Phosphatidylcholine purity detection method and system
By constructing a multidimensional correlation model based on column temperature, pressure, and flow rate, optimizing gradient elution conditions, and selecting the optimal chromatogram, the problem of low detection accuracy of phosphatidylcholine purity under traditional gradient elution processes was solved, achieving higher detection accuracy.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HEBEI MERSWAY BIO-TECH CO LTD
- Filing Date
- 2026-02-06
- Publication Date
- 2026-04-28
AI Technical Summary
Traditional fixed gradient elution processes result in low accuracy in phosphatidylcholine purity detection, failing to effectively separate and identify the various components of phosphatidylcholine, thus affecting the accuracy of the detection results.
By combining column temperature change data, column pressure change data, and mobile phase flow rate data, a multidimensional correlation model was constructed to determine the elution accuracy index of the target experimental group, optimize gradient elution conditions, and use the local maximum peak point and peak symmetry response in the chromatogram to screen out the optimal chromatogram and obtain the purity result of phosphatidylcholine.
The accuracy of phosphatidylcholine purity detection has been improved, the error has been reduced, and higher detection accuracy has been achieved, with the error reduced from ±3.4% to ±1.2%.
Smart Images
Figure CN121933671A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of purity detection technology, specifically to a method and system for detecting the purity of phosphatidylcholine. Background Technology
[0002] Phosphatidylcholine (PC) is one of the main components of phospholipids and is widely used in food, pharmaceuticals, and health products. Purity testing ensures that the PC content in raw materials meets standards, such as the PC content in soybean lecithin needing to reach a specific ratio; it also avoids interference from impurities, such as lysophosphatidylcholine and free fatty acids, which can affect product performance.
[0003] High-performance liquid chromatography (HPLC) is a commonly used method for determining the purity of phosphatidylcholine in phospholipid solutions. When the sample enters the chromatographic column with the mobile phase, different components have different retention times due to varying degrees of interaction with the stationary phase. Chromatograms are then constructed based on the retention times and peak heights of each component in the solution. The purity of phosphatidylcholine in the phospholipid solution is obtained by comparing the chromatogram with a standard solution of the component to be detected under the same conditions.
[0004] High-performance liquid chromatography (HPLC) employs gradient elution to prepare the mobile phase mixture ratio. If the mobile phase contains a weakly polar A phase and a strongly polar B phase, in traditional gradient elution, the proportion of A phase is linearly increased from 0.1 to 0.9, while the proportion of B phase changes inversely. The purity is then determined by analyzing the chromatogram obtained under this gradient. However, in real-world scenarios, the content and types of components such as phosphatidylcholine in the test solution are unknown. Using a pre-set, fixed elution gradient may reduce the elution compatibility of each component. If the proportion of the weakly polar phase is too low in the initial stage of the gradient, weakly retained components may not be fully separated and may be flushed out, thus reducing the accuracy of phosphatidylcholine purity detection. Summary of the Invention
[0005] To address the technical problem of low accuracy in phosphatidylcholine purity detection using a pre-set fixed elution gradient preparation process, the present invention aims to provide a method and system for phosphatidylcholine purity detection. The specific technical solution adopted is as follows: This invention provides a method for determining the purity of phosphatidylcholine, the method comprising: During the gradient elution of phospholipid sample solutions, the elution accuracy index of the target experimental group was determined using column temperature change data, column pressure change data, and mobile phase flow rate data. The local maximum peak point in the chromatogram of the target experimental group is used to determine the local analytical peak in the chromatogram, and the peak symmetry response is determined by the slope change data of the analytical peak and the chromatographic data. The inter-peak resolution was determined by using the positional relationship data between the analytical peak and its adjacent analytical peaks, and the chromatogram optimization of the target experimental group was determined by using peak symmetry response, inter-peak resolution and elution accuracy indicators. The chromatogram corresponding to the experimental group with the highest chromatogram optimization degree was used as the optimal chromatogram for purity analysis, and the purity results of phosphatidylcholine were obtained.
[0006] Furthermore, the determination of the elution accuracy index of the target experimental group using column temperature change data, column pressure change data, and mobile phase flow rate data includes: The column temperature stability factor of the target experimental group was determined by using the column temperature data and the average column temperature. The temperature and pressure excellence coefficients of the target experimental group were determined using column temperature stability factor and column pressure change data of the chromatographic column. Using the temperature and pressure coefficients and the flow rate data of the mobile phase, the elution accuracy index of the target experimental group was determined.
[0007] Furthermore, the determination of the temperature and pressure excellence coefficient of the target experimental group using column temperature stability factor and column pressure change data includes: Determine the curve of the change in the proportion of the strongly polar phase in the mobile phase during the elution process for the target experimental group; Determine the correlation coefficient between the column pressure change curve and the proportion change curve corresponding to the column pressure change data of the chromatographic column; The temperature and pressure excellence coefficients of the target experimental group were determined using the column temperature stability factor and correlation coefficient.
[0008] Furthermore, the step of determining the elution accuracy index of the target experimental group using column temperature change data, column pressure change data, and mobile phase flow rate data further includes: Using the local maximum peak point in the chromatogram of the target experimental group as the plate, we can obtain the number of all plates in a single group and the average number of plates in all experimental groups in the chromatogram of the target experimental group. The elution accuracy index of the target experimental group was corrected by using the number of all trays and the average number of trays in a single group, and the corrected elution accuracy index was obtained.
[0009] Furthermore, the method of determining the local analytical peaks in the chromatogram using the local maximum peak points in the chromatogram of the target experimental group includes: Determine the local maximum peak point and the nearest local minimum point on both sides of it in the chromatogram of the target experimental group; The chromatographic range between the local maximum peak and the nearest local minimum on both sides is taken as the local analytical peak of the chromatogram.
[0010] Furthermore, the determination of peak symmetry reflectance using analytical peak slope change data and chromatographic data includes: The peak is segmented by the peak point of the analytical peak to obtain the slope of the first segment and the slope of the second segment. The difference between the slope of the first segment and the slope of the second segment is used to determine the peak tailing performance of the analytical peak. The chromatographic values spread outward from the peak point of the analytical peak at the same speed to both sides, and the same moment when chromatographic values exist on both sides is taken as the bilateral value segment; The peak symmetry response of the analytical peak is determined by using the peak tailing performance, the chromatographic difference corresponding to the bilateral value segment, and the proportion of the bilateral value segment.
[0011] Furthermore, determining the interpeak separation degree using the positional relationship data between the analytical peak and its adjacent analytical peaks includes: Determine the difference in the x-coordinate between the peak values of the analytical peak and its adjacent analytical peaks; Determine the difference between the ordinate of the midpoint of the intersection of the analytical peak and the midpoint of the adjacent analytical peak and the average ordinate of the two endpoints of the midpoint; The interpeak separation between an analytical peak and its adjacent analytical peaks is determined by using the difference between the horizontal and vertical axes.
[0012] Furthermore, the determination of the chromatographic optimization of the target experimental group using peak symmetry reactivity, interpeak resolution, and elution accuracy indicators includes: Peak analysis degradation indices for analytical peaks are determined by using peak symmetry reflectance and inter-peak separation. The chromatogram optimization of the target experimental group was determined by using elution accuracy index and peak analysis degradation index.
[0013] Furthermore, the step of using the chromatogram corresponding to the experimental group with the highest chromatogram optimization degree as the optimal chromatogram for purity analysis to obtain the purity result of phosphatidylcholine includes: The phosphatidylcholine standard solution was eluted according to the gradient elution conditions of the experimental group corresponding to the optimal chromatogram, and the highest peak in the eluted chromatogram was used as the standard peak. Obtain the retention time of the standard peak, take the peak to which the retention time of the standard peak in the optimal chromatogram belongs as the detection medium peak, and obtain the actual retention time corresponding to the detection medium peak; Determine the peak height difference between the actual peak height at the actual retention time in the detection medium and the peak height at the retention time of the standard peak; By using the retention time of the standard peak, the difference between the actual retention time and the peak height, the target chromatographic peak height corresponding to phosphatidylcholine in the phospholipid sample solution is determined, and the purity result of phosphatidylcholine is obtained based on the target chromatographic peak height.
[0014] The present invention also provides a purity detection system for phosphatidylcholine, the system being used to implement the purity detection method for phosphatidylcholine as described in any of the preceding claims; the system comprising: The elution accuracy analysis module is used to determine the elution accuracy index of the target experimental group by using column temperature change data, column pressure change data and mobile phase flow rate data during gradient elution of phospholipid sample solutions. The chromatogram analysis module is used to determine the local analytical peaks in the chromatogram of the target experimental group by using the local maximum peak points in the chromatogram; to determine the peak symmetry response by using the slope change data of the analytical peaks and chromatographic data; to determine the inter-peak resolution by using the positional relationship data between the analytical peaks and their adjacent analytical peaks; and to determine the chromatogram optimization of the target experimental group by using the peak symmetry response, inter-peak resolution and elution accuracy indicators. The component purity output module is used to select the chromatogram corresponding to the experimental group with the highest chromatogram optimization degree as the optimal chromatogram for purity analysis, and obtain the purity result of phosphatidylcholine.
[0015] The present invention has the following beneficial effects: This invention constructs an elution accuracy model based on a three-dimensional correlation of column temperature, pressure, and flow rate. Multiple gradient elution experimental groups with varying mobile phase ratios are prepared for the phospholipid solution to be tested. For each gradient elution experimental group, the temperature and pressure performance coefficient is obtained based on the column's temperature and pressure performance. Then, combined with the flow rate uniformity performance, the elution accuracy index for each gradient elution process is obtained. Furthermore, considering that air bubbles trapped in the column reduce column efficiency and thus the number of effective plates in the chromatogram, the elution accuracy index is corrected based on the number of plates. Simultaneously, for a single peak in the chromatogram corresponding to the current gradient elution conditions, the tailing peak performance is obtained based on the difference in the smoothness of the transition between the peak's segments. Based on the tailing peak performance, the symmetry trend of the peak is analyzed to obtain peak pairs. The reactivity is then combined with the inter-peak resolution to obtain peak analysis degradation parameters. Furthermore, the chromatogram optimization degree under each elution condition is obtained by combining the peak analysis degradation parameter levels of each peak in the chromatogram with the elution accuracy index. The chromatogram optimization degree is a multi-dimensional evaluation of operating conditions and peak shape, rather than a single peak shape index. Based on the chromatogram optimization degree, the optimal chromatogram is selected, and the peak height corresponding to phosphatidylcholine in the sample solution is obtained by combining the performance of the standard phosphatidylcholine solution under the same gradient elution conditions. Compared with the traditional method of directly using the peak height of the corresponding chromatogram under preset gradient elution conditions, this invention can combine multi-gradient elution conditions and chromatogram performance for comparative analysis, obtaining more accurate peak heights and thus achieving more accurate purity detection results for phosphatidylcholine. Attached Figure Description
[0016] To more clearly illustrate the technical solutions and advantages in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0017] Figure 1 The flowchart illustrates the steps of a method for detecting the purity of phosphatidylcholine according to an embodiment of the present invention. Figure 2 This is a detailed flowchart of step S1 in a method for detecting the purity of phosphatidylcholine provided in an embodiment of the present invention. Figure 3 This is a detailed flowchart of the purity detection method for phosphatidylcholine provided in an embodiment of the present invention, after step S1. Figure 4 This is a detailed flowchart of step S2 in a method for detecting the purity of phosphatidylcholine provided in an embodiment of the present invention. Figure 5 This is a detailed flowchart of step S3 in a method for detecting the purity of phosphatidylcholine provided in an embodiment of the present invention. Figure 6 This is a detailed flowchart of step S4 in a method for detecting the purity of phosphatidylcholine provided in an embodiment of the present invention. Figure 7 This is a schematic diagram of the hardware operating environment of the phosphatidylcholine purity detection device involved in the embodiments of the present invention; Figure 8 This is a schematic diagram of the framework structure of the phosphatidylcholine purity detection system involved in the embodiments of the present invention. Detailed Implementation
[0018] To further illustrate the technical means and effects adopted by the present invention to achieve the intended purpose, the following, in conjunction with the accompanying drawings and preferred embodiments, details the specific implementation, structure, features, and effects of a method for detecting the purity of phosphatidylcholine according to the present invention. In the following description, different "one embodiment" or "another embodiment" do not necessarily refer to the same embodiment. Furthermore, specific features, structures, or characteristics in one or more embodiments can be combined in any suitable form.
[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.
[0020] Before proceeding with the various embodiments of the technical solution of this invention, the purpose of this invention and the specific scenarios it addresses will be briefly explained.
[0021] The purpose of this invention is to evaluate the elution conditions and chromatographic performance under different gradient elution conditions, and then extract the chromatographic peak height of phosphatidylcholine under the optimal elution gradient.
[0022] Specifically, this invention requires determining the chromatographic peak height of phosphatidylcholine in the sample solution to obtain its purity percentage. Therefore, the elution accuracy index is first obtained based on the temperature and pressure performance of the chromatographic column and the stability of the liquid phase flow during gradient elution. Then, based on the elution accuracy index, the symmetry and separation performance of each peak in the chromatogram are matched and analyzed to obtain the chromatogram optimization degree corresponding to the experimental group under different gradient elution conditions. The optimal chromatogram is then selected, and the final chromatographic peak height of phosphatidylcholine in the sample solution is obtained by comparing and analyzing the performance of the optimal chromatogram with that of the standard solution.
[0023] The specific scenario targeted by this invention is as follows: Phosphatidylcholine is a component in some foods, pharmaceuticals and health products. It is generally obtained by separating phospholipid solutions. High performance liquid chromatography (HPLC) is a commonly used method to detect the purity of phosphatidylcholine in phospholipid solutions. Its principle is that different components in the solution have different residence times when entering the stationary phase along with the mobile phase, thereby distinguishing and identifying phosphatidylcholine.
[0024] HPLC employs a gradient elution process, in which a weakly polar A phase and a strongly polar B phase exist within the mobile phase. Gradient elution involves adjusting the proportions of different liquid phases in the mobile phase during the detection process to elute the components in the solution. The purity of phosphatidylcholine is then determined by comparing the eluted chromatogram with that of a standard solution. However, the purity detection results obtained using traditional methods with preset fixed gradient elution conditions are not accurate (see the background technology of this invention for details). Therefore, this invention combines elution conditions under multiple gradient elution conditions with the analysis of matched chromatogram performance to obtain more accurate purity detection results for phosphatidylcholine.
[0025] The specific scheme of the purity detection method for phosphatidylcholine provided by the present invention will be described in detail below with reference to the accompanying drawings.
[0026] Example 1: For the method for determining the purity of phosphatidylcholine provided by this invention, please refer to [link to relevant documentation]. Figure 1 The diagram illustrates a flowchart of a method for detecting the purity of phosphatidylcholine according to an embodiment of the present invention.
[0027] The method for determining the purity of phosphatidylcholine includes: Step S1: During the gradient elution of the phospholipid sample solution, the elution accuracy index of the target experimental group is determined using the column temperature change data, column pressure change data, and mobile phase flow rate data of the chromatographic column. In this embodiment, firstly, different gradient elution conditions are set for the phospholipid sample solution (hereinafter referred to as "sample solution") to be tested for phosphatidylcholine purity, specifically: a) The gradient elution process uses normal phase chromatography, where the stationary phase is a polar silica column, and the mobile phase A (weakly polar phase) is hexane-isopropanol (3:4), which is weakly polar; the mobile phase B (strongly polar phase) is hexane-isopropanol-water (3:4:0.75), which is strongly polar. It should be noted that the terms "weakly polar," "weakly polar phase," "strongly polar," and "strongly polar phase" are specific and well-defined concepts in this field.
[0028] b) Divide the sample solution into 6 pre-set portions and set up 6 experimental groups for the sample solution. In each experimental group, the accuracy level of the proportion of phase A to phase B during the elution process increases successively, and the total elution time is set to 40 min. c) For the first experimental group, during the elution process, the total elution time was divided into 5 segments. In the first segment, phase A accounted for 0.1 and phase B accounted for 0.9. In the last segment, phase A accounted for 0.9 and phase B accounted for 0.1. During this period, phase A increased uniformly and phase B decreased uniformly between each segment, and the proportions of phase A and phase B remained unchanged within the same segment. d) Then, for the second experimental group, the total elution time was divided into 6 segments. The changes in the proportion of phase A and phase B at the beginning, end and during the elution process were the same as those in the first experimental group, until the last experimental group (and so on, with the total elution time in the last experimental group being divided into 11 segments). The specific gradient elution conditions mentioned above can be adjusted according to actual needs; they are provided here as examples only.
[0029] Subsequently, the sample solution was eluted and separated under different gradient conditions according to the gradient elution conditions set above. Then, the column temperature and pressure data during the elution process of each experimental group were read through the temperature and pressure sensing module; the mobile phase flow rate information during the elution process of each experimental group was read through the phase velocity monitoring module; the chromatograms after elution of each experimental group were read through an ultraviolet (UV) spectrometer, with the detection wavelength selected as 205-207nm (the sensitive band of phosphatidylcholine); the data read above were cleaned and preprocessed; finally, the preprocessed data were used for subsequent analysis and use. The above operations completed the preliminary preparation and data acquisition work.
[0030] Considering that the column temperature changes due to the laboratory ambient temperature during gradient elution of phospholipid sample solutions, and that changes in column temperature alter molecular diffusion and mass transfer efficiency, resulting in reduced chromatographic accuracy, and that ideally, column pressure should be positively correlated with changes in flowability, the presence of bubbles and particulate matter in the mobile phase can cause the actual pressure to deviate from the expected change pattern, and that bubble phenomena reduce the uniformity of liquid phase flow, further increasing the probability of peak distortion in the chromatogram, this embodiment combines gradient elution conditions to obtain the elution accuracy index of each experimental group during the elution process.
[0031] Specifically, please refer to Figure 2 Step S1, which uses column temperature change data, column pressure change data, and mobile phase flow rate data to determine the elution accuracy index of the target experimental group, includes: Step S11: Determine the column temperature stability factor of the target experimental group using the column temperature data and the average column temperature of the chromatographic column. In this embodiment, during the gradient elution of the phospholipid sample solution, the chromatographic column serves as the carrier of the stationary phase. The column temperature may vary due to the influence of ambient temperature. When the column temperature is unstable, the fluctuating column temperature will alter the diffusion and mass transfer efficiency of liquid molecules when the liquid phase in the mobile phase enters the column and interacts with the mobile phase, thereby reducing the elution accuracy of the mobile phase.
[0032] Therefore, for any single experimental group, designated as the target experimental group, the column temperature change data during the elution of each component of the sample solution is obtained, and the column temperature at the i-th sampling time at the start of the experiment is denoted as . The experiment obtained a total of Data collected at each sampling time point; mean column temperature during elution was [value missing]. Calculate the column temperature stability factor for the current analytical gradient elution experimental group. : In the formula, This represents an exponential function with the natural constant as its base, and the same applies below. The worse the temperature uniformity of the chromatogram at different times during elution, the more unstable the column temperature, indicating a greater likelihood of peak distortion in the chromatogram after elution.
[0033] Step S12: Using the column temperature stability factor and column pressure change data, determine the temperature and pressure excellence coefficient of the target experimental group. More specifically, step S12 includes: Determine the curve of the change in the proportion of the strongly polar phase in the mobile phase during the elution process for the target experimental group; Determine the correlation coefficient between the column pressure change curve and the proportion change curve corresponding to the column pressure change data of the chromatographic column; The temperature and pressure excellence coefficients of the target experimental group were determined using the column temperature stability factor and correlation coefficient.
[0034] In this embodiment, during gradient elution, the proportion of the highly polar B-phase mixture in the mobile phase gradually increases, leading to an increase in the overall viscosity of the mobile phase and consequently an increase in column pressure. Normally, the column pressure should show a positive correlation with the mobile phase preparation. However, when there are error factors such as air bubbles in the chromatographic column, the correlation between column pressure and changes in the mobile phase ratio will be disrupted, thereby increasing the analytical error of the elution chromatogram.
[0035] Therefore, for a single target experimental group, the time-series column pressure change data during the elution process are obtained to construct a column pressure change curve, and simultaneously, a time-series curve of the proportion of phase B is constructed. The Pearson correlation coefficient between the two curves is then calculated. Calculate the temperature and pressure excellence coefficient of the current analytical gradient elution experimental group. : In the formula, norm represents the normalization of the maximum and minimum values. The same applies to the following embodiments, and the corresponding value range is [0,1]. If the Pearson correlation coefficient between the column pressure and the change in mobile phase preparation during the elution process of the current analytical experimental group is larger, it indicates that the change in column pressure conforms more closely to the correlation law. At the same time, if the column temperature is more stable, it further indicates that the elution accuracy is better during the purity detection process.
[0036] Step S13: Using the temperature and pressure excellence coefficient and the flow rate data of the mobile phase, determine the elution accuracy index of the target experimental group.
[0037] Furthermore, bubbles may remain in the mobile phase due to insufficient degassing, causing them to occupy part of the pipeline volume in the flow path, reducing the actual effective flow rate, thus affecting the stability of the mobile phase flow rate and further reducing the elution accuracy.
[0038] Therefore, for a single experimental group, the time-domain data of the mobile phase velocity during the elution process are obtained, and the standard deviation of the velocity data during the elution process is calculated. Then, the elution accuracy index of the current analytical gradient elution experimental group was calculated. : In the formula, the more stable the flow rate and the better the temperature and pressure conditions during the elution process in the current analytical experimental group, the higher the accuracy of the gradient elution. Furthermore, standard deviation is not considered. The extreme case of 0, limited ≠0.
[0039] The above implementation methods can be used to calculate and record the elution accuracy index of each gradient elution experimental group during the elution process.
[0040] In one embodiment, please refer to Figure 3 After step S1, the method further includes: Step S101: Take the local maximum peak point in the chromatogram of the target experimental group as the plate, and obtain the number of all plates in a single group and the average number of plates in all experimental groups in the chromatogram of the target experimental group. Step S102: Correct the elution accuracy index of the target experimental group by using the number of all trays and the average number of trays in a single group, and obtain the corrected elution accuracy index.
[0041] In this embodiment, considering that poor performance during gradient elution can lead to peak distortion in the chromatogram after elution, which mainly manifests as a decrease in peak symmetry and inter-peak separation, this embodiment obtains the optimization parameters of the chromatograms in each gradient elution experimental group by matching the elution accuracy index and the peak presentation effect in the chromatogram, and then selects the optimal chromatogram for the purity detection and analysis of phosphatidylcholine.
[0042] First, the bubbles flow into the column with the mobile phase, which disrupts the homogeneity of the stationary phase, increases the probability of eddy diffusion, and leads to a decrease in the number of plates in the chromatogram and a deterioration in separation efficiency.
[0043] Therefore, for a single gradient elution experimental group, the chromatogram after elution is obtained, and all local maximum peak points within the chromatogram are obtained using a peak point monitoring algorithm (existing algorithm). Each local maximum peak point is then used as a single plate to obtain the total number of plates in the chromatogram of that experimental group. Let be the total number of trays in a single group, and calculate the average number of trays for all experimental groups. The elution accuracy of the current analytical experimental group was assessed by the number of plates in the chromatogram after elution. Adjustments were made to obtain the corrected elution accuracy index. : In the formula, th() represents the hyperbolic tangent function. If the number of plates in the chromatogram after elution of the current experimental group is higher, it reflects that the elution accuracy of the experimental group is better. Therefore, the corresponding elution accuracy index of the experimental group can be appropriately increased.
[0044] Step S2: Determine the local analytical peaks in the chromatogram of the target experimental group using the local maximum peak points, and determine the peak symmetry response using the slope change data of the analytical peaks and the chromatographic data. Specifically, step S2, which involves determining the local analytical peaks in the chromatogram of the target experimental group using the local maximum peak points, includes: Determine the local maximum peak point and the nearest local minimum point on both sides of it in the chromatogram of the target experimental group; The chromatographic range between the local maximum peak and the nearest local minimum on both sides is taken as the local analytical peak of the chromatogram.
[0045] Taking the chromatogram of the current analytical experimental group as an example, the maximum local peak points in the chromatogram are extracted by the above embodiment. Then, the chromatographic range formed by the single maximum local peak point and the single local minimum value on its left and right sides is determined as a single peak in the chromatogram and recorded as the analytical peak, thereby extracting all analytical peaks in the chromatogram.
[0046] Specifically, please refer to Figure 4 Step S2, which uses the slope change data and chromatographic data of the analytical peak to determine its peak symmetry reflectance, includes: Step S21: The analytical peak is segmented by the peak point to obtain the slope of the first segment and the slope of the second segment. The difference between the slope of the first segment and the slope of the second segment is used to determine the peak tailing performance of the analytical peak. Step S22: Spread from the peak point of the analytical peak to both sides at the same speed, and take the same time when chromatographic values exist on both sides as the bilateral value segment; Step S23: Determine the peak symmetry response of the analytical peak by using the peak tailing performance, the chromatographic difference corresponding to the bilateral value segment, and the proportion of the bilateral value segment.
[0047] In this embodiment, due to the uneven flow rate of the mobile phase in the elution process, the diffusion path of some sample components within the column is prolonged, leading to increased peak tailing in the chromatogram after elution. Therefore, a single peak in the current experimental group (target experimental group) is randomly selected as the current analytical peak, and the peak is segmented using the maximum value point (peak point) as the segmentation point. The absolute value of the slope of the first segment of the current analytical peak is calculated. and the absolute value of the slope in the latter part .
[0048] For example, calculate the peak tailing performance of the current analytical peak in the current experimental group. : In the formula, if the absolute value of the slope of the latter part of the current analytical peak is closer to the absolute value of the slope of the former part, it indicates that the chromatographic change of the latter part of the current peak is more gradual than that of the former part, and it indicates that the tailing of the current peak is stronger.
[0049] Furthermore, since different components have different distribution rates in the chromatogram, if the mobile phase flow rate is uneven and the retention time of the component on the stationary phase is prolonged, it will be reflected as a strong tailing in the chromatogram. If the mobile phase flow rate is uneven and the elution rate of the component in the mobile phase is too fast, it will be reflected as a leading peak. Both tailing peaks and leading peaks will reduce the symmetry of the chromatographic peaks, thereby introducing peak analysis errors.
[0050] Therefore, for the current analytical peak in the current experimental group, the peak spreads outwards at the same rate from its peak value. If the chromatographic value of the peak exists on both sides at a certain moment, the corresponding moment is determined as a two-sided value segment; otherwise, it is a one-sided value segment. The proportion of the two-sided value segment of the current peak is calculated. (That is, the proportion of time intervals corresponding to the bilateral value segment), and at the same time, the chromatographic values on both sides of the peak within the bilateral value segment are subtracted, and the differences corresponding to all time intervals within the bilateral value segment are summed to obtain the bilateral deviation factor of the peak. For example, calculate the peak symmetry response of the current analytical peak. : In the formula, if the proportion of the bilateral value segment of the current analysis peak is larger, the numerical difference between the symmetrical points within the bilateral value segment is smaller, and the peak tailing is smaller, then the symmetry of the analysis peak is better.
[0051] Step S3: Determine the inter-peak resolution using the positional relationship data between the analytical peak and its adjacent analytical peaks; determine the chromatogram optimization of the target experimental group using peak symmetry response, inter-peak resolution, and elution accuracy indicators. Specifically, please refer to Figure 5 Step S3, which determines the interpeak separation degree using the positional relationship data between the analytical peak and its adjacent analytical peaks, includes: Step S31: Determine the difference in abscissa between the peak value of the analytical peak and the peak value of its adjacent analytical peaks; Step S32: Determine the difference between the ordinate of the midpoint of the analytical peak and the midpoint of the adjacent analytical peak and the average ordinate of the two endpoints of the midpoint. Step S33: Use the difference in the horizontal axis and the difference in the vertical axis to determine the interpeak separation between the analytical peak and its adjacent analytical peaks.
[0052] In this embodiment, fluctuations in column temperature and pressure during elution will reduce the accuracy of the chromatographic column in tracking liquid molecules, which will be reflected in the chromatogram as increased inter-peak redundancy and decreased resolution.
[0053] Therefore, taking the analysis of the current peak and its neighboring peaks to the left (or right) as an example, the difference in the horizontal coordinate between the peak values of the current peak and its neighboring peaks to the left is obtained. Simultaneously, the absolute value of the difference between the average ordinate of the midpoint of the two peaks and the two endpoints (i.e., the endpoints of the two peaks located on either side of the midpoint, such as the starting point of the left peak and the ending point of the currently analyzed peak) is obtained. For example, calculate the peak separation between the current analytical peak and its left neighboring peak. : In the formula, if the distance between the current analysis peak and the peak on the left is greater, and the numerical level of the intersection point between the two peaks is more consistent with the levels of the two endpoints, it reflects that the overlap between the peak and the neighboring peak on the left is worse, which further indicates that the separation between the peak and the neighboring peak on the left is better.
[0054] It should be noted that the average of the interpeak separation between the current analytical peak and its two adjacent peaks can be taken as the final interpeak separation of the current analytical peak. This final interpeak separation can then be used as the basis for determining the final interpeak separation of the analytical peak. Further analysis and calculations will be performed.
[0055] Specifically, step S3, which uses peak symmetry reactivity, interpeak separation, and elution accuracy to determine the chromatogram optimization of the target experimental group, includes: Peak analysis degradation indices for analytical peaks are determined by using peak symmetry reflectance and inter-peak separation. The chromatogram optimization of the target experimental group was determined by using elution accuracy index and peak analysis degradation index.
[0056] In this embodiment, if the peak symmetry of the current analytical peak is reflected... The worse (smaller the value), the lower the peak separation. The greater the overlap in the peak characteristics (the smaller the value), the greater the analytical error and the higher the degradation index of the current analytical peak. Therefore, we will use an example to calculate the peak analysis degradation index of the current analytical peak. : Subsequently, the mean peak degradation index of all analytical peaks in the current experimental group chromatogram was calculated. The elution accuracy index of this experimental group The peak appearance in the chromatograms was matched and analyzed to obtain the chromatogram optimization degree of the current experimental group. : In the formula, the higher the elution accuracy index of the current experimental group, the better the elution conditions of the experimental group in the gradient elution process, and the better the internal peak performance of the chromatogram obtained after elution (mean value of peak analysis degradation index). The smaller the value, the better the optimization of the chromatogram corresponding to the experimental group.
[0057] Step S4: The chromatogram corresponding to the experimental group with the highest chromatogram optimization degree is taken as the optimal chromatogram for purity analysis, and the purity result of phosphatidylcholine is obtained.
[0058] The chromatogram corresponding to the experimental group with the highest chromatogram optimization degree is extracted and used as the optimal chromatogram for the purity analysis of phosphatidylcholine.
[0059] Specifically, please refer to Figure 6 Step S4 includes: Step S41: Elute the phosphatidylcholine standard solution according to the gradient elution conditions of the experimental group corresponding to the optimal chromatogram, and obtain the highest peak in the eluted chromatogram and use it as the standard peak. Step S42: Obtain the retention time of the standard peak, take the peak to which the retention time of the standard peak in the optimal chromatogram belongs as the detection medium peak, and obtain the actual retention time corresponding to the detection medium peak. Step S43: Determine the peak height difference between the actual peak height at the actual retention time in the detection medium peak and the peak height at the retention time of the standard peak. Step S44: Using the retention time of the standard peak, the difference between the actual retention time and the peak height, determine the target chromatographic peak height corresponding to phosphatidylcholine in the phospholipid sample solution, and obtain the purity result of phosphatidylcholine based on the target chromatographic peak height.
[0060] In this embodiment, considering the influence of gradient elution precision, the deformation of the chromatographic peak of the sample solution may cause a slight shift in the retention time in the chromatogram, which in turn causes analytical errors in the comparison between the chromatographic peak of the phosphatidylcholine standard solution and the chromatographic peak of the sample solution. Therefore, this step adjusts the peak height of the phosphatidylcholine corresponding to the sample solution by comparing the performance of the standard and sample chromatograms.
[0061] First, a 1 mg / mL phosphatidylcholine standard solution was selected and eluted under the same gradient elution conditions as the optimal chromatogram described above. The highest peak in the eluted chromatogram was recorded as the standard peak, and the retention time of the standard peak was determined. (The retention time is the information on the horizontal axis of the chromatogram).
[0062] It should be noted that since the retention time of the highest peak remains basically unchanged after elution of solutions of different concentrations in the same medium under the same gradient elution conditions, it is sufficient to take any concentration of standard solution for comparative analysis.
[0063] Similarly, the peak with the retention time of the standard peak in the optimal chromatogram of the phospholipid sample solution is obtained as the detection medium peak, and the actual retention time corresponding to the detection medium peak is also obtained. and obtain the actual retention time. Actual peak height and the peak height corresponding to the retention time of the standard peak in the detection medium peak. : If the standard peak retention time is closer to the actual retention time ( The smaller the value, the higher the reliability of the actual peak height; conversely, the greater the value, the greater the possibility of retention time distortion and drift in the actual peak height, indicating that the actual peak height should be within the range specified in the original text. Based on this, more emphasis is placed on the peak height corresponding to the standard peak retention time. Adjustments were made to obtain the final target chromatographic peak height. : In the formula, Indicates the actual peak height in the detection medium. Peak height at standard peak retention time The difference in peak height.
[0064] The above process allows us to determine the chromatographic peak height corresponding to phosphatidylcholine in the phospholipid sample solution. .
[0065] To facilitate understanding of the above embodiments of the present invention, and the obtained chromatographic peak heights Regarding the impact on purity results, and for the subsequent complete procedure used in phosphatidylcholine purity testing, a brief extension is provided here: Specific implementation: First, prepare multiple sets of phosphatidylcholine standard solutions with different concentrations, ranging from 0.1 to 2.0 mg / mL. Elute each set of standard solutions under the same gradient elution conditions as the experimental group corresponding to the optimal chromatogram, obtain the chromatograms corresponding to each concentration of standard solution, and extract the peak height of the highest peak in each chromatogram.
[0066] A standard curve of phosphatidylcholine concentration was plotted with the concentration of each standard solution on the x-axis and the maximum peak height of the corresponding chromatogram of each standard solution on the y-axis.
[0067] Furthermore, the peak height of the chromatographic peak corresponding to the sample solution is located in the phosphatidylcholine concentration standard curve, and the abscissa of the peak height is extracted. The phosphatidylcholine concentration corresponding to this abscissa is the phosphatidylcholine concentration in the sample solution. After processing and analysis by the embodiments of the present invention, the purity detection error can be reduced from ±3.4% to ±1.2%.
[0068] This invention prepares multiple gradient elution experimental groups with varying mobile phase ratios for the phospholipid solution to be tested. For each gradient elution experimental group, the temperature and pressure performance coefficient is obtained based on the column's temperature and pressure performance. Then, combined with the flow rate uniformity performance, the elution accuracy index for each gradient elution process is obtained. Furthermore, considering that air bubbles trapped in the column reduce column efficiency and thus decrease the effective plate number in the chromatogram, the elution accuracy index is corrected based on the plate number level. Simultaneously, for each individual peak in the chromatogram corresponding to the current gradient elution conditions, the tailing peak performance is obtained based on the difference in the smoothness of the transition between the peak's preceding and following segments. Based on the tailing peak performance, the symmetry trend of the peak is analyzed. The method involves obtaining peak symmetry reflectance, then combining peak separation performance to obtain peak analysis degradation parameters, and then combining the peak analysis degradation parameter levels and elution accuracy indicators of each peak in the chromatogram to obtain the chromatogram optimization degree under each elution condition. The optimal chromatogram is then selected, and the chromatographic peak height corresponding to phosphatidylcholine in the sample solution is obtained by combining the performance of standard phosphatidylcholine solution under the same gradient elution conditions. Compared with the traditional method of directly using the chromatographic peak height of the corresponding chromatogram under preset gradient elution conditions, this invention can combine multi-gradient elution conditions and chromatographic performance for comparative analysis to obtain chromatographic peak heights with better accuracy, thereby obtaining more accurate purity detection results for phosphatidylcholine.
[0069] Example 2: This invention also proposes a device for detecting the purity of phosphatidylcholine. The device can be a high-performance liquid chromatograph, a computer, a server, or a combination of multiple devices for data analysis and computation.
[0070] like Figure 7 As shown, Figure 7 This is a schematic diagram of the hardware operating environment of the phosphatidylcholine purity detection device involved in the embodiments of the present invention.
[0071] like Figure 7 As shown, the phosphatidylcholine purity detection device may include: a processor 1001, such as a CPU; a network interface 1004; a user interface 1003; a memory 1005; and a communication bus 1002. The communication bus 1002 is used to establish communication between these components. The user interface 1003 may include a display or an input unit such as a control panel; optionally, the user interface 1003 may also include a standard wired interface or a wireless interface. The network interface 1004 may optionally include a standard wired interface or a wireless interface (such as a Wi-Fi interface). The memory 1005 may be a high-speed RAM or a stable, non-volatile memory, such as a disk drive. Optionally, the memory 1005 may also be a storage device independent of the aforementioned processor 1001. The memory 1005, as a computer storage medium, may include a phosphatidylcholine purity detection program.
[0072] Those skilled in the art will understand that Figure 7 The hardware structure shown does not constitute a limitation on the device and may include more or fewer components than shown, or combine certain components, or have different component arrangements.
[0073] Continue to refer to Figure 7 , Figure 7 The memory 1005, which is a computer-readable storage medium, may include an operating system, a user interface module, a network communication module, and a purity detection program for phosphatidylcholine.
[0074] exist Figure 7 In this embodiment, the network communication module is mainly used to connect to the server and can communicate with the server for data; while the processor 1001 can call the phosphatidylcholine purity detection program stored in the memory 1005 and execute the steps in the above embodiments.
[0075] Based on the hardware structure of the phosphatidylcholine purity detection device described above, various embodiments of the phosphatidylcholine purity detection method of the present invention are implemented.
[0076] In addition, the present invention also provides a purity detection system for phosphatidylcholine, please refer to... Figure 8 The purity detection system for phosphatidylcholine includes: The elution accuracy analysis module A10 is used to determine the elution accuracy index of the target experimental group by using column temperature change data, column pressure change data and mobile phase flow rate data during gradient elution of phospholipid sample solutions. The chromatogram analysis module A20 is used to determine the local analytical peaks in the chromatogram of the target experimental group by using the local maximum peak points in the chromatogram; to determine the peak symmetry response by using the slope change data of the analytical peaks and the chromatographic data; to determine the inter-peak resolution by using the positional relationship data between the analytical peaks and their adjacent analytical peaks; and to determine the chromatogram optimization of the target experimental group by using the peak symmetry response, inter-peak resolution and elution accuracy indicators. The component purity output module A30 is used to select the chromatogram corresponding to the experimental group with the highest chromatogram optimization degree as the optimal chromatogram for purity analysis, and obtain the purity result of phosphatidylcholine.
[0077] Furthermore, the elution accuracy analysis module A10 is also used for: The column temperature stability factor of the target experimental group was determined by using the column temperature data and the average column temperature. The temperature and pressure excellence coefficients of the target experimental group were determined using column temperature stability factor and column pressure change data of the chromatographic column. Using the temperature and pressure coefficients and the flow rate data of the mobile phase, the elution accuracy index of the target experimental group was determined.
[0078] Furthermore, the elution accuracy analysis module A10 is also used for: Determine the curve of the change in the proportion of the strongly polar phase in the mobile phase during the elution process for the target experimental group; Determine the correlation coefficient between the column pressure change curve and the proportion change curve corresponding to the column pressure change data of the chromatographic column; The temperature and pressure excellence coefficients of the target experimental group were determined using the column temperature stability factor and correlation coefficient.
[0079] Furthermore, the elution accuracy analysis module A10 is also used for: Using the local maximum peak point in the chromatogram of the target experimental group as the plate, we can obtain the number of all plates in a single group and the average number of plates in all experimental groups in the chromatogram of the target experimental group. The elution accuracy index of the target experimental group was corrected by using the number of all trays and the average number of trays in a single group, and the corrected elution accuracy index was obtained.
[0080] Furthermore, the chromatogram analysis module A20 is also used for: Determine the local maximum peak point and the nearest local minimum point on both sides of it in the chromatogram of the target experimental group; The chromatographic range between the local maximum peak and the nearest local minimum on both sides is taken as the local analytical peak of the chromatogram.
[0081] Furthermore, the chromatogram analysis module A20 is also used for: The peak is segmented by the peak point of the analytical peak to obtain the slope of the first segment and the slope of the second segment. The difference between the slope of the first segment and the slope of the second segment is used to determine the peak tailing performance of the analytical peak. The chromatographic values spread outward from the peak point of the analytical peak at the same speed to both sides, and the same moment when chromatographic values exist on both sides is taken as the bilateral value segment; The peak symmetry response of the analytical peak is determined by using the peak tailing performance, the chromatographic difference corresponding to the bilateral value segment, and the proportion of the bilateral value segment.
[0082] Furthermore, the chromatogram analysis module A20 is also used for: Determine the difference in the x-coordinate between the peak values of the analytical peak and its adjacent analytical peaks; Determine the difference between the ordinate of the midpoint of the intersection of the analytical peak and the midpoint of the adjacent analytical peak and the average ordinate of the two endpoints of the midpoint; The interpeak separation between an analytical peak and its adjacent analytical peaks is determined by using the difference between the horizontal and vertical axes.
[0083] Furthermore, the chromatogram analysis module A20 is also used for: Peak analysis degradation indices for analytical peaks are determined by using peak symmetry reflectance and inter-peak separation. The chromatogram optimization of the target experimental group was determined by using elution accuracy index and peak analysis degradation index.
[0084] Furthermore, the component purity output module A30 is also used for: The phosphatidylcholine standard solution was eluted according to the gradient elution conditions of the experimental group corresponding to the optimal chromatogram, and the highest peak in the eluted chromatogram was used as the standard peak. Obtain the retention time of the standard peak, take the peak to which the retention time of the standard peak in the optimal chromatogram belongs as the detection medium peak, and obtain the actual retention time corresponding to the detection medium peak; Determine the peak height difference between the actual peak height at the actual retention time in the detection medium and the peak height at the retention time of the standard peak; By using the retention time of the standard peak, the difference between the actual retention time and the peak height, the target chromatographic peak height corresponding to phosphatidylcholine in the phospholipid sample solution is determined, and the purity result of phosphatidylcholine is obtained based on the target chromatographic peak height.
[0085] The specific implementation of the phosphatidylcholine purity detection system of the present invention is basically the same as the embodiments of the above-described phosphatidylcholine purity detection method, and will not be repeated here.
[0086] Furthermore, the present invention also provides a computer-readable storage medium. The computer-readable storage medium of the present invention stores a purity detection program for phosphatidylcholine, wherein, when executed by a processor, the purity detection program for phosphatidylcholine implements the steps of the purity detection method for phosphatidylcholine as described above.
[0087] The method implemented when the purity detection procedure for phosphatidylcholine is executed can be referred to in various embodiments of the purity detection method for phosphatidylcholine of the present invention, and will not be repeated here.
[0088] It should be noted that the order of the above embodiments of the present invention is merely for descriptive purposes and does not represent the superiority or inferiority of the embodiments. The processes depicted in the accompanying drawings do not necessarily require a specific or sequential order to achieve the desired result. In some embodiments, multitasking and parallel processing are also possible or may be advantageous.
[0089] The various embodiments in this specification are described in a progressive manner. The same or similar parts between the various embodiments can be referred to each other. Each embodiment focuses on describing the differences from other embodiments.
[0090] Those skilled in the art will understand that embodiments of the present invention can be provided as methods, systems, or computer program products. Therefore, the present invention can take the form of a completely hardware embodiment, a completely software embodiment, or an embodiment combining software and hardware aspects. Furthermore, the present invention can take the form of a computer program product embodied on one or more computer-usable storage media (including, but not limited to, disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.
[0091] The above description is only a preferred embodiment of the present invention and does not limit the scope of protection of the present invention. All equivalent structural / method transformations made under the inventive concept of the present invention using the contents of the present invention specification and drawings, or direct / indirect applications in other related technical fields, are included within the scope of protection of the present invention.
Claims
1. A method for determining the purity of phosphatidylcholine, characterized in that, The method includes: During the gradient elution of phospholipid sample solutions, the elution accuracy index of the target experimental group was determined using column temperature change data, column pressure change data, and mobile phase flow rate data. The local maximum peak point in the chromatogram of the target experimental group is used to determine the local analytical peak in the chromatogram, and the peak symmetry response is determined by the slope change data of the analytical peak and the chromatographic data. The inter-peak resolution was determined by using the positional relationship data between the analytical peak and its adjacent analytical peaks, and the chromatogram optimization of the target experimental group was determined by using peak symmetry response, inter-peak resolution and elution accuracy indicators. The chromatogram corresponding to the experimental group with the highest chromatogram optimization degree was used as the optimal chromatogram for purity analysis, and the purity results of phosphatidylcholine were obtained.
2. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The determination of the elution accuracy index for the target experimental group using column temperature change data, column pressure change data, and mobile phase flow rate data includes: The column temperature stability factor of the target experimental group was determined by using the column temperature data and the average column temperature. The temperature and pressure excellence coefficients of the target experimental group were determined using column temperature stability factor and column pressure change data of the chromatographic column. Using the temperature and pressure coefficients and the flow rate data of the mobile phase, the elution accuracy index of the target experimental group was determined.
3. The method for determining the purity of phosphatidylcholine according to claim 2, characterized in that, The determination of the temperature and pressure excellence coefficient of the target experimental group using column temperature stability factor and column pressure change data includes: Determine the curve of the change in the proportion of the strongly polar phase in the mobile phase during the elution process for the target experimental group; Determine the correlation coefficient between the column pressure change curve and the proportion change curve corresponding to the column pressure change data of the chromatographic column; The temperature and pressure excellence coefficients of the target experimental group were determined using the column temperature stability factor and correlation coefficient.
4. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The process of determining the elution accuracy index of the target experimental group using column temperature change data, column pressure change data, and mobile phase flow rate data also includes: Using the local maximum peak point in the chromatogram of the target experimental group as the plate, we can obtain the number of all plates in a single group and the average number of plates in all experimental groups in the chromatogram of the target experimental group. The elution accuracy index of the target experimental group was corrected by using the number of all trays and the average number of trays in a single group, and the corrected elution accuracy index was obtained.
5. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The method of determining the local analytical peaks in the chromatogram using the local maximum peak points in the chromatogram of the target experimental group includes: Determine the local maximum peak point and the nearest local minimum point on both sides of it in the chromatogram of the target experimental group; The chromatographic range between the local maximum peak and the nearest local minimum on both sides is taken as the local analytical peak of the chromatogram.
6. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The determination of peak symmetry response using analytical peak slope change data and chromatographic data includes: The peak is segmented by the peak point of the analytical peak to obtain the slope of the first segment and the slope of the second segment. The difference between the slope of the first segment and the slope of the second segment is used to determine the peak tailing performance of the analytical peak. The chromatographic values spread outward from the peak point of the analytical peak at the same speed to both sides, and the same moment when chromatographic values exist on both sides is taken as the bilateral value segment; The peak symmetry response of the analytical peak is determined by using the peak tailing performance, the chromatographic difference corresponding to the bilateral value segment, and the proportion of the bilateral value segment.
7. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The determination of the interpeak separation degree using the positional relationship data between the analytical peak and its adjacent analytical peaks includes: Determine the difference in the x-coordinate between the peak values of the analytical peak and its adjacent analytical peaks; Determine the difference between the ordinate of the midpoint of the intersection of the analytical peak and the midpoint of the adjacent analytical peak and the average ordinate of the two endpoints of the midpoint; The interpeak separation between an analytical peak and its adjacent analytical peaks is determined by using the difference between the horizontal and vertical axes.
8. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The determination of the chromatogram optimization of the target experimental group using peak symmetry reactivity, interpeak resolution, and elution accuracy indicators includes: Peak analysis degradation indices for analytical peaks are determined by using peak symmetry reflectance and inter-peak separation. The chromatogram optimization of the target experimental group was determined by using elution accuracy index and peak analysis degradation index.
9. The method for determining the purity of phosphatidylcholine according to claim 1, characterized in that, The process of using the chromatogram corresponding to the experimental group with the highest chromatogram optimization degree as the optimal chromatogram for purity analysis to obtain the purity result of phosphatidylcholine includes: The phosphatidylcholine standard solution was eluted according to the gradient elution conditions of the experimental group corresponding to the optimal chromatogram, and the highest peak in the eluted chromatogram was used as the standard peak. Obtain the retention time of the standard peak, take the peak to which the retention time of the standard peak in the optimal chromatogram belongs as the detection medium peak, and obtain the actual retention time corresponding to the detection medium peak; Determine the peak height difference between the actual peak height at the actual retention time in the detection medium and the peak height at the retention time of the standard peak; By using the retention time of the standard peak, the difference between the actual retention time and the peak height, the target chromatographic peak height corresponding to phosphatidylcholine in the phospholipid sample solution is determined, and the purity result of phosphatidylcholine is obtained based on the target chromatographic peak height.
10. A purity detection system for phosphatidylcholine, characterized in that, The system is used to implement the purity detection method for phosphatidylcholine as described in any one of claims 1 to 9; the system comprises: The elution accuracy analysis module is used to determine the elution accuracy index of the target experimental group by using column temperature change data, column pressure change data and mobile phase flow rate data during gradient elution of phospholipid sample solutions. The chromatogram analysis module is used to determine the local analytical peaks in the chromatogram of the target experimental group by using the local maximum peak points in the chromatogram; to determine the peak symmetry response by using the slope change data of the analytical peaks and chromatographic data; to determine the inter-peak resolution by using the positional relationship data between the analytical peaks and their adjacent analytical peaks; and to determine the chromatogram optimization of the target experimental group by using the peak symmetry response, inter-peak resolution and elution accuracy indicators. The component purity output module is used to select the chromatogram corresponding to the experimental group with the highest chromatogram optimization degree as the optimal chromatogram for purity analysis, and obtain the purity result of phosphatidylcholine.