Methods for calculating the content of the tested target, detection system and sample analyzer

Abstract

This invention relates to a method for calculating the content of a analyte. The method includes: acquiring a complete chromatographic elution curve of the sample corresponding to the sample analyzer, returning to a baseline; detecting the chromatographic elution curve of the sample; based on the absorbance value at the end of the current sample measurement, extracting a segment from the complete chromatographic elution curve from the absorbance value at the end of the measurement to the baseline as a concentration-diffusion curve; correcting the chromatographic elution curve of the sample using the acquired concentration-diffusion curve; and calculating the content of the analyte based on the corrected chromatographic elution curve. This invention also relates to a detection system and sample analyzer using this calculation method. This invention can correct for the influence of sample residues on detection.

Description

Technical Field

[0001] This invention relates to a method for calculating the content of a target substance, a detection system using the calculation method, and a sample analyzer. Background Technology

[0002] Existing sample analyzers that utilize chromatography columns to detect target analytes generally require continuous and efficient separation and analysis of the target components in the sample. To improve detection speed, these analyzers typically employ high-concentration or highly polar mobile phases to elute the target analytes from the column more quickly. However, this results in a high-concentration component passing between the column and the detector. This residual solution remains in the tubing and connectors between the column and detector. When the residual concentration is high and the detection interval between two consecutive samples is short, the detector flow cell contains both residual sample solution from the previous sample and sample solution from the new sample, thus affecting the accuracy of the subsequent sample's detection results. Summary of the Invention

[0003] In view of the above, it is necessary to provide a method for calculating the content of the analyte that can correct for the influence of sample residues, a detection system using the method, and a sample analyzer.

[0004] A method for calculating the content of a analyte, applied to a sample analyzer, is disclosed to reduce the impact of residual samples on detection results during continuous testing. The sample analyzer performs chromatographic analysis on samples to determine the content of a specific analyte. The method for calculating the content of the analyte includes the following steps: obtaining a complete chromatographic elution curve of the sample corresponding to the sample analyzer, returning to a baseline; detecting the chromatographic elution curve of the sample; based on the absorbance value of the chromatographic elution curve of the sample at the end of the current measurement, extracting a segment from the obtained complete chromatographic elution curve of the sample from the absorbance at the end of the measurement to the baseline as a concentration-diffusion curve; using the obtained concentration-diffusion curve to correct the chromatographic elution curve of the next sample introduced at the end of the current measurement; and calculating the content of the analyte based on the chromatographic peak area corresponding to the analyte in the corrected chromatographic elution curve of the sample.

[0005] A detection system that calculates the content of a target substance by utilizing the peak area of ​​a chromatogram is applied to a sample analyzer to reduce the impact of residual samples on the detection results during continuous testing. The sample analyzer performs chromatographic analysis on the sample to determine the content of a specific analyte in the sample. The detection system includes: a reading module connected to the sample analyzer to read the chromatographic elution curve of the sample; a concentration diffusion curve acquisition module connected to the reading module to extract a segment of the complete chromatographic elution curve of the sample from the absorbance value at the end of the current measurement time point to the baseline, based on the absorbance value of the chromatographic elution curve of the sample at the end of the current measurement time point, as the concentration diffusion curve, wherein the complete chromatographic elution curve of the sample is formed by detecting all components in a single sample and returning to the baseline under the same detection conditions as the current measurement; a correction module connected to the reading module and the concentration diffusion curve acquisition module respectively to correct the chromatographic elution curve of the sample using the acquired concentration diffusion curve; and a calculation module connected to the correction module to calculate the content of the analyte based on the corrected detection data of the analyte.

[0006] A sample analysis method is provided for detecting the content of a specific analyte in a test sample. The sample analyzer includes a detection unit, a memory, and a processor. The detection unit performs chromatographic analysis on the test sample to obtain the chromatographic elution curve of the test sample. The memory stores the complete chromatographic elution curve of the test sample corresponding to the sample analyzer, returning to the baseline. The processor is connected to the detection unit and the memory. The processor extracts a segment from the complete chromatographic elution curve of the test sample, starting from the absorbance value at the end of the current sample measurement, up to the baseline, as a concentration-diffusion curve. This segment is used to correct the chromatographic elution curve of the next sample introduced at the end of the current measurement. The content of the analyte is calculated based on the chromatographic peak area corresponding to the analyte in the corrected chromatographic elution curve of the test sample.

[0007] Compared with the prior art, the method for calculating the content of the target substance provided by the present invention, the detection system using the method, and the sample analyzer correct the residual effect of the sample when the next sample is detected without returning to the baseline during continuous detection by measuring or simulating the complete chromatographic elution curve of a specific sample under preset detection conditions. This improves the efficiency of sample analysis while ensuring the accuracy of detection. Attached Figure Description

[0008] Figure 1 This is a schematic diagram of the sample analyzer and its detection system provided in an embodiment of the present invention.

[0009] Figure 2 For use Figure 1 A schematic diagram of the complete chromatographic elution curve obtained by the sample analyzer.

[0010] Figure 3 To adopt in Figure 2 A schematic diagram of the initial segment of the chromatographic elution curve obtained by the concentration diffusion curve correction sample analyzer.

[0011] Figure 4 To adopt in Figure 2 The obtained concentration diffusion curve is a schematic diagram of the end segment of the chromatographic elution curve measured by the supplementary sample analyzer.

[0012] Figure 5 A flowchart illustrating the steps of the data processing method provided in this embodiment of the invention.

[0013] Figure 6 To adopt in Figure 2 A schematic diagram of the initial segment of the chromatographic elution curve of the (x+1)th sample obtained by the concentration diffusion curve correction sample analyzer.

[0014] Figure 7 To adopt in Figure 2 The obtained concentration diffusion curve is a schematic diagram of the end segment of the chromatographic elution curve of the (x+1)th sample measured by the supplementary sample analyzer. Detailed Implementation

[0015] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0016] like Figure 1As shown, the sample analyzer 1 provided in this embodiment of the invention is used for semi-quantitative / quantitative determination of specific analytes in a sample 2. The sample analyzer 1 includes a sample unit 10, a reagent unit 11, a mixing unit 12, an elution unit 13, a loading unit 14, a separation unit 15, a detection unit 16, a memory 17, and a processor 18. The sample unit 10 provides the sample 2. The reagent unit 11 provides the reagents required for detection. The reagent unit 11 and the sample unit 10 are respectively connected to the mixing unit 12 to mix the sample and reagents into a sample solution. The mixing unit 12 is connected to the separation unit 15 via the loading unit 14 to load the sample solution into the separation unit 15 for component separation. The elution unit 13 is connected to the separation unit 15 via the loading unit 14 to provide eluent to the separation unit 15. The separation unit 15 separates the analytes from the sample solution using the eluent. The detection unit 16 is connected to the separation unit 15 to perform chromatographic analysis on the analytes separated by the separation unit 15. The sample unit 10, reagent unit 11, mixing unit 12, elution unit 13, loading unit 14, memory 17 and processor 18 are electrically connected directly or indirectly via a bus to transmit and exchange data or signals, thereby achieving automated control.

[0017] In this embodiment, the sample analyzer 1 is a glycated hemoglobin analyzer, which uses high-performance liquid chromatography (HPLC) to detect the content of glycated hemoglobin (HbA1c) in a blood sample. Correspondingly, the analyte is HbA1c. The sample analyzer 1 separates HbA1c and other hemoglobin components, mainly hemoglobin A0 (HbA0), from the blood sample through the separation unit 15, and then detects the HbA1c content. It is understood that in other embodiments, the analyte can also be various protein components with different charges.

[0018] In addition, depending on the characteristics of the sample being tested, the sample analyzer 1 can also use other detection methods, such as gas chromatography analysis, and is not limited to this embodiment.

[0019] The memory 17 stores an operating system for controlling the operation of the sample analyzer 1. The operating system is run by the processor 18. The sample analyzer 1 performs various tests under the control and coordination of the operating system.

[0020] The sample unit 10 includes a sample tank 100 and a sample pump 102. The sample tank 100 contains the sample to be tested 2. The sample tank 100 is connected to a mixing unit 12 via a conduit to form a liquid path for transferring the sample to be tested 2 to the mixing unit 12. The sample pump 102 is disposed on the liquid path between the sample tank 100 and the mixing unit 12 to draw samples into the mixing unit 12 at a set flow rate. In this embodiment, the sample to be tested 2 is a blood sample.

[0021] The reagent unit 11 includes a reagent tank 110 and a reagent pump 112. The reagent tank 110 contains reagents for pre-reaction with the sample 2 before testing. The reagent tank 110 is connected to a mixing unit 12 via a conduit to form a liquid path for transferring reagents to the mixing unit 12. The reagent pump 112 is located on the liquid path between the reagent tank 110 and the mixing unit 12 to draw reagents into the mixing unit 12 at a set flow rate. It is understood that multiple reagents can be configured according to actual needs, each reagent being contained in a separate reagent tank 110 and delivered to the mixing unit 12 by a corresponding reagent pump 112. In this embodiment, the reagents include a hemolytic agent and / or a buffer.

[0022] The mixing unit 12 is used to hold the introduced test sample 2 and reagents. The test sample 2 and reagents are uniformly mixed in the mixing unit 12 for a pre-reaction before detection. For example, in the detection of glycated hemoglobin, the blood sample to be tested must first be mixed with a hemolysis reagent to perform a hemolysis reaction in order to release hemoglobin bound to blood glucose from blood cells. The mixed test sample 2 and reagents are loaded into the separation unit 15 through the loading unit 14 for component separation.

[0023] It is understandable that the liquid path between each unit can also be established by using a pipette to draw liquid, move it to the destination, and then release the liquid to transfer liquid between units, in order to replace the liquid path transfer method of connecting pipes in this embodiment.

[0024] The separation unit 15 is used to separate the analyte from the sample solution and bring it to the detection unit 16 for detection. The separation unit 15 can be a chromatography column or other components capable of separating material components. One end of the separation unit 15 is connected to the loading unit 14 via a conduit to receive a specific loading volume of sample solution. The separation unit 15 contains a packing material. The packing material selectively adsorbs specific substances in the sample solution to be tested, thereby separating the analyte from the sample solution.

[0025] The elution unit 13 loads the eluent into the separation unit 15 through the loading unit 14 to elute the specific substances that have been separated.

[0026] In this embodiment, the sample analyzer 1 is used to determine the content of HbA1c in blood, and the filler is a substance that can selectively adsorb HbA1c in the sample solution to be tested, such as methacrylate copolymer.

[0027] The detection unit 16 includes a chromatographic detector 160 and a flow cell 162. One end of the flow cell 162 is connected to the end of the separation unit 15 furthest from the loading unit 14. Figure 2 As shown, the mobile phase flowing out after separation by separation unit 15 flows through flow cell 162. The chromatographic detector 160 performs chromatographic analysis on the mobile phase flowing through flow cell 162 to detect the mobile phase at a preset time point. T The absorbance was measured to obtain the chromatographic elution curve for a certain time range, with time on the horizontal axis and absorbance on the vertical axis. f ( A , T ).

[0028] According to Beer-Lambert's law, when a beam of monochromatic light shines on the surface of an absorbing medium and passes through a medium of a certain thickness, the intensity of the transmitted light decreases because the medium absorbs some of the light energy. The higher the concentration and the thicker the absorbing medium, the more significant the decrease in light intensity. The relationship is shown in Equation 1:

[0029] A = = = Formula 1

[0030] in A Absorbance I 0 The intensity of the incident light. I t The intensity of the transmitted light. T For light transmittance, K It is a coefficient (usually the molar absorption coefficient). ε ), l The thickness of the absorbing medium, c This represents the concentration of the light-absorbing substance.

[0031] Formula 1 shows that when a beam of parallel monochromatic light passes perpendicularly through a uniform, non-scattering absorbing material, the absorbance is... A With the concentration of light-absorbing substances c and the thickness of the absorption layer l The absorbance is directly proportional to the absorbance of each component. When there are multiple absorbing components in a medium, the absorbance is additive, provided that the components do not interact with each other. That is to say, the total absorbance of the medium at a certain wavelength is the sum of the absorbances of each component at that wavelength.

[0032] For sample analyzer 1 in this embodiment,l The absorbance detected by detector 50 is a fixed value. A The concentration of substances with luminescent groups in the eluent c Proportional. The chromatographic elution curve obtained by the sample analyzer 1 f ( A , T The various chromatographic peaks correspond to the absorbant components eluted from the separation unit 15 of the sample 2 at different times. The peak area of ​​each chromatographic peak, i.e., the absorbance... A During the time period Δ corresponding to the chromatographic peak t The integral within the sample is proportional to the concentration of each absorbant component in the sample 2 being tested. This is calculated using the molar absorptivity preset in the sample analyzer 1. ε Or the standard curves of each absorbance component versus the resulting chromatographic elution curves f ( A , T By fitting and calculating the peak area of ​​each chromatographic peak, the content and proportion of the analyte in the sample 2 can be obtained.

[0033] However, when sample residue exists in the tubing and connectors between the separation unit 15 and the flow cell 162 of the detection unit 16, the flow cell 162 of the detection unit 16 contains not only the eluent of the current sample 2 but also the residual eluent of the previous sample 2. Therefore, based on the additive nature of absorbance, the absorbance of the analyte detected by the detection unit 16 will be... A1 It is not the actual absorbance of the analyte in the sample. A It also includes the absorbance of the test sample remaining in the previous sample 2. A2 Therefore, when sample residue is present, the detected chromatographic elution curve contains errors, affecting the analysis results of sample 2. It is understood that the "sample residue phenomenon" includes, but is not limited to, the situation where the previous sample remains in the tubing and connectors between the flow cell 162 of the separation unit 15 and the detection unit 16, as well as the situation where the previous sample remains in other tubing and connectors outside of the separation unit 15 and the detection unit 16 of the sample analyzer 1.

[0034] The present invention also provides a method for applying chromatographic elution curves to the sample analyzer 1. f ( A , T The detection system 19, which calculates the content of the target substance by the peak area, is used to correct the detection data of the sample analyzer 1 affected by residual samples. The detection system 19 can be a subsystem of the operating system of the sample analyzer 1, or it can be stored in the memory 17 independently of the operating system, or it can be firmware solidified on the memory 17 and not directly controlled by the operating system.

[0035] The detection system 19 includes a reading module 190, a concentration diffusion curve acquisition module 192, a correction module 194, a storage module 196, and a calculation module 198. The reading module 190 is connected to the memory 17 and / or the detection unit 16 to read the detection data of the analyte measured by the detection unit 16. The concentration diffusion curve acquisition module 192 is connected to the detection unit 16 and / or the memory 17 to acquire a concentration diffusion curve used to correct the detection data. f d ( A , T The correction module 194 is connected to the concentration diffusion curve acquisition module 192 and the reading module 190, so as to obtain the concentration diffusion curve. f d ( A , T The detection data of the target substance being tested is corrected. The storage module 196 is connected to the correction module 194 and the memory 17, and is used to store the corrected detection data in the memory 17. The calculation module 198 is connected to the correction module 194 and / or the storage module 196 to calculate the concentration or content ratio of the target substance based on the corrected detection data. The modules 190-198 can be a set of logical relationships embedded in hardware or firmware, or a series of programs written in a programming language and stored in the memory 17 or other firmware. The programs can be executed by at least one of the processors 18 to implement specific functions.

[0036] The reading module 190 can directly read the detection data of the target object from the detection unit 16, or it can read the detection data of the target object already stored in the memory 17. The detection data of the target object read by the reading module 190 can be directly sent to the correction module 194 for processing, or it can be cached for later processing.

[0037] like Figure 2 As shown, the concentration diffusion curve f d ( A , T () is defined as the complete chromatographic elution curve of a specific sample. f ( A , T The curve, starting from a preset time point and continuing until it returns to the baseline, reflects the concentration of residues in the sample solution eluted from separation unit 15 during detection, as the residue diffuses away over time. This concentration diffusion curve... f d ( A , TThe complete chromatographic elution curve extracted f ( A , T The paragraph is based on the chromatographic elution curve measured at the end of the measurement. f ' ( A , T It depends on the situation at baseline 3. Because of the concentration diffusion curve... f d ( A , T The concentration diffusion curve is typically used to correct for the influence of residues in the eluted sample solution near the end of the previous sample's testing on subsequent sample testing. f d ( A , T () is the complete chromatographic elution curve for single-sample detection. f ( A , T The section near the end. The complete chromatographic elution curve. f ( A , T A complete chromatographic elution curve is defined as the absorbance profile of a single sample after all components have been eluted and eluted from the flow cell. f ( A , T The absorbance at the end time point is the same as the absorbance at the start time point, indicating a complete chromatographic elution curve. f ( A , T Returning to baseline 3. If a zeroing operation is performed before the detection begins, then the complete chromatographic elution curve will be obtained. f ( A , T The absorbance at the start and end times should both be zero.

[0038] The complete chromatographic elution curve f ( A , T The shape of the concentration diffusion curve is affected by detection conditions, such as the hardware settings and detection parameters of the sample analyzer 1. These detection parameters include, but are not limited to, flow rate, eluent composition, and elution gradient. Therefore, the concentration diffusion curve... f d ( A , T It is necessary to make special settings for specific sample analyzers 1 to use the same detection conditions to detect specific samples.

[0039] The complete chromatographic elution curve of the sample analyzer 1 under preset detection conditions f( A , T The chromatographic elution curve can be preset and stored in memory 17 as a reference standard, and can be used in actual detection based on the measured chromatographic elution curve. f ' ( A , T The absorbance at the end of the measurement is obtained from the complete chromatographic elution curve. f ( A , T The curve segment from the absorbance at the end of the measurement to the baseline is taken as the concentration-diffusion curve corresponding to this detection. f d ( A , T (For use in subsequent revisions)

[0040] The complete chromatographic elution curve f ( A , T The concentration diffusion curve acquisition module 192 can simulate the complete chromatographic elution curve in real time based on the user's current detection conditions and preset sample solution diffusion rules. f ( A , T The complete chromatographic elution curve can be stored in memory 17 for later retrieval. f ( A , T It can also be obtained by recording the chromatographic elution curve of the last added sample after setting the detection conditions. The non-continuous sample addition method refers to testing a single sample or stopping sample addition after continuous sample addition testing, and then recording the complete chromatographic elution curve of the last added sample. f ( A , T ).

[0041] In this embodiment, the sample analyzer needs to detect the HbA1c content in the blood sample. For example... Figure 2 As shown, the concentration diffusion curve f d ( A , T ( ) represents the complete chromatographic elution curve. f ( A , T The curve between the time point at the end of the HbA0 chromatographic peak and the time point at which the absorbance returns to baseline 3 in the final segment, i.e. Figure 2The portion to the right of the dashed line. The measurement end point can be calculated based on the tubing volume of the sample analyzer 1 and the set flow rate. In continuous injection detection mode, this measurement end point is the detection start point for the next sample. Ideally, the chromatographic curve at this measurement end point should return to baseline 3. If the chromatographic curve at this measurement end point does not return to baseline 3, then the concentration diffusion curve... f d ( A , T This reflects the diffusion process of HbA0 concentration remaining in the pipeline and connectors under the set detection conditions of this sample analyzer 1.

[0042] The correction module 194 checks the chromatographic elution curve of the measured sample solution. f ' ( A , T To determine whether correction is needed, if the chromatographic elution curve... f ' ( A , T If correction is needed, the complete chromatographic elution curve obtained from module 192 of the concentration diffusion curve acquisition module will be used. f ( A , T Extract the corresponding concentration diffusion curve from the data. f d ( A , T The measured chromatographic elution curve f ' ( A , T Corrections are made to the chromatographic elution curve. f ' ( A , T No correction is needed for the chromatographic elution curve. f ' ( A , T The correction module 194 performs data analysis or output. It then determines the measured chromatographic elution curve. f ' ( A , T The criterion for whether correction is needed is the measured chromatographic elution curve. f ' ( A , T ) At the end of the measurement, has the chromatographic elution curve returned to baseline 3? f ' ( A, T The chromatographic elution curve did not regress to baseline 3. f ' ( A , T If the absorbance at the end of the measurement is higher than the absorbance at the beginning of the measurement, then the measured chromatographic elution curve is considered to be accurate. f ' ( A , T (This needs correction if the measured chromatographic elution curve is...) f ' ( A , T If the absorbance has returned to baseline 3, meaning the absorbance at the end of the measurement is equal to the absorbance at the beginning of the measurement, then the measured chromatographic elution curve is considered to have returned to baseline 3. f ' ( A , T No correction is needed.

[0043] When the measured chromatographic elution curve f ' ( A , T When correction is needed, the correction module 194 acquires the measured chromatographic elution curve. f ' ( A , T The absorbance value at the end of the measurement, and the complete chromatographic elution curve. f ( A , T Find the chromatographic elution curve that matches the measured value. f ' ( A , T The complete chromatographic elution curve is obtained by using the correction starting point corresponding to the absorbance value at the end of the measurement. f ( A , T The section from the starting point of this correction until the regression to baseline 3 is used as the concentration diffusion curve. f d ( A , T The measured chromatographic elution curve f ' ( A , T Corrections are made based on the concentration of the analyte in different samples during the actual testing process. This results in different absorbance values ​​measured at the end of the measurement. Therefore, the concentration-diffusion curve corresponding to the measured absorbance value is used for correction. f d (A , T The results will also be different.

[0044] In other embodiments, the correction module 194 can also convert the complete chromatographic elution curve according to the time interval between the measurement end point and the measurement start point. f ( A , T The point on the curve whose distance from the starting point is equal to the time interval is determined as the correction starting point, and then the complete chromatographic elution curve is plotted. f ( A , T The section from the starting point of the correction until the regression to the baseline is used as the concentration diffusion curve. f d ( A , T The measured chromatographic elution curve is then corrected.

[0045] like Figure 3 As shown, the correction module 194 can utilize the concentration diffusion curve. f d ( A , T ) The chromatographic elution curve of the next sample introduced at the end of this measurement. f ' ( A , T Corrections were made. This was because the chromatographic elution curve was corrected at the end of the measurement for the current sample. f ' ( A , T Since the absorbance of the sample has not yet returned to baseline 3, according to the additivity of absorbance, the absorbance of a specific component measured by the sample analyzer 1 at a certain time point when measuring the next sample is equal to the sum of the actual absorbance of that specific component in the measured sample and the absorbance of the residue in the previous sample. Therefore, in order to obtain the actual absorbance of that specific component in the current sample, it is necessary to subtract the absorbance of the residue in the previous sample from the absorbance of that specific component detected at that time point, as shown in Formula 2:

[0046] Formula 2

[0047] in, A’ This represents the actual absorbance of a specific component in the currently tested sample. A * The absorbance measured by sample analyzer 1 A d The absorbance of residues in the tubing or connectors of sample analyzer 1. ε' The molar absorption coefficient of this specific component. εd The molar absorption coefficient of the residue. l The optical path length of the pipeline. c’ This represents the actual concentration of the specific component in the sample being tested. c d This represents the concentration of residue. As shown in the figure, for samples with residue requiring chromatographic elution, the elution curve is... f ' ( A , T When making corrections, sample analyzer 1 will display the measured chromatographic elution curve. f ' ( A , T The concentration diffusion curve of the previous sample remaining within the same time period is subtracted from the starting time point. f d ( A , T This allows us to obtain the actual chromatographic elution curve of the sample being tested. f ‘ r ( A , T This allows for the correction of sample analysis results.

[0048] like Figure 4 As shown, the correction module 194 can also utilize the concentration diffusion curve. f d ( A , T The measured chromatographic elution curve f ' ( A , T The portion from the end of the measurement to the regression baseline is supplemented and completed.

[0049] The storage module 196 stores the corrected chromatographic elution curve. f ‘ r ( A , T Save the corrected chromatographic elution curve. f ‘ r ( A , T The data can be stored in the memory 17 or cached for later processing.

[0050] The calculation module 198 calculates the chromatographic elution curve of the tested sample based on the corrected chromatographic elution curve. f ‘ r ( A ,T The concentration of the analyte is calculated from the chromatographic peak area corresponding to the analyte in the calculation module 198. The calculation module 198 can also supplement the actual chromatographic elution curve of the analyte sample with the correction module 194. f ‘ r ( A , T The ratio of the total area of ​​the test sample or the area of ​​other chromatographic peaks to the area of ​​the chromatographic peak corresponding to the test sample is used to calculate the content ratio of the test sample to the content of other components in the test sample.

[0051] The present invention will be further described below with reference to the data processing method of the detection system 19 applied to the above-mentioned sample analyzer 1.

[0052] like Figure 5 As shown, Figure 5 A flowchart of a method for calculating the content of a target substance applied to the detection system 19 of the sample analyzer 1, provided by an embodiment of the present invention, wherein the data processing method includes the following steps:

[0053] Step S100: Obtain the complete chromatographic elution curve corresponding to the preset detection conditions. f ( A , T The complete chromatographic elution curve f ( A , T The chromatographic elution curve (CEC) measured by sample analyzer 1 under preset detection conditions is defined as the complete chromatographic elution curve that can revert to baseline 3. f ( A , T ).

[0054] The complete chromatographic elution curve f ( A , T The complete chromatographic elution curve can be preset and stored in memory 17. f ( A , T It can also be simulated in real time based on the user's current detection conditions and preset sample solution diffusion patterns, resulting in a complete simulated chromatographic elution curve. f ( A , T It can be stored in memory 17 for later retrieval.

[0055] It is understood that, in other embodiments, the concentration diffusion curve acquisition module 192 can also record the complete chromatographic elution curve of the sample under the set detection conditions by means of discontinuous sample addition, provided that the detection conditions are set. f (A , T The non-continuous sample addition method refers to testing a single sample or stopping sample addition after continuous sample addition testing, and testing the complete chromatographic elution curve of the last added sample. f ( A , T ).

[0056] Step S101: Read the chromatographic elution curve of the sample being tested. f ' ( A , T The data can be read directly from the detection unit 16, or it can read the detection data of the sample already stored in the memory 17.

[0057] Step S102: Determine whether it is necessary to analyze the measured chromatographic elution curve. f ' ( A , T Corrections are made based on the measured chromatographic elution curve. f ' ( A , T The degree to which the measured chromatographic elution curve reverts to baseline 3 at the end of the measurement is used to determine whether further analysis of the measured elution curve is necessary. f ' ( A , T Modify the chromatographic elution curve as needed. f ' ( A , T If the absorbance at the end of the measurement is higher than that at the beginning of the measurement, then the measured chromatographic elution curve is considered to have failed to return to baseline 3. f ' ( A , T (This needs correction if the measured chromatographic elution curve is...) f ' ( A , T If the absorbance has returned to baseline 3, meaning the absorbance at the end of the measurement is equal to the absorbance at the beginning of the measurement, then the measured chromatographic elution curve is considered to have returned to baseline 3. f ' ( A , T No correction is needed.

[0058] Step S103, based on the measured chromatographic elution curve f ' ( A , TThe extent to which baseline 3 regresses from the complete chromatographic elution curve at the end of the measurement time point. f ( A , T Extract the corresponding concentration diffusion curve from the data. f d ( A , T ).

[0059] Specifically, the measured chromatographic elution curve can be obtained first. f ' ( A , T ) Measure the absorbance value at the end time point and include it in the complete chromatographic elution curve. f ( A , T Find the chromatographic elution curve that matches the measured value. f ' ( A , T The corrected starting point is the same as the absorbance value at the end of the measurement, and the complete chromatographic elution curve is obtained. f ( A , T The section from the starting point of this correction until the regression to baseline 3 is used as the concentration diffusion curve. f d ( A , T The measured chromatographic elution curve f ' ( A , T (This needs to be corrected.)

[0060] In other embodiments, the measured chromatographic elution curve can also be used as a reference. f ' ( A , T The time interval between the end and start points of the measurement will complete the chromatographic elution profile. f ( A , T The point on the curve whose distance from the starting point is equal to the time interval is determined as the correction starting point, and then the complete chromatographic elution curve is plotted. f ( A , T The section from the starting point of this correction until the regression to baseline 3 is used as the concentration diffusion curve. f d ( A , T The measured chromatographic elution curve f ' ( A ,T (This needs to be corrected.)

[0061] Step S104, based on the obtained concentration diffusion curve f d ( A , T The measured chromatographic elution curve f ' ( A , T (This needs to be corrected.)

[0062] The concentration diffusion curve f d ( A , T This can be used to generate the chromatographic elution profile for the next sample introduced at the end of this measurement. f ' ( A , T Corrections are made based on the chromatographic elution curve measured at the end of the measurement for the current sample. f ' ( A , T Since the absorbance of the sample has not yet returned to baseline 3, according to the additivity of absorbance, the absorbance of a specific component measured by the sample analyzer 1 at a certain time point when measuring the next sample is equal to the sum of the actual absorbance of that specific component in the measured sample and the absorbance of the residue in the previous sample. Therefore, in order to obtain the actual absorbance of that specific component in the current sample, it is necessary to subtract the absorbance of the residue in the previous sample from the absorbance of that specific component detected at that time point.

[0063] Furthermore, the concentration diffusion curve f d ( A , T It can also be used to complete the measured chromatographic elution curve from the end of the measurement to the regression baseline.

[0064] For example, when sample analyzer 1 continuously tests n When there are 1 sample, the detection time for each sample is 1. t If the time is seconds, then the chromatographic elution curve for each sample is expressed as follows: f 1( A , T ), f 2( A , T ), ……, f x ( A , T ), f x+1 (A , T ), ……, f n (A, T ), where the chromatographic elution curve of the x-th sample. f x ( A , T The measurement start time point is t x The test ends at ( t x + t ).

[0065] like Figure 6 As shown, for the initial segment of the chromatographic elution curve of the (x+1)th sample, if the starting time point of the xth sample test... t x absorbance value A [ t x Less than the test end time ( t x + t The absorbance value at that time A [ t x + t This indicates that a certain concentration of the sample remained in the tubing or connector of analyzer 1 at the end of the x-th sample test. To eliminate the influence of sample residue, the complete chromatographic elution curve was analyzed. f ( A , T Select from ) A [ t x + t [Numerical starting point until the complete chromatographic elution curve] f ( A , T The segment regressing to baseline 3 is used as the concentration diffusion curve. f d ( A [ t x + t The chromatographic elution curve of the (x+1)th sample. f x+1 ( A , T From the starting point of the time period t x+1 Start subtracting f d ( A [ t x +t The corrected chromatographic elution curve of the sample was obtained. .

[0066] like Figure 7 As shown, for the ending segment of the chromatographic elution curve of the (x+1)th sample, if the starting time point of the (x+1)th sample test... t x+1 absorbance value at time A [ t x+1 Less than the test end time ( t x+1 + t absorbance value at time ) A [ t x+1 + t When the test for the (x+1)th sample ends, it indicates that a certain concentration of the tested sample still remains in the tubing or connector of the sample analyzer 1. To obtain the complete chromatographic elution curve for the (x+1)th sample, the complete chromatographic elution curve is obtained from... f ( A , T Select from ) A [ t x+1 + t The range from the initial value to the baseline is used as the concentration diffusion curve. f d ( A [ t x+1 + t ]).Will f d ( A [ t x+1 + t The curve is supplemented by the chromatographic elution curve of this sample. f x+1 ( A , T After that, that is f x+1 ( A , T The value is extended to the position of the regression baseline. Figure 7 The portion to the left of the dashed line shows the chromatographic elution curve at the end of the test, while the portion to the right of the dashed line shows the selected characteristic curve. f d ( A [ t x+1 + t ]).

[0067] The corrected chromatographic elution curve of the (x+1)th sample It can more accurately reflect the concentration characteristics of each component in the sample, thereby accurately calculating the content and proportion of the target substance in the sample.

[0068] Step S105, the corrected chromatographic elution curve is obtained. f ‘ r ( A , T Save the corrected chromatographic elution curve. f ‘ r ( A , T The data can be stored in the memory 17 or cached for later processing.

[0069] Step S106: Calculate the concentration or content ratio of the analyte, and then use the corrected chromatographic elution curve of the analyte sample. f ‘ r ( A , T The concentration of the analyte can be calculated from the chromatographic peak area corresponding to the analyte in the sample. Additionally, the concentration can be determined based on the complete chromatographic elution curve of the sample. f ‘ r ( A , T The ratio of the total area of ​​the chromatographic peak of the analyte to the area of ​​the chromatographic peak of the analyte, or the ratio of the area of ​​the chromatographic peak of the analyte to the area of ​​the chromatographic peak of the analyte, is used to calculate the content ratio of the analyte to the sample or other components in the sample.

[0070] The detection system of the sample analyzer 1 and the data processing method applied to the detection system correct the residual effect of samples that are not returned to the baseline when the next sample is detected during continuous detection by measuring or simulating the complete chromatographic elution curve of a specific sample returning to the baseline under preset detection conditions. This improves the efficiency of sample analysis while ensuring the accuracy of detection.

Claims

1. A method for calculating the content of a analyte, applied to a sample analyzer to reduce the impact of residual samples on the detection results during continuous testing, wherein the sample analyzer performs chromatographic analysis on the sample to determine the content of the analyte in the sample, characterized in that, The method for calculating the content of the tested standard includes the following steps: Obtain the complete chromatographic elution curve of the test sample corresponding to the sample analyzer, which is regressed to the baseline. Detect the chromatographic elution curve of the sample being tested; Based on the absorbance value at the end of the measurement of the current sample, a segment of the complete chromatographic elution curve of the sample is extracted from the absorbance value at the end of the measurement until the return to the baseline, and is taken as the concentration diffusion curve. The obtained concentration diffusion curve is used to correct the chromatographic elution curve of the measured sample; wherein, based on the concentration diffusion curve, the chromatographic elution curve of the measured sample is supplemented from the measurement end point to the return baseline, or the absorbance value of the chromatographic elution curve of the measured sample is subtracted from the absorbance of the sample residue based on the concentration diffusion curve, so as to correct the chromatographic elution curve of the measured sample. The content of the analyte is calculated based on the chromatographic peak area corresponding to the analyte in the chromatographic elution curve of the corrected sample.

2. The method for calculating the content of the tested standard as described in claim 1, characterized in that: Before obtaining the concentration diffusion curve, determine whether the chromatographic elution curve of the sample has returned to the baseline. If it has not returned to the baseline, obtain the concentration diffusion curve to correct the chromatographic elution curve of the sample.

3. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The complete chromatographic elution curve is obtained by simulating the detection conditions of the sample analyzer and the preset sample diffusion law.

4. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The complete chromatographic elution curve is obtained by recording the chromatographic elution curve of the sample after all components have been detected and the sample has returned to the baseline using a sample analyzer in a non-continuous sample addition manner.

5. The method for calculating the content of the tested standard as described in claim 4, characterized in that: The non-continuous sample addition method includes testing a single sample or stopping sample addition after continuous sample addition testing and testing the complete chromatographic elution curve of the last added sample.

6. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The chromatographic elution curve of the next sample is corrected by subtracting the concentration diffusion curve from the chromatographic elution curve of the next sample measured at the current measurement end time point.

7. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The chromatographic elution curve of the current sample is corrected by using the concentration diffusion curve to supplement the portion of the chromatographic elution curve of the current sample from the end of the measurement to the return baseline.

8. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The ratio of the content of the target substance to the content of the target substance or other components in the test sample is calculated based on the ratio of the total area of ​​the chromatographic elution curve of the completed test sample or the chromatographic peak area of ​​other components to the chromatographic peak area corresponding to the target substance.

9. The method for calculating the content of the tested standard as described in claim 1, characterized in that: The concentration of the analyte is calculated based on the peak area corresponding to the analyte in the chromatographic elution curve of the corrected sample.

10. A detection system for calculating the content of a analyte using chromatographic peak area, applied to a sample analyzer to reduce the impact of residual samples on detection results during continuous testing, wherein the sample analyzer performs chromatographic analysis on samples to determine the content of the analyte in the samples, the detection system comprising: A reading module is connected to the sample analyzer to read the chromatographic elution curve of the sample being tested; The concentration diffusion curve acquisition module is connected to the reading module to extract a segment of the complete chromatographic elution curve of the sample from the absorbance value at the current measurement end time point to the baseline, based on the absorbance value of the chromatographic elution curve of the sample. The complete chromatographic elution curve of the sample is the chromatographic elution curve formed when all components in a single sample are detected and returned to the baseline under the same detection conditions as the current one. The correction module is connected to both the reading module and the concentration diffusion curve acquisition module to correct the chromatographic elution curve of the measured sample using the acquired concentration diffusion curve. Specifically, based on the concentration diffusion curve, the chromatographic elution curve of the measured sample is supplemented from the measurement end point to the return baseline, or the absorbance value of the chromatographic elution curve of the measured sample is subtracted from the absorbance of the sample residue based on the concentration diffusion curve to correct the chromatographic elution curve of the measured sample. and The calculation module is connected to the correction module to calculate the content of the test substance based on the corrected detection data of the test substance.

11. The detection system as described in claim 10, characterized in that, The complete chromatographic elution curve is obtained by simulating the detection conditions of the sample analyzer and the preset sample diffusion law.

12. The detection system as described in claim 10, characterized in that, The complete chromatographic elution curve is obtained by recording the chromatographic elution curve of the sample after all components have been detected and the sample has returned to the baseline using a sample analyzer in a non-continuous sample addition manner.

13. The detection system as described in claim 12, characterized in that, The non-continuous sample addition method includes testing a single sample or stopping sample addition after continuous sample addition testing and testing the complete chromatographic elution curve of the last added sample.

14. The detection system as described in claim 10, characterized in that, The correction module corrects the chromatographic elution curve of the next sample measured at the current measurement end time point by subtracting the concentration diffusion curve from the starting time point.

15. The detection system as described in claim 10, characterized in that, The correction module corrects the chromatographic elution curve of the current sample by using the concentration diffusion curve to supplement and complete the chromatographic elution curve of the current sample from the measurement end point to the return baseline.

16. A sample analyzer for detecting the content of a analyte in a sample, the sample analyzer comprising a detection unit, a memory, and a processor, wherein the detection unit performs chromatographic analysis on the sample to obtain a chromatographic elution curve of the sample, the memory stores a complete chromatographic elution curve of the sample corresponding to the sample analyzer, the processor is connected to the detection unit and the memory, the processor extracts a segment from the complete chromatographic elution curve of the sample, starting from the absorbance value at the end of the current sample measurement, up to the baseline, as a concentration-diffusion curve based on the absorbance value at the end of the current sample measurement, and uses this segment to correct the chromatographic elution curve of the next sample introduced at the end of the current measurement, and calculates the content of the analyte based on the chromatographic peak area corresponding to the analyte in the corrected chromatographic elution curve of the sample; wherein, Based on the concentration diffusion curve, the absorbance value of the chromatographic elution curve of the next sample introduced at the current measurement end time point is subtracted from the absorbance of the sample residue to correct the chromatographic elution curve of the next sample.

17. The sample analyzer as described in claim 16, characterized in that, The complete chromatographic elution curve is obtained by simulating the detection conditions of the sample analyzer and the preset sample diffusion law.

18. The sample analyzer as described in claim 16, characterized in that, The complete chromatographic elution curve is obtained by recording the chromatographic elution curve of the sample after all components have been detected and the sample has returned to the baseline using a sample analyzer in a non-continuous sample addition manner.

19. The sample analyzer as described in claim 16, characterized in that, The processor corrects the chromatographic elution curve of the next sample by subtracting the concentration diffusion curve from the chromatographic elution curve of the next sample measured at the current measurement end time point from the starting time point.

20. The sample analyzer as described in claim 16, characterized in that, The processor corrects the chromatographic elution curve of the current sample by using the concentration diffusion curve to partially complete the chromatographic elution curve of the current sample from the measurement end point to the return baseline.

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