Inspection method of traditional Chinese medicine spray
Through the combination of liquid chromatography and intelligent algorithms, the problem of insufficient accuracy and cumbersome operation of the detection of aconitum diester-type alkaloid content in traditional Chinese medicine sprays is solved, and efficient and accurate automated detection is achieved.
Patent Information
- Application Number
- CN202510735710.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-04
- Publication Date
- 2025-08-05
AI Technical Summary
The lack of specific aconitum diester-type alkaloid content detection methods in the prior art, resulting in insufficient detection accuracy and cumbersome operation.
Liquid chromatography was used, octadecylsilane bonded silica gel was used as filler, acetonitrile was mobile phase A, and 0.2% glacial acetic acid solution was mobile phase B. Gradient elution was used to establish a standard curve with support vector machine and partial least squares method. The chromatogram was analyzed using a convolutional neural network, and the purification process was controlled by the robotic arm to realize automated detection.
High accuracy detection of aconitum diester-type alkaloid content is achieved, the operation process is simplified, artificial errors are reduced, and detection efficiency and result reliability are improved.
Smart Images
Figure CN120427795A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of traditional Chinese medicine preparation detection, in particular to a method for detecting traditional Chinese medicine sprays. Background Art
[0002] Shuanghu Zhongtongning Spray is a traditional Chinese medicine spray that promotes qi circulation, reduces swelling and relieves pain, relaxes tendons and activates collaterals, and dispels wind and dampness. It is primarily composed of eight medicinal ingredients: Rhizoma Corydalis, Rhododendron azalea root, Radix Aconiti Kusnezoffii, Radix Aconiti Kusnezoffii, Rhizoma Pinelliae, Camphor, and Menthol. Radix Aconiti Kusnezoffii and Radix Aconiti Kusnezoffii primarily contain ester alkaloids, which have significant pharmacological effects but also carry a certain degree of toxicity. Therefore, accurate measurement of their content is crucial for controlling drug quality and ensuring drug safety.
[0003] However, the prior art does not have a specific method for detecting the content of aconitum diester-type alkaloids, resulting in problems of insufficient accuracy and complicated operation in the detection of aconitum diester-type alkaloids. Summary of the Invention
[0004] The present invention aims to provide a method for testing a Chinese medicine spray, aiming to solve the technical problems that the prior art lacks a specific method for detecting the content of aconitum diester-type alkaloids, resulting in insufficient accuracy and complicated operation in the detection of the content of aconitum diester-type alkaloids.
[0005] To achieve the above object, the present invention adopts a method for testing a Chinese medicine spray, comprising the following steps:
[0006] First, set the chromatographic conditions: the column is based on octadecylsilane bonded silica as the filler, acetonitrile or environmentally friendly solvent is used as mobile phase A, 0.2% glacial acetic acid solution is used as mobile phase B for gradient elution, and the detection wavelength is 235nm-280nm;
[0007] Prepare the control extract solution: take 20 mg of aconitine diester-type alkaloid control extract labeled with the content of mesaconitine, mesaconitine, and aconitine, accurately weigh it, place it in a 10 ml volumetric flask, dissolve it in the solution and dilute it to the mark, shake well to obtain the control extract solution;
[0008] A standard curve was established based on the control extract solution;
[0009] Perform sample testing based on the established standard curve: Take the Chinese medicine spray and filter it, then accurately draw 10μl, inject it into the liquid chromatograph, measure it based on the set chromatographic conditions, and complete the test by calculating according to the prepared standard curve.
[0010] The pH value of the glacial acetic acid solution used as mobile phase B was adjusted to 6.20 using triethylamine.
[0011] The aconitum diester type alkaloid control extract was placed in a volumetric flask and then dissolved and diluted with 0.05% hydrochloric acid ethanol solution.
[0012] The method for establishing a standard curve based on the control extract solution is as follows:
[0013] Accurately measure 1 ml of the control extract solution and place it in 2 ml, 5 ml, 10 ml, 25 ml, and 50 ml volumetric flasks, respectively. Add 0.01% hydrochloric acid methanol solution to dilute to the mark and shake well.
[0014] Accurately measure 10 μl of the control extract solution and the above-mentioned series of concentration control extract solutions, respectively, and inject them into the liquid chromatograph. Combined with the set chromatographic conditions, the determination is carried out, and the concentration of the control extract equivalent to metaaconitine, mesaconitine and aconitine is used as the horizontal axis, and the peak area value of the corresponding chromatographic peak is used as the vertical axis to establish a standard curve.
[0015] Among them, before starting the inspection method, the performance of the liquid chromatograph (HPLC) should be verified, including baseline noise, detection limit, quantification limit, and linear range parameters. At the same time, a convolutional neural network should be introduced to perform intelligent analysis of complex chromatograms.
[0016] During the test, mobile phase A and mobile phase B should be degassed before use.
[0017] Among them, when establishing the standard curve, the support vector machine (SVM) classification algorithm was introduced to preprocess the chromatogram to automatically identify and distinguish the target peaks from the impurity peaks; based on the partial least squares (PLS) regression analysis, a standard curve model of peak area and target component content was established.
[0018] Among them, based on the time series prediction algorithm (such as the ARIMA model), the historical maintenance data of the instrument is input, the instrument maintenance cycle is predicted, and maintenance reminders are generated.
[0019] Among them, the gradient elution method is as follows: introduce dynamic gradient elution technology, adjust the mobile phase ratio in real time according to the elution situation of the target component during the detection process, and improve the separation effect and detection speed.
[0020] The environmentally friendly solvent used as mobile phase A is an acetone-water mixture system.
[0021] Among them, take the traditional Chinese medicine spray and filter it, then accurately draw 10μl and inject it into the liquid chromatograph, the specific method is as follows:
[0022] A sample of Chinese medicine spray was taken, filtered through a 0.22 μm filter membrane, and then purified by a C18 solid phase extraction column. The target components were eluted with a methanol-water (70:30) solution.
[0023] The entire process is controlled by a robotic arm to reduce human errors.
[0024] At the same time, the data of pre-processing of traditional Chinese medicine spray samples are directly uploaded to the cloud and seamlessly connected to the HPLC system to realize the integrated process of "sample in - result out".
[0025] The invention discloses a method for testing a traditional Chinese medicine spray. When used, the method comprises the following steps: first, setting chromatographic conditions: a chromatographic column based on octadecylsilane bonded silica gel as a filler, using acetonitrile as mobile phase A, using 0.2% glacial acetic acid solution as mobile phase B for gradient elution, and detecting at a wavelength of 235 nm; then, taking 20 mg of aconitine diester-type alkaloid control extract labeled with the contents of neoaconitine, mesaconitine, and aconitine, accurately weighing the extract, placing the extract in a 10 ml volumetric flask, dissolving the extract with a solution, diluting the extract to the mark, and shaking the extract to obtain a control extract solution; establishing a standard curve based on the control extract solution; and conducting a sample test based on the established standard curve: taking the traditional Chinese medicine spray and filtering the extract, then accurately drawing 10 μl of the extract and injecting it into a liquid chromatograph, performing a measurement based on the set chromatographic conditions, and completing the test by calculating according to the prepared standard curve. This method solves the technical problems in the prior art of lacking a specific method for detecting the content of aconitine diester-type alkaloids, resulting in insufficient accuracy and complicated operation in detecting the content of aconitine diester-type alkaloids. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] 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 or the description of the prior art. 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.
[0027] Figure 1 It is a flow chart of the testing method of the Chinese medicine spray of the present invention. DETAILED DESCRIPTION
[0028] The embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, but should not be understood as limiting the present invention.
[0029] See also Figure 1 , Figure 1 It is a flow chart of the testing method of the Chinese medicine spray of the present invention.
[0030] The present invention provides a method for testing a Chinese medicine spray.
[0031] The steps include:
[0032] S1. First, set the chromatographic conditions: the chromatographic column is based on octadecylsilane bonded silica as the filler, acetonitrile or environmentally friendly solvent is used as mobile phase A, 0.2% glacial acetic acid solution is used as mobile phase B for gradient elution, and the detection wavelengths are 235nm-254nm and 280nm;
[0033] In this specific embodiment, the glacial acetic acid solution used as mobile phase B is adjusted to a pH of 6.20 using triethylamine. The primary wavelength can be set to 235 nm, while auxiliary wavelengths (such as 254 nm and 280 nm) are monitored simultaneously. Chromatographic data at multiple wavelengths is comprehensively processed using data analysis software to enhance the reliability of the detection results.
[0034] The gradient elution regulations are shown in the following table:
[0035] Table 1
[0036]
[0037] During the test, mobile phase A and mobile phase B should be degassed before use.
[0038] S2. Prepare a control extract solution: Take 20 mg of aconitine diester-type alkaloid control extract labeled with the content of mesaconitine, mesaconitine, and aconitine, accurately weigh it, place it in a 10 ml volumetric flask, dissolve it in the solution and dilute it to the mark, shake well to obtain the control extract solution;
[0039] According to this specific embodiment, the aconitum diester-type alkaloid control extract was placed in a volumetric flask and then dissolved and diluted with 0.05% hydrochloric acid ethanol solution.
[0040] S3. Establishing a standard curve based on the control extract solution;
[0041] For this specific embodiment, the method for establishing a standard curve based on the control extract solution is as follows:
[0042] Accurately measure 1 ml of the control extract solution and place it in 2 ml, 5 ml, 10 ml, 25 ml, and 50 ml volumetric flasks, respectively. Add 0.01% hydrochloric acid methanol solution to dilute to the mark and shake well.
[0043] Accurately measure 10 μl of the control extract solution and the above-mentioned series of concentration control extract solutions, respectively, and inject them into the liquid chromatograph. Combined with the set chromatographic conditions, the determination is carried out, and the concentration of the control extract equivalent to metaaconitine, mesaconitine and aconitine is used as the horizontal axis, and the peak area value of the corresponding chromatographic peak is used as the vertical axis to establish a standard curve.
[0044] When establishing the standard curve, the support vector machine (SVM) classification algorithm was introduced to preprocess the chromatogram to automatically identify and distinguish the target peaks from the impurity peaks. Based on the partial least squares (PLS) regression analysis, a standard curve model of peak area and target component content was established. At the same time, a convolutional neural network was introduced to perform intelligent analysis of complex chromatograms.
[0045] S4. Perform sample testing based on the established standard curve: Take the Chinese medicine spray and filter it, then accurately draw 10 μl and inject it into the liquid chromatograph. Perform the measurement based on the set chromatographic conditions and complete the test by calculating according to the prepared standard curve.
[0046] The test data is uploaded to the chain in real time to ensure that the test process is transparent and traceable.
[0047] Data on-chain: A hash value is generated for the chromatogram, standard curve parameters, inspection personnel information, etc. of each batch of samples and stored in a private chain node.
[0048] Anti-tampering mechanism: Any data modification requires verification by multiple nodes to ensure the authenticity of the test report.
[0049] Client query: Pharmaceutical companies or regulatory agencies can query full-process data through authorization keys.
[0050] Before starting the test method, the performance of the liquid chromatography (HPLC) instrument should be verified, including baseline noise, detection limit, quantification limit, and linear range parameters.
[0051] In actual application, based on the time series prediction algorithm, the patterns and trends of previous instrument maintenance data are mined. Through deep learning and analysis of the mined historical maintenance records (covering maintenance time intervals, maintenance items, instrument operating status changes and other multi-dimensional information), the maintenance cycle of the instrument is accurately predicted, and timely and accurate maintenance reminders are automatically generated based on the prediction results, effectively avoiding instrument failures due to improper or untimely maintenance, ensuring the long-term stable and efficient operation of the instrument, reducing the risk of equipment downtime and maintenance costs, and improving overall operational efficiency and reliability.
[0052] Among them, the gradient elution method is as follows: introduce dynamic gradient elution technology, adjust the mobile phase ratio in real time according to the elution situation of the target component during the detection process, and improve the separation effect and detection speed.
[0053] The environmentally friendly solvent used as mobile phase A is an acetone-water mixture system.
[0054] Among them, take the traditional Chinese medicine spray and filter it, then accurately draw 10μl and inject it into the liquid chromatograph, the specific method is as follows:
[0055] A sample of Chinese medicine spray was taken, filtered through a 0.22 μm filter membrane, and then purified by a C18 solid phase extraction column. The target components were eluted with a methanol-water (70:30) solution.
[0056] The entire process is controlled by a robotic arm to reduce human errors.
[0057] At the same time, the data of pre-processing of traditional Chinese medicine spray samples are directly uploaded to the cloud and seamlessly connected to the HPLC system to realize the integrated process of "sample in - result out".
[0058] In addition, the remote monitoring and diagnosis system enables remote monitoring, fault diagnosis and maintenance reminders for liquid chromatographs and other testing equipment.
[0059] Through the Internet of Things technology, the detection equipment is connected to the cloud server, and the equipment operation status data is uploaded in real time. Big data analysis technology is used to predict equipment failures and generate maintenance reminders in advance to ensure long-term and stable operation of the equipment.
[0060] The method for testing a traditional Chinese medicine spray according to the present embodiment is as follows: first, chromatographic conditions are set: a chromatographic column is filled with octadecylsilane bonded silica gel, acetonitrile is used as mobile phase A, 0.2% glacial acetic acid solution is used as mobile phase B for gradient elution, and the detection wavelength is 235 nm; then, 20 mg of aconitine diester-type alkaloid control extract labeled with the content of neoaconitine, mesaconitine, and aconitine is accurately weighed and placed in a 10 ml volumetric flask, dissolved with a solution and diluted to the scale, and shaken to obtain a control extract solution; a standard curve is established based on the control extract solution; and a sample is tested based on the established standard curve: the traditional Chinese medicine spray is taken and filtered, and then 10 μl is accurately aspirated and injected into a liquid chromatograph for measurement based on the set chromatographic conditions. The test is completed by calculation according to the prepared standard curve. This method solves the technical problem that the prior art lacks a specific method for detecting the content of aconitine diester-type alkaloids, resulting in insufficient accuracy and cumbersome operation in the detection of aconitine diester-type alkaloids.
[0061] The above disclosure is only a preferred embodiment of the present invention, and certainly cannot be used to limit the scope of the rights of the present invention. Ordinary technicians in this field can understand that all or part of the processes of the above embodiment and equivalent changes made in accordance with the claims of the present invention are still within the scope of the invention.
Claims
1. A method for testing a Chinese medicine spray, characterized in that: The steps include: First, set the chromatographic conditions: the column is based on octadecylsilane bonded silica as the filler, acetonitrile or environmentally friendly solvent is used as mobile phase A, 0.2% glacial acetic acid solution is used as mobile phase B for gradient elution, and the detection wavelength is 235nm-280nm; Prepare the control extract solution: take 20 mg of aconitine diester-type alkaloid control extract labeled with the content of mesaconitine, mesaconitine, and aconitine, accurately weigh it, place it in a 10 ml volumetric flask, dissolve it in the solution and dilute it to the mark, shake well to obtain the control extract solution; A standard curve was established based on the control extract solution; Sample testing is performed based on the established standard curve: the Chinese medicine spray is taken and filtered, and then 10 μl is accurately drawn and injected into the liquid chromatograph. The test is performed based on the set chromatographic conditions and calculated according to the prepared standard curve. Place the control extract of aconitum diester-type alkaloids in a volumetric flask, and dissolve and dilute it with 0.05% hydrochloric acid ethanol solution; The standard curve was established based on the control extract solution as follows: Accurately measure 1 ml of the control extract solution and place it in 2 ml, 5 ml, 10 ml, 25 ml, and 50 ml volumetric flasks, respectively. Add 0.01% hydrochloric acid methanol solution to dilute to the mark and shake well. Accurately measure 10 μl of the control extract solution and the above-mentioned series of concentration control extract solutions, respectively, and inject them into the liquid chromatograph. Combined with the set chromatographic conditions, the determination is carried out, and the concentration of the control extract equivalent to metaaconitine, mesaconitine and aconitine is used as the horizontal axis, and the peak area value of the corresponding chromatographic peak is used as the vertical axis to establish a standard curve.
2. The method for testing a Chinese medicine spray according to claim 1, wherein: The pH value of the glacial acetic acid solution used as mobile phase B was adjusted to 6.20 using triethylamine.
3. The method for testing a Chinese medicine spray according to claim 2, wherein: Before starting the test method, the performance of the liquid chromatography (HPLC) instrument should be verified, including baseline noise, detection limit, quantification limit, and linear range parameters.
4. The method for testing a Chinese medicine spray according to claim 3, wherein: During the test, mobile phase A and mobile phase B should be degassed before use.
5. The method for testing a Chinese medicine spray according to claim 4, wherein: When establishing the standard curve, the support vector machine (SVM) classification algorithm was introduced to preprocess the chromatogram to automatically identify and distinguish the target peaks from the impurity peaks. Based on the partial least squares (PLS) regression analysis, a standard curve model of peak area and target component content was established. At the same time, a convolutional neural network was introduced to perform intelligent analysis of complex chromatograms.
6. The method for testing a Chinese medicine spray according to claim 5, wherein: Based on the time series prediction algorithm, the historical maintenance data of the instrument is input to predict the instrument maintenance cycle and generate maintenance reminders.
7. The method for testing a Chinese medicine spray according to claim 1, wherein: The method for gradient elution is as follows: dynamic gradient elution technology is introduced to adjust the mobile phase ratio in real time according to the elution of the target component during the detection process, thereby improving the separation effect and detection speed.
8. The method for testing a Chinese medicine spray according to claim 1, wherein: The environmentally friendly solvent used as mobile phase A is an acetone-water mixture system.
9. The method for testing a Chinese medicine spray according to claim 1, wherein: Take the Chinese medicine spray and filter it, then accurately draw 10μl and inject it into the liquid chromatograph. The specific method is as follows: A sample of Chinese medicine spray was taken, filtered through a 0.22 μm filter membrane, and then purified by a C18 solid phase extraction column. The target components were eluted with a methanol-water (70:30) solution. The entire process is controlled by a robotic arm.