Chromatograph mobile phase adaptive proportioning method and system

Abstract

This application relates to an adaptive mobile phase ratio method and system for chromatographs, specifically in the field of chromatograph elution control. The method includes acquiring preset fixed elution parameters and baseline elution data for the components to be separated; conducting a pre-elution experiment based on the fixed elution parameters to determine the current elution data; analyzing the current elution data and baseline elution data to determine the overall and partial offset indices; analyzing the overall and partial offset indices to determine fixed compensation parameters; and compensating the fixed elution parameters based on the fixed compensation parameters to determine adaptive elution parameters. This application improves the elution efficiency of the chromatograph.

Description

Technical Field

[0001] This application relates to the technical field of elution control in chromatographs, and in particular to an adaptive mobile phase ratio method and system for chromatographs. Background Technology

[0002] Adaptive mobile phase ratio for chromatographs refers to the process of optimizing the elution effect of the chromatograph by changing the mobile phase ratio parameters, and then eluting biomass energy components based on the optimized mobile phase ratio.

[0003] In related technologies, when preparing the mobile phase for a chromatograph, a fixed ratio is usually adopted. By setting different mobile phase ratios, offline pre-experiments are conducted on the substances to be eluted. The mobile phase ratio parameter with the best elution effect is selected as the final elution ratio parameter. The elution ratio parameter is then fixed and used to elute the corresponding biomass energy substances to be eluted.

[0004] Regarding the aforementioned technologies, when setting a fixed mobile phase to elute biomass energy materials, due to differences in model or service life between different chromatographs, the elution effect of different chromatographs under fixed parameters may vary, and the fixed elution ratio parameters may not match the current chromatograph parameters, resulting in low elution efficiency of the chromatograph and room for improvement. Summary of the Invention

[0005] To improve the elution efficiency of a chromatograph, this application provides a method and system for adaptive mobile phase ratio matching for a chromatograph.

[0006] In a first aspect, this application provides an adaptive mobile phase ratio method for chromatographs, employing the following technical solution: An adaptive mobile phase mixing method for chromatographs includes: Obtain the preset fixed elution parameters and baseline elution data of the components to be separated; A preliminary elution experiment was conducted on the components to be separated using fixed elution parameters to determine the current elution data. The current elution data and the baseline elution data are analyzed to determine the overall offset index and the partial offset index; The overall offset index and partial offset index are analyzed to determine the fixed compensation parameters; The fixed elution parameters are compensated based on the fixed compensation parameters to determine the adaptive elution parameters.

[0007] Optionally, the steps of analyzing the current elution data and baseline elution data to determine the overall and partial offset indices include: Calculate the average value of the current elution data to determine the average measured data; Determine whether the average measured data meets the requirements of the baseline elution data; If so, the preset unbiased global index and the preset unbiased partial index are respectively determined as the global offset index and the partial offset index. If not, the average measured data and the baseline elution data are analyzed to determine the overall offset index and the partial offset index.

[0008] Optionally, the steps of analyzing the average measured data and baseline elution data to determine the overall and partial offset indices include: Data extraction was performed on the average measured data to determine the mean dead time, the mean chromatographic peak time, and the reference dead time; Calculate the relative proportion of the difference between the mean time value of the chromatographic peak and the mean dead time value to determine the measured capacity factor; Data extraction was performed on the baseline elution data to determine the baseline chromatographic time; Calculate the relative proportion of the difference between the baseline chromatographic time and the mean of the baseline dead time to determine the baseline capacity factor; The measured capacity factor and the baseline capacity factor are analyzed to determine the overall offset index and the partial offset index.

[0009] Optionally, the steps of analyzing the measured capacity factor and the baseline capacity factor to determine the overall offset index and the partial offset index include: Calculate the quotient between the measured capacity factor and the baseline capacity factor to determine the measured shrinkage index; Numerical analysis of the measured contraction index was performed to determine the maximum and minimum contraction index. Calculate the quotient of the minimum contraction index and the maximum contraction index to determine the overall offset index; Calculate the absolute difference between the measured capacity factor and the reference capacity factor to determine the peak relative deviation; Numerical analysis of the relative peak deviation was performed to determine the maximum and minimum peak deviations. Calculate the difference between the maximum peak deviation and the minimum peak deviation to determine the partial offset index.

[0010] Optionally, the steps of analyzing the overall offset index and partial offset index to determine the fixed compensation parameters include: Obtain the percentage of actual time and the percentage of baseline time; Determine whether the overall offset index is greater than the preset overall offset threshold; If it is greater than that, the proportion of measured time and the proportion of reference time will be analyzed to determine the fixed compensation parameters; If it is not greater than, then determine whether the partial offset index is greater than the preset partial offset threshold. If it is greater than the baseline time ratio, the absolute deviation between the measured time ratio and the baseline time ratio is calculated to determine the measured ratio deviation. The measured percentage deviation, measured time percentage, and reference time percentage are analyzed to determine the fixed compensation parameters; If it is not greater than, then the preset zero compensation parameter will be determined as the fixed compensation parameter.

[0011] Optionally, the steps to analyze the proportion of measured time and the proportion of reference time to determine the fixed compensation parameters include: Calculate the relative deviation between the measured time percentage and the reference time percentage to determine the time percentage deviation; Calculate the mean of the time percentage deviation to determine the mean of the percentage deviation; Calculate the product of the average percentage deviation and the preset compensation adjustment factor to determine the initial overall compensation amount; Elution pre-experiments were conducted on the substances to be eluted based on the initial overall compensation amount and the preset fine-tuning upper limit to determine the fixed compensation parameters.

[0012] Optionally, the steps to analyze the measured percentage deviation, measured time percentage, and reference time percentage to determine the fixed compensation parameters include: Numerical analysis was performed on the measured percentage deviation to determine the maximum percentage deviation. Data on the baseline time percentage is extracted based on the maximum percentage deviation to determine the target baseline percentage; Calculate the quotient of the maximum percentage deviation and the target baseline percentage to determine the target relative deviation; Calculate the product of the target relative deviation and the preset compensation adjustment factor to determine the initial partial compensation amount; A preliminary elution experiment was conducted on the substance to be eluted based on the initial partial compensation amount and the preset upper limit for fine-tuning, in order to determine the fixed compensation parameters.

[0013] Secondly, this application provides an adaptive mobile phase mixing system for a chromatograph, which adopts the following technical solution: An adaptive mobile phase mixing system for a chromatograph, comprising: The acquisition module is used to acquire fixed elution parameters and baseline elution data; A memory for storing a program for an adaptive chromatograph mobile phase ratio method as described in any of the preceding claims; The processor and the program in the memory can be loaded and executed by the processor to implement an adaptive ratio method for the mobile phase of a chromatograph as described in any of the above.

[0014] In summary, this application includes at least one of the following beneficial technical effects: 1. By acquiring fixed elution parameters and baseline elution data for the components to be separated, a preliminary elution experiment is conducted based on the fixed elution parameters to determine the current elution data. The current elution data and baseline elution data are analyzed to determine the overall and partial offset indices. The overall and partial offset indices are then analyzed to determine the fixed compensation parameters. The fixed elution parameters are then compensated based on the fixed compensation parameters to determine the adaptive elution parameters. This allows for the analysis of chromatograms that do not meet the baseline data requirements. When an overall offset occurs in the chromatogram, it indicates that all eluted components in the system have undergone proportional and unidirectional global scaling. Therefore, the organic phase ratio in the mobile phase should be uniformly compensated first to bring the elution results to the standard state. When a partial offset occurs in the chromatogram, it indicates that the system has undergone systemic changes such as gradient delay volume changes, mobile phase pH deviations, or column temperature drift. Therefore, the mobile phase ratio is targeted and compensated within a small range to reduce interference to other components while compensating for partial anomalies, thereby optimizing the elution effect of the chromatograph and improving the accuracy of biomass energy component detection. 2. By extracting data from the average measured data, the mean dead time, mean chromatographic peak time, and reference dead time are determined. The relative proportion of the difference between the mean chromatographic peak time and the mean dead time is calculated to determine the measured capacity factor. Data is extracted from the reference elution data to determine the reference chromatographic time. The relative proportion of the difference between the reference chromatographic time and the mean reference dead time is calculated to determine the reference capacity factor. The measured capacity factor and the reference capacity factor are analyzed to determine the overall shift index and the partial shift index. Thus, based on the consistency of the scaling ratio of the retention time of all chromatographic peaks relative to the reference peak during operation, the overall shift of the chromatogram relative to the corresponding reference state is evaluated, improving the accuracy of the mobile phase adaptive ratio of the chromatograph. 3. By numerically analyzing the measured proportion deviation, the maximum proportion deviation is determined. Based on the maximum proportion deviation, the baseline time proportion is extracted to determine the target baseline proportion. The quotient of the maximum proportion deviation and the target baseline proportion is calculated to determine the target relative deviation. The product of the target relative deviation and the preset compensation adjustment factor is calculated to determine the initial partial compensation amount. Based on the initial partial compensation amount and the preset fine-tuning upper limit, a pre-elution experiment is conducted on the eluted substance to determine the fixed compensation parameters. Thus, different compensation adjustment factors are selected according to different chromatographic separation modes, and the mobile phase ratio is adjusted for the target baseline. This ensures the visualization and traceability of the adjustment process, avoids black-box adjustment, and improves the efficiency and standardization of mobile phase adaptive ratio adjustment in the chromatograph. Attached Figure Description

[0015] Figure 1 This is a flowchart of an adaptive mobile phase ratio method for chromatographs according to an embodiment of this application.

[0016] Figure 2 This is a flowchart illustrating the analysis of current elution data and baseline elution data in this application embodiment to determine the overall offset index and partial offset index.

[0017] Figure 3 This is a flowchart illustrating the analysis of average measured data and baseline elution data in this application embodiment to determine the overall offset index and partial offset index.

[0018] Figure 4 This is a flowchart illustrating the analysis of measured capacity factors and baseline capacity factors in this application embodiment to determine the overall offset index and partial offset index.

[0019] Figure 5 This is a flowchart illustrating the analysis of the overall offset index and partial offset index in this embodiment of the application to determine the fixed compensation parameters.

[0020] Figure 6 This is a flowchart in this application embodiment that analyzes the proportion of measured time and the proportion of reference time to determine the fixed compensation parameters.

[0021] Figure 7 This is a flowchart in this application embodiment that analyzes the measured percentage deviation, measured time percentage, and reference time percentage to determine the fixed compensation parameters. Detailed Implementation

[0022] To make the purpose, technical solution, and advantages of this application clearer, the following description is provided in conjunction with the appendix. Figures 1 to 7 The present application will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the application.

[0023] This application discloses a method and system for adaptive mobile phase ratio in chromatography. Specifically, it discloses a processing terminal that acquires fixed elution parameters and reference elution parameters. Based on the fixed elution parameters, a pre-elution experiment is performed on the components to be separated to determine the current elution data. The current elution data and reference elution data are analyzed to determine the overall and partial offset indices. Further analysis of the overall and partial offset indices determines fixed compensation parameters. The fixed elution parameters are then compensated based on these fixed compensation parameters to determine adaptive elution parameters, thereby correcting chromatograms that do not meet the reference data requirements. Analysis revealed that when a chromatogram shows a global shift, it indicates that all eluted components in the system have undergone proportional and unidirectional global scaling. Therefore, in this case, the organic phase ratio in the mobile phase should be compensated uniformly to bring the elution results back to the standard state. When a chromatogram shows a partial shift, it indicates that the system has undergone systemic changes such as gradient delay volume changes, mobile phase pH deviations, or column temperature drift. Therefore, in this case, the mobile phase ratio should be compensated for on a small scale to reduce interference to other components while compensating for the partial anomalies, thereby optimizing the elution effect of the chromatograph and improving the accuracy of biomass energy component detection.

[0024] Reference Figure 1 This application discloses an adaptive mobile phase ratio method for chromatographs, comprising the following steps: Step S100: Obtain the preset fixed elution parameters and baseline elution data of the components to be separated.

[0025] The component to be separated refers to a mixed biomass energy sample component that is to be eluted by the chromatograph and has one or more effective eluting components. It is a known calibration component, such as n-alkanes or uracil, which does not affect the chemical properties of the eluting substance. The effective eluting components in the preliminary experiment include an introduced internal standard. The component to be separated with the introduced calibration component is used to compensate for the deviation of the chromatograph in the preliminary experiment. After compensation, no calibration component is introduced when the component to be separated is eluted for formal separation.

[0026] Fixed elution parameters refer to the fixed chromatographic elution control parameters that have been developed and validated for the components to be separated. These include fixed operating parameters such as chromatographic mobile phase ratio, mobile phase flow rate, and column temperature. These parameters are determined by the processing terminal by searching the standard detection database stored in the system based on the properties of the components to be separated.

[0027] Reference elution data refers to the SST standard range of the main eluted components in the chromatograph, used to verify the effectiveness of the system separation. It includes the peak retention time range, lower limit of critical peak resolution, main peak tailing factor range, lower limit of theoretical plate number, repeatability range, spiked recovery rate range, and peak symmetry factor range of the effective eluted components in the substance when eluting. It is determined by the processing terminal by looking up the main eluted components in the standard detection database stored in the system.

[0028] Step S101: Perform a preliminary elution experiment on the component to be separated according to the fixed elution parameters to determine the current elution data.

[0029] The current elution data refers to the elution parameter data obtained after performing elution pre-experiments on the components to be separated, including the elution dead time of the components to be eluted, the peak retention time of the effective eluted components, the resolution of the key peak, the tailing factor of the main peak, the theoretical plate number, repeatability, the peak symmetry factor, and the spiked recovery rate. The processing terminal first retrieves multiple sets of chromatogram data obtained after performing multiple elution pre-experiments on the components to be separated according to fixed elution parameters, then analyzes the chromatograms, and calculates the peak retention time of the main eluted component, the resolution of the key peak, the tailing factor of the main peak, the theoretical plate number, repeatability, the peak symmetry factor, the spiked recovery rate, and the elution dead time of each pre-experiment. When performing elution pre-experiments, the operator ensures that there are no hardware malfunctions in the chromatograph, that it has undergone a power-on self-test, that all components of the chromatograph are within their normal operating cycle, that the mobile phase of the chromatograph is within its validity period and is properly prepared according to the fixed elution parameters, and that accidental operational errors are excluded.

[0030] Step S102: Analyze the current elution data and the baseline elution data to determine the overall offset index and the partial offset index.

[0031] The overall offset index is a quantitative indicator of the overall offset of the chromatogram determined in the preliminary experimental run relative to the baseline elution data. The partial offset index is a quantitative indicator of the partial offset of the chromatogram determined in the preliminary experimental run relative to the baseline elution data. This allows for the analysis of chromatograms that do not meet the baseline data requirements. When an overall offset occurs, it indicates that all eluted components in the system have undergone proportional and unidirectional global scaling. Therefore, priority should be given to uniformly compensating the organic phase ratio in the mobile phase to bring the elution results to the standard state. When a partial offset occurs, it indicates that the system has experienced systemic changes such as gradient delay volume changes, mobile phase pH deviations, or column temperature drift. Therefore, small-scale targeted compensation of the mobile phase ratio is performed to compensate for partial anomalies while reducing interference to other components, thus optimizing the elution effect of the chromatograph. Both are determined by the processing terminal through analysis of the current elution data and the baseline elution data. Specific analysis steps are described in [reference needed]. Figure 2 The steps in the process.

[0032] Step S103: Analyze the overall offset index and partial offset index to determine the fixed compensation parameters.

[0033] The fixed compensation parameter refers to the fixed mobile phase ratio compensation parameter corresponding to the overall or partial shift of the chromatographic elution results relative to the reference elution data. It is determined by the processing terminal through analysis of the overall and partial shift indices. Specific analysis steps are described in [reference needed]. Figure 5 The steps in the process.

[0034] Step S104: Compensate the fixed elution parameters according to the fixed compensation parameters to determine the adaptive elution parameters.

[0035] The adaptive elution parameter refers to the fixed mobile phase ratio parameter after being compensated by the fixed compensation parameter. After determining the adaptive elution parameter, the biomass sample is eluted based on the adaptive elution parameter, thereby improving the accuracy of biomass energy-related detection. The fixed elution parameter is determined by the processing terminal after determining the fixed compensation parameter and then correcting the fixed elution parameter based on the fixed compensation parameter.

[0036] Reference Figure 2 The steps for analyzing current elution data and baseline elution data to determine the overall and partial offset indices include: Step S200: Calculate the average value of the current elution data to determine the average measured data.

[0037] The average measured data refers to the average value of the current elution data corresponding to multiple sets of elution pre-experiments. It is determined by the processing terminal by calculating the average value of the current elution data, thereby reducing the impact of external disturbances on the experimental results and improving the accuracy of the mobile phase adaptive ratio of the chromatograph.

[0038] Step S201: Determine whether the average measured data meets the requirements of the baseline elution data.

[0039] The requirement for baseline elution data is that all parameters in the average measured data are within the range of the baseline elution data.

[0040] By processing the terminal to determine whether the average measured data are all within the range of the reference elution data, it can be determined whether the elution results of the chromatograph are within the range of the standard data, whether the mobile phase of the chromatograph needs to be fixed and compensated, and thus optimize the operation of the chromatograph.

[0041] Step S2011: If so, then the preset unbiased global index and the preset unbiased partial index are respectively determined as the global offset index and the partial offset index.

[0042] If the processing terminal determines that all data items in the average measured data fall within the range of the reference elution data, it indicates that the elution results of the chromatograph are within the range of the standard data and there is no need to perform fixed compensation on the ratio of the mobile phase of the chromatograph. Therefore, the unbiased overall index and the unbiased partial index are determined as the overall offset index and the partial offset index, respectively.

[0043] The unbiased global index is a global offset index used to calibrate the system to indicate that no global offset has occurred. It is set to 1 and is calibrated by the operator in the system.

[0044] The unbiased partial index is a partial offset index used to calibrate the system without partial offset. It is set to 0 and is calibrated by the operator in the system.

[0045] The overall offset index is consistent with the overall offset index in step S102. It is determined by the processing terminal based on the unbiased overall index when the elution results of the chromatograph are within the standard data range and there is no need to perform fixed compensation on the ratio of the mobile phase of the chromatograph.

[0046] The partial offset index is consistent with the overall offset index in step S102. It is determined by the processing terminal based on the unbiased partial index when the elution results of the chromatograph are within the standard data range and there is no need to perform fixed compensation on the ratio of the mobile phase of the chromatograph.

[0047] Step S2012: If not, analyze the average measured data and the baseline elution data to determine the overall offset index and the partial offset index.

[0048] If the processing terminal determines that there are data items in the average measured data that do not fall within the range of the reference elution data, it indicates that the elution results of the chromatograph exceed the standard data range, and there is a deviation in the elution results due to aging or changes in the chromatograph model. Therefore, the average measured data and the reference elution data are analyzed to determine the overall offset index and the partial offset index, which provides data support for the subsequent determination of fixed compensation parameters, thereby optimizing the elution effect of the chromatograph.

[0049] The overall offset index is consistent with the overall offset index in step S102. The partial offset index is consistent with the partial offset index in step S102. Both are determined by the processing terminal after determining that the elution results of the chromatograph exceed the standard data range, indicating aging or deviation due to changes in the chromatograph model. The analysis involves comparing the average measured data and the baseline elution data. Specific analysis steps are described below. Figure 3 The steps in the process.

[0050] Reference Figure 3The steps for analyzing average measured data and baseline elution data to determine the overall and partial offset indices include: Step S300: Extract data from the average measured data to determine the mean dead time, the mean chromatographic peak time, and the baseline dead time.

[0051] The mean dead time refers to the average elution dead time in the average measured data, which is determined by the processing terminal by extracting the mean elution dead time from the average measured data.

[0052] The mean peak time in chromatography refers to the peak retention time of each effective eluting component in the average measured data. It is determined by the processing terminal through data extraction of the mean peak retention time of each effective eluting component in the average measured data.

[0053] The baseline dead time refers to the elution dead time corresponding to the baseline elution data, which is determined by the processing terminal by extracting the average elution dead time from the average measured data.

[0054] Step S301: Calculate the relative proportion of the difference between the mean time of the chromatographic peak and the mean dead time to determine the measured capacity factor.

[0055] Among them, the measured capacity factor refers to the capacity factor corresponding to each effective elution component in the pre-experiment process. It is determined by the processing terminal by first calculating the difference between the mean dead time and the mean chromatographic peak time, and then calculating the quotient of the time difference and the mean dead time.

[0056] Step S302: Extract data from the reference elution data to determine the reference chromatographic time.

[0057] The reference chromatographic time refers to the reference peak retention time corresponding to each effective eluted component. It is determined by the processing terminal by first extracting the peak retention time range of each effective eluted component in the reference elution data, and then taking the median value of the peak retention time range.

[0058] Step S303: Calculate the relative proportion of the difference between the baseline chromatographic time and the mean baseline dead time to determine the baseline capacity factor.

[0059] The baseline capacity factor refers to the capacity factor corresponding to each effective eluting component under the baseline elution data. It is determined by the processing terminal by first calculating the difference between the mean baseline dead time and the baseline chromatographic time, and then calculating the quotient of the time difference and the baseline dead time.

[0060] Step S304: Analyze the measured capacity factor and the baseline capacity factor to determine the overall offset index and the partial offset index.

[0061] The overall offset index is consistent with the overall offset index in step S2012. The partial offset index is consistent with the partial offset index in step S2012. Both are determined by the processing terminal through analysis of the measured capacity factor and the reference capacity factor. Specific analysis steps are detailed below. Figure 4 The steps in the process.

[0062] Reference Figure 4 The steps for analyzing the measured capacity factor and the baseline capacity factor to determine the overall offset index and the partial offset index include: Step S400: Calculate the quotient of the measured capacity factor and the baseline capacity factor to determine the measured shrinkage index.

[0063] The measured shrinkage index refers to the shrinkage ratio of the measured capacity factor of each effective elution component relative to the baseline capacity factor, which is determined by the processing terminal by calculating the quotient of the measured capacity factor and the baseline capacity factor.

[0064] Step S401: Perform numerical analysis on the measured contraction index to determine the maximum and minimum contraction index.

[0065] The maximum contraction index refers to the maximum value among the measured contraction indices. It is determined by traversing the measured contraction indices to find the maximum value, which is the maximum contraction index.

[0066] The minimum contraction index refers to the minimum value among the measured contraction indices. The processing terminal determines the minimum value of the measured contraction index by iterating through the measured contraction indices.

[0067] Step S402: Calculate the quotient of the minimum contraction index and the maximum contraction index to determine the overall offset index.

[0068] The overall offset index is consistent with the overall offset index in step S304. It is determined by the processing terminal by calculating the quotient of the minimum shrinkage index and the maximum shrinkage index. In this way, the overall offset of the chromatogram relative to the corresponding reference state is evaluated based on the consistency of the scaling ratio of the retention time of all chromatographic peaks relative to the reference peak during the operation, thereby improving the accuracy of the mobile phase adaptive ratio of the chromatograph.

[0069] Step S403: Calculate the absolute difference deviation between the measured capacity factor and the reference capacity factor to determine the peak relative deviation.

[0070] Among them, peak relative deviation refers to the deviation of the measured capacity factor of the effective elution component from the reference capacity factor. It is determined by the processing terminal by first calculating the average value of the difference between the measured capacity factor and the reference capacity factor, and then calculating the quotient of the average value of the difference and the measured capacity factor.

[0071] Step S404: Perform numerical analysis on the relative peak deviation to determine the maximum and minimum peak deviation.

[0072] The maximum peak deviation refers to the maximum value among the relative peak deviations. The processing terminal determines the maximum value among the relative peak deviations by traversing the relative peak deviations.

[0073] Minimum peak deviation refers to the minimum value among peak relative deviations. The processing terminal determines the minimum value among the peak relative deviations by traversing the peak relative deviations.

[0074] Step S405: Calculate the difference between the maximum peak deviation and the minimum peak deviation to determine the partial offset index.

[0075] The partial offset index is consistent with the partial offset index in step S304. It is determined by the processing terminal by calculating the difference between the maximum peak deviation and the minimum peak deviation. Based on the maximum difference in the relative offset amplitude of the capacity factor of each effective elution component relative to the corresponding reference capacity factor, the deviation between the offset degree of different effective elution components relative to the reference state is determined, and the partial offset degree between each effective elution component is quantitatively determined, thereby improving the accuracy of the mobile phase ratio of the chromatograph.

[0076] Reference Figure 5 The steps for analyzing the overall offset index and partial offset index to determine the fixed compensation parameters include: Step S500: Obtain the measured time percentage and the baseline time percentage.

[0077] The proportion of actual time is consistent with the actual capacity factor in step S301, and is determined by the processing terminal through data extraction of the actual capacity factor.

[0078] The baseline time percentage is consistent with the baseline capacity factor in step S303, and is determined by the processing terminal through data extraction of the baseline capacity factor.

[0079] Step S501: Determine whether the overall offset index is greater than the preset overall offset threshold.

[0080] The overall shift threshold refers to the lower limit of the overall shift index corresponding to the occurrence of an overall shift in the chromatogram. If the overall shift index is greater than the overall shift threshold, it indicates that an overall shift has occurred in the chromatogram. The operator first determines multiple chromatographs that have been verified through pre-experiment and whose operating range under fixed elution parameters is within the baseline elution range. Then, the operator controls the chromatographs to elute the substance to be eluted multiple times to determine the normal overall shift index data corresponding to the fixed elution parameters. If the overall shift index is not normally distributed, the 99th quantile is taken as the overall shift threshold. If the overall shift index is normally distributed, the mean and standard deviation of the overall shift index are calculated. The overall shift threshold is obtained by subtracting three times the standard deviation from the mean of the overall shift index.

[0081] By processing the terminal to determine whether the overall offset index is greater than the overall offset threshold, it is determined whether the chromatogram has an overall offset. Then, based on the offset of the chromatogram, different compensation parameters are selected to compensate the mobile phase ratio parameters, thereby optimizing the operation of the chromatograph.

[0082] Step S5011: If it is greater than the specified value, analyze the proportion of the measured time and the proportion of the reference time to determine the fixed compensation parameters.

[0083] If the processing terminal determines whether the overall offset index is greater than the overall offset threshold, it indicates that an overall offset has occurred in the chromatogram. Therefore, the proportion of the measured time and the proportion of the reference time are analyzed to determine the fixed compensation parameter corresponding to the overall offset. The specific analysis steps are as follows: Figure 6 The steps in the process.

[0084] The fixed compensation parameter refers to the fixed compensation parameter corresponding to the mobile phase ratio of the chromatogram when there is an overall shift in the chromatogram relative to the reference elution data. It is consistent with the fixed compensation parameter in step S103 and is determined by the processing terminal through analysis of the proportion of the measured time and the proportion of the reference time. The specific analysis steps are as follows: Figure 6 The steps in the process.

[0085] Step S5012: If it is not greater than, then determine whether the partial offset index is greater than the preset partial offset threshold.

[0086] If the processing terminal determines that the overall offset index is not greater than the overall offset threshold, it indicates that the chromatogram has not shifted overall. Therefore, it is necessary to determine whether the partial offset index is greater than the partial offset threshold to determine whether the chromatogram has shifted partially relative to the reference elution data. Then, the corresponding fixed compensation parameters are determined based on the partial offset of the chromatogram.

[0087] The partial offset threshold refers to the lower limit of the partial offset threshold corresponding to a partial offset of the chromatogram relative to the reference elution data. If the partial offset index is greater than the overall offset threshold, it indicates that the chromatogram has partially shifted. The operator first determines multiple chromatographs that have been verified through pre-experimentation and whose operating range under fixed elution parameters is within the reference elution range. Then, the operator controls the chromatographs to elute the substance to be eluted multiple times to determine the normal partial offset index data corresponding to the fixed elution parameters. If the partial offset index is not normally distributed, the 99th quantile is taken as the overall offset threshold. If the overall offset index is normally distributed, the mean and standard deviation of the overall offset index are calculated. The overall offset threshold is obtained by adding three times the standard deviation to the mean of the overall offset index.

[0088] Step S5013: If it is greater than the reference time ratio, calculate the absolute deviation between the measured time ratio and the reference time ratio to determine the measured ratio deviation.

[0089] If the processing terminal determines that the partial offset index is greater than the partial offset threshold, it indicates that the chromatogram has partially shifted. Therefore, the measured percentage deviation is first calculated to provide data support for determining the fixed compensation parameters.

[0090] The measured proportion deviation refers to the absolute deviation between the measured time proportion and the reference time proportion, which is determined by the processing terminal by calculating the absolute deviation between the measured time proportion and the reference time proportion.

[0091] Step S5014: Analyze the measured percentage deviation, measured time percentage, and reference time percentage to determine the fixed compensation parameters.

[0092] The fixed compensation parameter refers to the fixed compensation parameter corresponding to the partial peak shift of the chromatogram relative to the reference elution data. It is consistent with the fixed compensation parameter in step S103 and is determined by the processing terminal through analysis of the measured proportion deviation, measured time proportion, and reference time proportion. The specific analysis steps are as follows: Figure 7 The steps in the process.

[0093] Step S5015: If it is not greater than, then the preset zero compensation parameter is determined as the fixed compensation parameter.

[0094] If the processing terminal determines that the partial offset index is not greater than the partial offset threshold, it indicates that the elution results of the chromatograph have experienced non-systematic drift due to environmental interference, edge fluctuations, etc., and there is no need to perform fixed compensation on the mobile phase ratio of the chromatograph. Therefore, the zero compensation parameter is determined as the fixed compensation parameter, and no fixed compensation is performed on the mobile phase ratio of the chromatograph. The elution process of the chromatograph is marked, and the operator checks the operation of the chromatograph to further determine the operation of the chromatograph and avoid misjudgment.

[0095] Zero compensation parameter refers to the fixed compensation parameter used when no adjustment is made to the fixed elution parameters of the chromatograph. The value of the zero compensation parameter is 0, and it is calibrated by the operator in the system.

[0096] Reference Figure 6 The steps for determining fixed compensation parameters by analyzing the proportion of measured time and the proportion of reference time include: Step S600: Calculate the relative deviation between the measured time percentage and the reference time percentage to determine the time percentage deviation.

[0097] Among them, the time proportion deviation refers to the relative deviation between the measured time proportion and the reference time proportion. It is determined by the processing terminal by first calculating the difference between the reference time proportion and the measured time proportion, and then calculating the quotient between the difference and the reference time proportion.

[0098] Step S601: Calculate the mean of the time percentage deviation to determine the mean of the percentage deviation.

[0099] The mean deviation of the proportion refers to the average value of the time proportion deviation of each effective elution component, which is determined by the processing terminal by calculating the average value of the time proportion deviation.

[0100] Step S602: Calculate the product of the average percentage deviation and the preset compensation adjustment factor to determine the initial overall compensation amount.

[0101] The compensation adjustment factor refers to the general adjustment factor used when adjusting the ratio of the organic phase in the mobile phase of the chromatograph to determine the reference value of the pre-experimental adjustment amount corresponding to the final fixed compensation factor. It is used to determine the reference point of the fine-tuning amount in subsequent fine-tuning experiments and improve the efficiency of the mobile phase ratio of the chromatograph. For different chromatographic separation modes such as reversed phase and ion exchange, different compensation adjustment factors are selected according to the ratio relationship corresponding to the mode. The operator determines the compensation adjustment factor by conducting offline calibration experiments on the chromatograph using different separation methods. First, the chromatograph is subjected to a fixed elution parameter experiment to determine that the elution parameters are normal and within the reference elution range. Then, a calibration experiment is set to determine the absolute value of the ratio of the relative deviation of the capacity factor of the corresponding effective elution component. The conversion coefficient corresponding to each separation method is then determined, which is the compensation adjustment factor.

[0102] The initial overall compensation amount refers to the initial reference value for fine-tuning the fixed compensation factor, which is determined by the compensation adjustment factor. It is determined by the processing terminal by calculating the product of the average percentage deviation and the compensation adjustment factor. Thus, when the chromatograph experiences an overall shift, the mobile phase ratio is adaptively adjusted based on the global average shift amount.

[0103] Step S603: Conduct a pre-elution experiment on the substance to be eluted based on the initial overall compensation amount and the preset fine-tuning upper limit to determine the fixed compensation parameters.

[0104] Among them, the upper limit of fine-tuning refers to the upper limit of the adjustment ratio for fine-tuning the mobile phase ratio in the fixed elution data, which is determined by the operator in the system according to the pharmacopoeia and legal data.

[0105] The fixed compensation parameter is consistent with the fixed compensation parameter in step S103. After determining the initial compensation parameter, the operator first truncates the initial compensation parameter according to the upper limit of the fine-tuning adjustment. When the initial compensation parameter is within the upper limit of the fine-tuning adjustment, the initial compensation parameter is used as the reference starting point. When the initial step size parameter is not within the fine-tuning adjustment range, a certain reserved redundancy value is set on the boundary of the fine-tuning adjustment range, and the fine-tuning amount after taking the redundancy value is used as the reference starting point. After determining the reference starting point, the reference starting point is expanded before and after according to the calibrated fine-tuning step size. Three sets of gradient fine-tuning experiments are set with the reference starting point as the center. In the gradient experiments, no less than 6 parallel injections are performed, and the fine-tuning amount is screened according to the baseline range requirements. The fine-tuning amount with the smallest adjustment amount that meets the baseline range requirements for the effective elution component is determined as the fixed compensation parameter. If there is no fine-tuning amount that meets the requirements, the gradient range is expanded for further screening. If a suitable fine-tuning parameter is still not determined, the operator is prompted to check the chromatograph according to the chromatograph fault information.

[0106] Reference Figure 7 The steps for determining fixed compensation parameters by analyzing the measured percentage deviation, measured time percentage, and reference time percentage include: Step S700: Perform numerical analysis on the measured percentage deviation to determine the maximum percentage deviation.

[0107] Among them, the maximum proportion deviation refers to the maximum value among the measured proportion deviations. The processing terminal determines the maximum value among the measured proportion deviations by traversing the measured proportion deviations, which is the maximum proportion deviation.

[0108] Step S701: Extract data on the baseline time percentage based on the maximum percentage deviation to determine the target baseline percentage.

[0109] Among them, the target reference ratio refers to the reference time ratio of the target adjustment reference corresponding to the fixed compensation parameter. After determining the maximum ratio deviation, the processing terminal determines the reference time ratio corresponding to the maximum ratio deviation as the target reference ratio.

[0110] Step S702: Calculate the quotient of the maximum percentage deviation and the target baseline percentage to determine the target relative deviation.

[0111] Among them, the target relative deviation refers to the relative deviation of the measured capacity factor that deviates the most from the reference capacity factor in the effective elution component. It is determined by the processing terminal by calculating the quotient of the maximum proportion deviation and the target reference proportion.

[0112] Step S703: Calculate the product of the target relative deviation and the preset compensation adjustment factor to determine the initial partial compensation amount.

[0113] The compensation adjustment factor is consistent with the compensation adjustment factor in step S602.

[0114] The initial partial compensation amount refers to the initial reference value of the fine-tuning amount used to determine the fixed compensation factor, which is determined by the compensation adjustment factor. It is determined by the processing terminal by calculating the product of the target relative deviation and the compensation adjustment factor, so that when the chromatograph has a partial deviation, the mobile phase ratio is adaptively adjusted according to the maximum target deviation.

[0115] Step S704: Conduct a pre-elution experiment on the substance to be eluted based on the initial partial compensation amount and the preset fine-tuning upper limit to determine the fixed compensation parameters.

[0116] The upper limit of the fine-tuning adjustment is consistent with the upper limit of the fine-tuning adjustment in step S603.

[0117] The fixed compensation parameter is consistent with the fixed compensation parameter in step S103. After determining the initial compensation parameter, the operator first truncates the initial compensation parameter according to the upper limit of the fine-tuning adjustment. When the initial compensation parameter is within the upper limit of the fine-tuning adjustment, the initial compensation parameter is used as the reference starting point. When the initial step size parameter is not within the fine-tuning adjustment range, a certain reserved redundancy value is set on the boundary of the fine-tuning adjustment range, and the fine-tuning amount after taking the redundancy value is used as the reference starting point. After determining the reference starting point, the reference starting point is expanded before and after according to the calibrated fine-tuning step size. Three sets of gradient fine-tuning experiments are set with the reference starting point as the center. In the gradient experiments, no less than 6 parallel injections are performed, and the fine-tuning amount is screened according to the baseline range requirements. The fine-tuning amount with the smallest adjustment amount that meets the baseline range requirements for the effective elution component is determined as the fixed compensation parameter. If there is no fine-tuning amount that meets the requirements, the gradient range is expanded for further screening. If a suitable fine-tuning parameter is still not determined, the operator is prompted to check the chromatograph according to the chromatograph fault information.

[0118] Based on the same inventive concept, embodiments of this application provide a chromatograph mobile phase adaptive mixing system, comprising: The acquisition module is used to acquire fixed elution parameters, baseline elution data, actual time percentage, and baseline time percentage. A memory for storing a program for an adaptive mixing method of the mobile phase in a chromatograph; The processor can load and execute programs in memory to implement an adaptive ratio method for the mobile phase of a chromatograph.

[0119] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the above-described division of functional modules is used as an example. In practical applications, the above functions can be assigned to different functional modules as needed, that is, the internal structure of the device can be divided into different functional modules to complete all or part of the functions described above. The specific working process of the system, device, and unit described above can be referred to the corresponding process in the foregoing method embodiments, and will not be repeated here.

[0120] The above are all preferred embodiments of this application and are not intended to limit the scope of protection of this application. Any feature disclosed in this specification (including the abstract and drawings) may be replaced by other equivalent or similar features unless specifically stated otherwise. That is, unless specifically stated otherwise, each feature is only one example of a series of equivalent or similar features.

Claims

1. A method for adaptive mobile phase ratio preparation in chromatographs, characterized in that, include: Obtain the preset fixed elution parameters and baseline elution data of the components to be separated; A preliminary elution experiment was conducted on the components to be separated using fixed elution parameters to determine the current elution data. The current elution data and the baseline elution data are analyzed to determine the overall offset index and the partial offset index; The overall offset index and partial offset index are analyzed to determine the fixed compensation parameters; The fixed elution parameters are compensated based on the fixed compensation parameters to determine the adaptive elution parameters.

2. The adaptive mobile phase mixing method for a chromatograph according to claim 1, characterized in that, The steps for analyzing current elution data and baseline elution data to determine the overall and partial shift indices include: Calculate the average value of the current elution data to determine the average measured data; Determine whether the average measured data meets the requirements of the baseline elution data; If so, the preset unbiased global index and the preset unbiased partial index are respectively determined as the global offset index and the partial offset index. If not, the average measured data and the baseline elution data are analyzed to determine the overall offset index and the partial offset index.

3. The adaptive mobile phase mixing method for a chromatograph according to claim 2, characterized in that, The steps for analyzing average measured data and baseline elution data to determine the overall and partial shift indices include: Data extraction was performed on the average measured data to determine the mean dead time, the mean chromatographic peak time, and the reference dead time; Calculate the relative proportion of the difference between the mean time value of the chromatographic peak and the mean dead time value to determine the measured capacity factor; Data extraction was performed on the baseline elution data to determine the baseline chromatographic time; Calculate the relative proportion of the difference between the baseline chromatographic time and the mean of the baseline dead time to determine the baseline capacity factor; The measured capacity factor and the baseline capacity factor are analyzed to determine the overall offset index and the partial offset index.

4. The adaptive mobile phase mixing method for a chromatograph according to claim 3, characterized in that, The steps for analyzing the measured capacity factor and the baseline capacity factor to determine the overall offset index and the partial offset index include: Calculate the quotient between the measured capacity factor and the baseline capacity factor to determine the measured shrinkage index; Numerical analysis of the measured contraction index was performed to determine the maximum and minimum contraction index. Calculate the quotient of the minimum contraction index and the maximum contraction index to determine the overall offset index; Calculate the absolute difference between the measured capacity factor and the reference capacity factor to determine the peak relative deviation; Numerical analysis of the relative peak deviation was performed to determine the maximum and minimum peak deviations. Calculate the difference between the maximum peak deviation and the minimum peak deviation to determine the partial offset index.

5. The adaptive mobile phase mixing method for a chromatograph according to claim 1, characterized in that, The steps for analyzing the overall and partial offset indices to determine the fixed compensation parameters include: Obtain the percentage of actual time and the percentage of baseline time; Determine whether the overall offset index is greater than the preset overall offset threshold; If it is greater than that, the proportion of measured time and the proportion of reference time will be analyzed to determine the fixed compensation parameters; If it is not greater than, then determine whether the partial offset index is greater than the preset partial offset threshold. If it is greater than the baseline time ratio, the absolute deviation between the measured time ratio and the baseline time ratio is calculated to determine the measured ratio deviation. The measured percentage deviation, measured time percentage, and reference time percentage are analyzed to determine the fixed compensation parameters; If it is not greater than, then the preset zero compensation parameter will be determined as the fixed compensation parameter.

6. The adaptive mobile phase mixing method for a chromatograph according to claim 5, characterized in that, The steps for analyzing the proportion of measured time and the proportion of baseline time to determine the fixed compensation parameters include: Calculate the relative deviation between the measured time percentage and the reference time percentage to determine the time percentage deviation; Calculate the mean of the time percentage deviation to determine the mean of the percentage deviation; Calculate the product of the average percentage deviation and the preset compensation adjustment factor to determine the initial overall compensation amount; Elution pre-experiments were conducted on the substances to be eluted based on the initial overall compensation amount and the preset fine-tuning upper limit to determine the fixed compensation parameters.

7. The adaptive mobile phase mixing method for a chromatograph according to claim 5, characterized in that, The steps for analyzing the measured percentage deviation, measured time percentage, and reference time percentage to determine the fixed compensation parameters include: Numerical analysis was performed on the measured percentage deviation to determine the maximum percentage deviation. Data on the baseline time percentage is extracted based on the maximum percentage deviation to determine the target baseline percentage; Calculate the quotient of the maximum percentage deviation and the target baseline percentage to determine the target relative deviation; Calculate the product of the target relative deviation and the preset compensation adjustment factor to determine the initial partial compensation amount; A preliminary elution experiment was conducted on the substance to be eluted based on the initial partial compensation amount and the preset upper limit for fine-tuning, in order to determine the fixed compensation parameters.

8. A mobile phase adaptive mixing system for a chromatograph, characterized in that, include: The acquisition module is used to acquire fixed elution parameters and baseline elution data; A memory for storing a program for an adaptive mobile phase ratio method for a chromatograph as described in any one of claims 1 to 7; The processor and the program in the memory can be loaded and executed by the processor to implement the adaptive ratio method of the mobile phase for chromatograph as described in any one of claims 1 to 7.

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