracy in heavy metal and harmful element detection: Inductively coupled plasma mass spectrometry (ICP-MS) is used to detect heavy metals and harmful elements, combined with online internal standard correction technology and high-resolution mass spectrometry mode, as well as matrix matching correction, which significantly improves the selectivity and accuracy of the determination. Online internal standard correction technology eliminates the influence of instrument drift, high-resolution mass spectrometry mode improves the selectivity of the determination, and matrix matching correction further eliminates the influence of matrix effects, ensuring the accuracy of the determination results.

[0027] Scientific and systematic data processing: The multivariate linear regression model is used to process the test results, and the principal component analysis (PCA) is combined to perform cluster analysis and outlier detection on the test results. This method not only improves the scientificity and systematicity of data processing, but also can comprehensively and accurately reflect the quality characteristics of rhubarb, providing a scientific basis for quality judgment. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0029] Figure 1 The accompanying drawing is a flow chart of the present invention. DETAILED DESCRIPTION

[0030] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] The embodiment of the present invention discloses a method for determining the quality of rhubarb, comprising the following steps:

[0032] S1. Sample pretreatment: The rhubarb sample was quickly digested by microwave digestion-acid removal method, and the excess acid was removed by acid removal step after the digestion was completed;

[0033] S2. Anthraquinone component detection: High performance liquid chromatography-diode array detector, combined with gradient elution program and column temperature control conditions, was used to accurately determine the total anthraquinones and specific anthraquinone components in rhubarb;

[0034] S3. Detection of heavy metals and harmful elements: Inductively coupled plasma mass spectrometry, combined with online internal standard correction technology and high-resolution mass spectrometry mode, as well as matrix matching correction, was used to determine the heavy metals and harmful elements of lead, cadmium, arsenic, mercury, and copper in rhubarb;

[0035] S4. Data processing and quality judgment: The multivariate linear regression model was used to process the measurement results. At the same time, the principal component analysis was combined to perform cluster analysis and outlier detection on the measurement results. The quality of rhubarb was judged based on the measurement results and quality judgment standards.

[0036] In the microwave digestion-acid removal method, the parameters such as the power, time, temperature of the microwave digestion and the temperature and time of the acid removal step are determined through the following optimization steps:

[0037] Microwave digestion parameter optimization:

[0038] Power selection: Under the premise of ensuring complete digestion of the sample, select a lower microwave power for digestion to reduce energy consumption and sample loss. Specifically, the microwave power is preferably in the range of 500W to 1500W, and the specific value is determined according to the sample amount and the material of the digestion container.

[0039] Time control: The digestion time is determined according to factors such as the type of sample, water content, and size of the digestion container. While ensuring complete digestion, try to shorten the digestion time to improve work efficiency. The digestion time is preferably in the range of 5 minutes to 30 minutes.

[0040] Temperature setting: Digestion temperature is a key factor affecting digestion efficiency and element recovery rate. Under the premise of avoiding sample boiling and container rupture, try to increase the digestion temperature to improve the digestion speed and element recovery rate. The digestion temperature is preferably in the range of 120℃ to 200℃.

[0041] Optimization of parameters in the acid-chasing step:

[0042] Temperature selection: The acid removal temperature should be higher than the digestion temperature to promote the volatilization and removal of excess acid. At the same time, excessive temperature should be avoided to cause sample decomposition or container rupture. The acid removal temperature is preferably in the range of 150°C to 250°C.

[0043] Time control: The acid removal time should be determined according to the type and concentration of the acid in the digestion solution and the amount of sample. Under the premise of ensuring that the excess acid is completely removed, the acid removal time should be shortened as much as possible to reduce sample loss and contamination. The acid removal time is preferably within the range of 5 to 20 minutes.

[0044] In order to further optimize the above technical solution, in the high performance liquid chromatography-diode array detector, the diode array detector is used to perform multi-wavelength detection of the anthraquinone components.

[0045] In order to further optimize the above technical solution, in the gradient elution program, the initial mobile phase consists of methanol and water, wherein the initial volume percentage of methanol is set to 35%.

[0046] The gradient elution procedure is divided into three stages:

[0047] Phase 1: From the starting point, within 5 minutes, the volume percentage of methanol increases linearly to 50%, while the volume percentage of water decreases to 50% accordingly;

[0048] Stage 2: Subsequently, over the next 10 minutes, the volume percentage of methanol continued to increase linearly to 75%, while the volume percentage of water decreased to 25%;

[0049] Stage 3: Finally, in the remaining 5 minutes, the volume percentage of methanol was kept constant at 75% and the volume percentage of water was kept at 25% until the gradient elution program was completed.

[0050] In order to further optimize the above technical solution, the column temperature of the chromatographic column was set between 25°C and 35°C to ensure that the anthraquinone components in rhubarb had the best separation effect and stability during the analysis process.

[0051] In order to further optimize the above technical solution, in the inductively coupled plasma mass spectrometry, the influence of instrument drift is eliminated by online internal standard correction technology, and the selectivity of the determination is improved by using high-resolution mass spectrometry mode.

[0052] In order to further optimize the above technical solution, in the multivariate linear regression model, the independent variable is the content of each component in the measurement results, and the dependent variable is the content or quality index of the component to be measured. The regression equation is obtained by fitting and used for data processing.

[0053] In order to further optimize the above technical solution, in principal component analysis, the characteristic differences and outliers of rhubarb samples from different batches or sources were identified by performing dimensionality reduction and cluster analysis on the measurement results.

[0054] Technical principle:

[0055] Sample pretreatment (microwave digestion-acid removal method):

[0056] Principle: Microwave energy is used to quickly heat the sample and react with acid to quickly digest the sample. After digestion is completed, excess acid is removed by heating (driving away acid) to facilitate subsequent accurate analysis. Microwave digestion has the advantages of uniform heating, rapid heating and energy saving.

[0057] Parameter optimization: By setting microwave power, digestion time, digestion temperature, and acid removal temperature and time, ensure that the sample is fully digested without affecting subsequent analysis.

[0058] Anthraquinone component detection (HPLC-diode array detector):

[0059] Principle: HPLC is used to separate the anthraquinone components in rhubarb, and a diode array detector is used for multi-wavelength detection to improve the accuracy and sensitivity of the detection. The gradient elution procedure achieves the separation of components of different polarity by changing the composition of the mobile phase.

[0060] Parameter optimization: By setting the composition of the initial mobile phase and the gradient elution program, as well as the temperature of the chromatographic column, effective separation and accurate detection of anthraquinone components were ensured.

[0061] Heavy metal and harmful element detection (inductively coupled plasma mass spectrometry):

[0062] Principle: Inductively coupled plasma mass spectrometry is used to convert heavy metals and harmful elements in samples into ions and detect them through a mass spectrometer. Online internal standard correction technology is used to eliminate the influence of instrument drift, and high-resolution mass spectrometry mode improves the selectivity of the determination.

[0063] Parameter Optimization: Ensure the accuracy and reliability of the measurement by optimizing instrument parameters and using matrix matching correction.

[0064] Data processing and quality assessment (multiple linear regression model and principal component analysis):

[0065] Principle: The multivariate linear regression model is used to fit the relationship between the test results and the component content, and the regression equation is obtained for data processing. The principal component analysis identifies the characteristic differences and outliers of rhubarb samples from different batches or sources through dimensionality reduction and cluster analysis.

[0066] Application: The quality of rhubarb is determined based on the test results and quality judgment criteria, combined with the results of multiple linear regression model and principal component analysis.

[0067] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the method part.

[0068] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for determining the quality of rhubarb, characterized in that: The following steps are involved: S1. Sample pretreatment: The rhubarb sample was quickly digested by microwave digestion-acid removal method, and the excess acid was removed by acid removal step after the digestion was completed; S2. Anthraquinone component detection: High performance liquid chromatography-diode array detector, combined with gradient elution program and column temperature control conditions, was used to accurately determine the total anthraquinones and specific anthraquinone components in rhubarb; S3. Detection of heavy metals and harmful elements: Inductively coupled plasma mass spectrometry, combined with online internal standard correction technology and high-resolution mass spectrometry mode, as well as matrix matching correction, was used to determine the heavy metals and harmful elements of lead, cadmium, arsenic, mercury, and copper in rhubarb; S4. Data processing and quality judgment: The multivariate linear regression model was used to process the measurement results. At the same time, the principal component analysis was combined to perform cluster analysis and outlier detection on the measurement results. The quality of rhubarb was judged based on the measurement results and quality judgment standards.

2. A method for determining the quality of rhubarb according to claim 1, characterized in that: In the microwave digestion-acid removal method, the microwave power is between 500W-500W, the digestion time is between 5 minutes and 30 minutes, the digestion temperature is between 120°C and 200°C, the acid removal temperature is between 150°C and 250°C, and the acid removal time is between 5 minutes and 20 minutes.

3. A method for determining the quality of rhubarb according to claim 1, characterized in that: In the high performance liquid chromatography-diode array detector, the diode array detector is used to perform multi-wavelength detection on the anthraquinone components.

4. The method for determining the quality of rhubarb according to claim 1, characterized in that: In the gradient elution procedure, the initial mobile phase consists of methanol and water, wherein the initial volume percentage of methanol is set to 35%; The gradient elution procedure is divided into three stages: Phase 1: From the starting point, within 5 minutes, the volume percentage of methanol increases linearly to 50%, while the volume percentage of water decreases to 50% accordingly; Stage 2: Subsequently, over the next 10 minutes, the volume percentage of methanol continued to increase linearly to 75%, while the volume percentage of water decreased to 25%; Stage 3: Finally, in the remaining 5 minutes, the volume percentage of methanol was kept constant at 75% and the volume percentage of water was kept at 25% until the gradient elution program was completed.

5. The method for determining the quality of rhubarb according to claim 1, characterized in that: In the column temperature control condition, the column temperature of the chromatographic column is set between 25°C and 35°C.

6. The method for determining the quality of rhubarb according to claim 1, characterized in that: In the inductively coupled plasma mass spectrometry, the influence of instrument drift is eliminated by online internal standard correction technology, and the selectivity of the determination is improved by using a high-resolution mass spectrometry mode.

7. The method for determining the quality of rhubarb according to claim 1, characterized in that: In the multivariate linear regression model, the independent variable is the content of each component in the measurement result, and the dependent variable is the content or quality index of the component to be measured. The regression equation is obtained by fitting and used for data processing.

8. The method for determining the quality of rhubarb according to claim 1, characterized in that: In the principal component analysis, the characteristic differences and outliers of rhubarb samples from different batches or sources are identified by performing dimensionality reduction processing and cluster analysis on the measurement results.

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