Ghost peak trapping column, liquid chromatography system, analysis method and performance management method

Abstract

The invention provides a ghost peak trapping column for high performance liquid chromatography, a liquid chromatography system, a liquid chromatography analysis method and a performance management method. The ghost peak trapping column has the advantages that the stability of the ghost peak trapping column in a high-pressure environment is improved, the solvent mixing uniformity is optimized to improve the trapping efficiency, and the corrosion resistance is enhanced to prolong the service life.

Description

Technical Field

[0001] This application relates to the field of high performance liquid chromatography (HPLC) analysis technology, and more specifically, to an HPLC ghost peak collection column, an HPLC system, an HPLC analysis method, and a performance management method. Background Technology

[0002] In liquid chromatography (LC) analysis, especially during gradient elution operations or long-term continuous instrument operation, non-target peaks, or "ghost peaks," frequently appear in the chromatogram. These ghost peaks mainly originate from residual organic or inorganic impurities in the mobile phase, leaching of contaminants from the inner walls of the chromatographic system tubing, and exogenous substances introduced during sample pretreatment. The presence of ghost peaks hinders the analytical process, potentially causing signal distortion due to overlap with target analyte peaks, baseline drift, increased noise levels, and peak shape distortion, thereby affecting the accuracy of quantitative results and the reproducibility of the method. Such problems often force researchers to repeatedly adjust analytical conditions, re-validate method parameters, or even completely rebuild the analytical workflow, extending the method development cycle and increasing experimental costs.

[0003] Existing ghost peak collection column technology faces multiple technical bottlenecks in practical applications. First, its pressure tolerance is limited. The structural strength of conventional ghost peak collection columns is insufficient to withstand the high-pressure environment of ultra-high performance liquid chromatography (UHPLC) systems. Column deformation, seal failure, or joint leakage easily occur when operating pressure increases. This not only limits its applicability to UHPLC equipment but also prevents smooth transfer of analytical methods between conventional and UHPLC platforms, increasing the complexity and uncertainty of method development. Second, its internal fluid dynamics design is inadequate. The column head and tube structure of ghost peak collection columns fail to effectively guide uniform solvent distribution, resulting in flow rate differences, localized eddies, or dead volume regions when the mobile phase passes through the adsorbent material layer. This uneven solvent mixing not only weakens impurity collection efficiency but also easily induces bubble formation, generating additional bubble peak interference and further compromising baseline stability. Furthermore, their service life is generally short. Due to the insufficient corrosion resistance of the column material to strong acid and strong base mobile phases, and the susceptibility of the adsorbent material to chemical erosion or physical degradation during repeated use, existing ghost peak collection columns often exhibit problems such as adsorption capacity decay, abnormal column pressure increases, or background noise accumulation after a short period of operation, requiring frequent replacement. This increases daily operating costs and affects the continuity and reliability of analytical work. These shortcomings, combined, restrict the accuracy, efficiency, and economy of chromatographic analysis. Summary of the Invention

[0004] The purpose of this application is to provide a ghost peak collection column for high performance liquid chromatography, a liquid chromatography system, a liquid chromatography analysis method, and a performance management method, which has the advantages of improving the stability of the ghost peak collection column under high pressure environment, optimizing solvent mixing uniformity to improve collection efficiency, and enhancing corrosion resistance to extend service life.

[0005] To solve the above-mentioned technical problems, the present invention provides a ghost peak collection column, comprising: The column tube is made of corrosion-resistant material, and has an inner diameter of 4.6 mm and a length of 50-100 mm. An adsorbent material is packed inside the column tube, and the adsorbent material includes at least a covalent organic framework (COF) adsorbent with a particle size of 20-50 micrometers and activated carbon; A column head is disposed at both ends of the column tube, and the internal structure of the column head is configured to promote uniform mixing of the solvent; Metal pipe fitting, connected to the column head; The overall structural design of the ghost bee trap column is designed to withstand pressures of no less than 100 MPa.

[0006] As a further improvement of the present invention, the adsorbent material is a mixture of covalent organic framework (COF) adsorbent and activated carbon, wherein the mass percentage of covalent organic framework (COF) adsorbent in the mixture is 20% to 100%.

[0007] As a further improvement of the present invention, the loading amount of the adsorbent material is 0.415g to 0.833g.

[0008] As a further improvement of the present invention, the column tube is made of 316L stainless steel, and the metal pipeline joint is a special high-pressure sealing joint.

[0009] To achieve the above-mentioned objectives, the present invention also provides a liquid chromatography system, comprising: Mobile phase transport unit; As mentioned above, the ghost peak collection column is connected in series downstream of the mobile phase transport unit; An analytical chromatographic column, wherein the analytical chromatographic column is connected in series downstream of the ghost peak collecting column; A detector is used to detect the signals of components separated by the analytical chromatographic column.

[0010] To achieve the above-mentioned objectives, the present invention also provides a liquid chromatography analysis method using the aforementioned ghost peak trapping column, comprising the following steps: S1: Install the ghost peak trapping column between the gradient mixer and the autosampler of the liquid chromatography system; S2: Rinse the ghost peak collection column with an 80% (v / v) methanol aqueous solution to activate the adsorption material; S3: Compensate the time parameters of the gradient elution procedure based on the volume of the ghost peak capture column; S4: Run the gradient elution program in a liquid chromatography system containing the ghost peak trapping column to collect and acquire chromatograms.

[0011] As a further improvement of the present invention, the liquid chromatography system further includes an analytical column connected in series upstream of the ghost peak collection column and a detector for detecting the component signal separated by the analytical column; when the detector is a mass spectrometry detector, the method further includes step S0: before formally analyzing the sample, running a blank gradient program to evaluate the mass spectrometry baseline noise level after connecting the ghost peak collection column.

[0012] To achieve the above-mentioned objectives, the present invention also provides a performance management method for managing the performance of the aforementioned ghost peak collection column. The ghost peak collection column is installed in a liquid chromatography system, which includes a mobile phase delivery unit connected in series upstream of the ghost peak collection column, a downstream analytical column connected in series with the ghost peak collection column, and a detector for detecting the signal of the components separated by the analytical column. The performance management method for the ghost peak collection column includes the following steps: A1: Collect the operating data of the ghost peak trapping column in the liquid chromatography system, the operating data including the cumulative mobile phase throughput volume and detector baseline signal; A2: Based on the aforementioned operational data, calculate the current saturation of the ghost peak trapping column using a predefined adsorption capacity consumption model; A3: When the current saturation reaches the preset first warning threshold, output a prompt message suggesting maintenance of the ghost peak capture column.

[0013] As a further improvement of the present invention, the construction or updating of the adsorption capacity consumption model is also based on the baseline noise features or trace ghost peak features extracted after analyzing the baseline signal of the detector.

[0014] By adopting the above technical solution, the present invention has the following beneficial effects: High pressure compatibility and seamless method transfer: The ghost bee trap column has an overall structure that can withstand pressures of no less than 100 MPa. Combined with 316L stainless steel column tubing and a dedicated high-pressure sealing connector, it ensures stable operation under the high pressure environment of UHPLC, enabling seamless transfer of analytical methods between HPLC and UHPLC systems and improving method versatility.

[0015] Highly efficient ghost peak elimination: COFs adsorbents (or mixtures thereof with activated carbon) are used as adsorption materials. The high specific surface area, regular pore structure and abundant surface functional groups of COFs adsorbents, combined with an optimized particle size of 20-50 micrometers, can efficiently and specifically adsorb organic and aqueous impurities in the mobile phase. The addition of activated carbon broadens the impurity adsorption range, achieving a synergistic effect of broad-spectrum and selective adsorption, and significantly improving the ghost peak elimination effect.

[0016] Baseline stability optimization: The column heads at both ends of the column have a special internal structure to promote uniform solvent mixing, avoid local eddies and dead volumes, and reduce the generation of bubble peaks; at the same time, the uniform solvent distribution ensures that the adsorbent material can play its full role, improves baseline stability under large gradient elution conditions, and enhances analytical reproducibility.

[0017] Long service life and low operating costs: 316L stainless steel column tubes have excellent resistance to acid and alkali corrosion, reducing the erosion of the column by the flow relative; the optimized packing amount of adsorbent material (0.415g-0.833g) ensures sufficient adsorption capacity, and the activation and regeneration process with 80% methanol aqueous solution by volume extends the service life of the ghost bee trapping column and reduces the replacement frequency and operating costs.

[0018] Standardized analysis process: Clearly defined installation location, activation steps, and gradient compensation operations form a standardized analysis process, reducing human error; baseline noise pre-assessment steps for mass spectrometer detectors adapt to high-sensitivity detection requirements and expand application scenarios.

[0019] Intelligent performance management: Based on the quantitative monitoring mechanism of operating data and adsorption capacity consumption model, it realizes accurate calculation and early warning of ghost peak collection column saturation, avoids problems of premature or untimely maintenance, and continuously ensures analysis accuracy and system operating efficiency. Attached Figure Description

[0020] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0021] Figure 1 This is a schematic diagram of the ghost bee trapping column of the present invention; Figure 2 For use Figure 1 A schematic diagram of the liquid chromatography system for capturing the ghost peak; Figure 3 For use Figure 1 Flowchart of the liquid chromatography method for collecting *Ghost Peak* using a column; Figure 4 This is a gradient elution chart of the HPLC system in Example 1; Figure 5 This is a gradient elution chart of the UHPLC system in Example 2; Figure 6 This is a gradient elution chart of the HPLC system in Example 3; Figure 7 This is a gradient elution chart of the HPLC system in Example 4; Figure 8 The names and structural diagrams of the COFs selected in the packing material of the ghost bee trapping column of this invention are shown. Detailed Implementation

[0022] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0024] Those skilled in the art should understand that the following specific embodiments or implementation methods are a series of optimized configurations listed to further explain the specific content of the invention. These configuration methods can be combined or used in conjunction with each other, unless the invention explicitly states that some or a specific embodiment or implementation method cannot be associated with or used in conjunction with other embodiments or implementation methods. Furthermore, the following specific embodiments or implementation methods are merely optimized configurations and are not intended to limit the scope of protection of the invention.

[0025] In traditional liquid chromatography (LC) analysis, especially during gradient elution or prolonged instrument use, the generation of non-target peaks (ghost peaks) primarily originates from impurities in the mobile phase, tubing contamination, or contaminants introduced during sample pretreatment. Existing ghost peak trapping columns lack sufficient pressure tolerance, making stable operation in ultra-high performance liquid chromatography (UHPLC) systems difficult, hindering seamless transfer of analytical methods between conventional HPLC and UHPLC equipment. Uneven solvent distribution further impacts separation efficiency and baseline stability, even generating bubble peaks. Furthermore, the materials or design of ghost peak trapping columns are susceptible to corrosion by strong acid and alkali solvents, or the adsorbent material deteriorates rapidly, shortening their lifespan and reducing the accuracy and efficiency of chromatographic analysis.

[0026] For example, in gradient elution scenarios using a mobile phase containing buffer salts, impurities in the buffer salts manifest as ghost peaks on the chromatogram, interfering with the detection of the target analyte. Ghost peak trapping columns are prone to structural failure under high pressure conditions, and uneven solvent mixing leads to baseline fluctuations and bubble peak formation. Strong acid and base solvents accelerate the degradation process of the column material. Consequently, the reproducibility of chromatographic analysis is compromised, the separation between target and non-target peaks decreases, baseline noise increases, and the reliability of the analytical procedure is affected.

[0027] If the above problems are not addressed, the accuracy of chromatographic analysis will be continuously affected. The development of analytical methods may require repeated validation, increasing experimental complexity and potentially leading to erroneous analytical conclusions. Furthermore, frequent replacement of ghost peak collection columns will increase system maintenance frequency, limit the applicability of analytical methods across different equipment platforms, and further reduce overall analytical efficiency.

[0028] In this regard, such as Figure 1 As shown, the high-performance liquid chromatography ghost peak trapping column provided by the present invention mainly includes a column tube 100, an adsorption material 200, a column head 300, and a metal pipeline connector 400.

[0029] The column tube 100 is made of a corrosion-resistant material, with an inner diameter of 4.6 mm, a length of 50-100 mm, and a volume of approximately 0.83-1.66 mL; preferably, the column tube 100 is made of 316L stainless steel. Of course, in addition to 316L stainless steel, other highly corrosion-resistant alloys can also be used, such as Hastelloy or titanium alloys, which provide excellent performance even in extreme corrosive environments; or ultra-pure stainless steel with a specially passivated inner wall can be used to further reduce metal ion leaching and improve corrosion resistance.

[0030] The adsorbent material 200 is packed inside the column tube 100. In this embodiment, the particle size range of the adsorbent material 200 is 20-50 micrometers. This particle size range is selected to ensure that the adsorbent bed has moderate permeability, which can both ensure the smooth passage of the mobile phase and provide sufficient surface area for impurity adsorption.

[0031] Furthermore, the adsorbent 200 is a mixture of covalent organic framework (COF) adsorbent and activated carbon, wherein the mass percentage of the COF adsorbent ranges from 20% to 100%. Furthermore, the covalent organic framework (COF) adsorbent can be a vinyl COF with a uniform porous structure, preferably one of COF3, COF-701, V-COF-1, NKCOF-12, or NKCOF-62 (see [link to names and corresponding structures]). Figure 8 The preferred pore size is 0.6-1.8 nm, and the preferred specific surface area is 400-1700 m². 2 / g. High-specific-surface-area coconut shell activated carbon can be selected. The loading amount of adsorbent 200 is controlled between 0.415g and 0.833g, preferably 0.650g. This mixed adsorbent 200 combines the high selective adsorption of COFs materials with the broad-spectrum adsorption advantages of activated carbon.

[0032] The column head 300 is located at both ends of the column tube 100. Its internal structure (such as using a multi-channel distributor or a conical flow guide design) is specially designed to promote the uniform distribution and mixing of the mobile phase before entering the adsorbent material bed, thereby reducing flow path effects and bubble generation.

[0033] The metal pipeline connector 400 is connected to the column head 300. It is preferably a special high-pressure sealing connector with a conical seal or zero dead volume design to ensure reliable connection and no leakage under high pressure.

[0034] The metal pipeline connector is a specialized high-pressure sealing connector. This connector is a connection component designed to withstand high-pressure fluid systems. Its internal structure and materials are optimized to ensure a reliable, leak-free connection under high pressure. Its function in high-performance liquid chromatography (HPLC) systems is to ensure the sealing of pipeline connections, prevent mobile phase leakage, maintain stable system pressure, and prevent the entry of external contaminants or the dissolution of internal components, thereby ensuring analytical accuracy and baseline stability. This connector can employ various structural designs such as conical seals, spherical seals, or surface seals, and is often used in conjunction with high-strength threads and pressure-resistant gaskets (such as PEEK or stainless steel gaskets) to enhance the sealing effect; alternatively, it can adopt an integrated compression fitting design, achieving self-sealing under high pressure through precise machining and material selection.

[0035] Through the above structural design, and by selecting high-strength materials and reliable connection technology, the collection column of the present invention can withstand a pressure of not less than 100 MPa and is fully compatible with UHPLC systems.

[0036] When using the ghost peak collection column of this application, it is first installed between the gradient mixer and the autosampler of the liquid chromatography system. After the mobile phase is pumped out and mixed in the gradient mixer, it first enters the ghost peak collection column. Here, impurities in the mobile phase, such as organic contaminants in the buffer salt or non-volatile residues in the solvent, are efficiently adsorbed and retained by the adsorbent material 200 containing covalent organic frameworks (COFs) with a particle size of 20-50 micrometers, packed inside the column tube 100. The unique pore structure and abundant surface functional groups of the COFs adsorbent in the adsorbent material 200 enable it to exhibit excellent collection capabilities for a variety of organic impurities.

[0037] Before entering the adsorbent bed, the mobile phase first passes through column heads 300 located at both ends of the column tube 100. The internal structure of the column heads 300 is configured to promote uniform solvent mixing, ensuring that the mobile phase enters the adsorbent bed 200 at a uniform flow rate and distribution, avoiding impurity penetration or baseline fluctuations caused by uneven flow paths. The mobile phase purified by the ghost peak trapping column has a significantly reduced impurity content and then enters the analytical column for separation.

[0038] The entire ghost bee trap column 100 is made of corrosion-resistant materials, such as high-strength polymers or special stainless steel. This allows the ghost bee trap column to withstand strong acid and alkali environments that may be present in the mobile phase, thus extending its service life. Meanwhile, the column tube 100's inner diameter of 4.6 mm and length of 50-100 mm are designed to ensure sufficient adsorption capacity while keeping the system back pressure within a reasonable range. The metal pipe fittings 400 connecting the ghost bee trap column to the system piping ensure good sealing under high-pressure operating conditions, preventing leakage and the introduction of external contaminants.

[0039] More importantly, the overall structure of this ghost peak collection column is designed to withstand pressures of no less than 100 MPa. This means that even under the high-pressure environment of an ultra-high performance liquid chromatography (UHPLC) system, the ghost peak collection column can operate stably without structural deformation or leakage, thus ensuring the stability and reproducibility of the analytical method.

[0040] like Figure 2 As shown, the present invention also provides a liquid chromatography system. The system includes a mobile phase delivery unit 500 (such as a high-pressure pump), a gradient mixer 600, a ghost peak trapping column as described in this invention (installed after the gradient mixer 600), an autosampler 700, an analytical column 800, and a detector 900, connected in series.

[0041] In this system, the mobile phase delivery unit 500 delivers phase A (10 mmol / L potassium dihydrogen phosphate) and phase B (acetonitrile) to the gradient mixer 600 in a preset ratio, and after being mixed evenly, they enter the ghost peak collection column.

[0042] When the mobile phase flows through the ghost peak trapping column, impurities (such as organic pollutants in the buffer salt and non-volatile residues in the solvent) are efficiently trapped by the adsorbent material 200. The purified mobile phase enters the autosampler 700 and is then injected into the analytical chromatographic column 800.

[0043] The analytical column 800 separates the target components in the sample, and the separated components enter the detector 900 in sequence. The detector 900 collects the component signals and converts them into chromatograms.

[0044] Because the ghost peak trapping column removes impurities from the mobile phase, the chromatogram output by detector 900 is free of ghost peak interference, with a stable baseline, making target peak identification and quantification more accurate.

[0045] Please see Figure 3 As shown, the present invention provides a liquid chromatography analysis method using the above-mentioned ghost peak trapping column, the specific steps of which are as follows: S1: Install the ghost peak trapping column between the gradient mixer 600 and the autosampler 700 of the liquid chromatography system; S2: Rinse the ghost peak collection column with an 80% (v / v) methanol aqueous solution to activate the adsorbent material 200; S3: Compensate the time parameters of the gradient elution procedure based on the volume of the ghost peak capture column; S4: Run the gradient elution program in a liquid chromatography system containing the ghost peak trapping column to collect and acquire chromatograms.

[0046] S1 specifically involves installing the ghost peak trapping column between the gradient mixer 600 and the autosampler 700 of the HPLC or UHPLC system.

[0047] S2 specifically involves rinsing the ghost peak trapping column with approximately 10-20 column volumes of an 80% methanol aqueous solution at a normal flow rate (e.g., 1 mL / min) to remove manufacturing residues, wet and activate the adsorbent material 200.

[0048] In S3, since the ghost peak trap column introduces an additional delay volume of approximately 0.83–1.66 mL, the start time or gradient slope of the gradient elution procedure needs to be adjusted accordingly during method development or transfer to compensate for the delay and ensure the reproducibility of retention time.

[0049] Specifically, when detector 900 is a mass spectrometer detector, a blank gradient program (without injection or with pure solvent) is run before the formal analysis to assess the mass spectrometry baseline noise level and confirm that the system (especially the trapping column) has not introduced new mass spectrometry background interference.

[0050] The following description will provide a detailed explanation of the liquid chromatography analysis method of this application through specific embodiments.

[0051] Example 1 In this embodiment, the ghost peak trapping column is used in the HPLC system, employing a SHIMSEN Superb Ⅱ C18 analytical column (5 μm); mobile phase A is a 10 mmol / L potassium dihydrogen phosphate aqueous solution, mobile phase B is acetonitrile, gradient elution is performed, and detector 900 is an ultraviolet detector with a detection wavelength of 210 nm.

[0052] Please see Figure 4 As shown, without the ghost peak trapping column installed, mobile phases A and B are mixed and then fed into the autosampler 700 at a flow rate of 1 mL / min through the ghost peak trapping column, resulting in 10 μL of mobile phase. The flow temperature of the mobile phase through the analytical column 800 is further controlled at 40 °C. When the detection chromatogram is obtained by the UV detector, the baseline of the chromatogram fluctuates significantly, and significant ghost peaks appear at specific retention times, interfering with the quantification of the target analyte.

[0053] Furthermore, a ghost peak catching column (NKCOF-12 covalent organic framework adsorbent, coconut shell activated carbon, COFs mass percentage of 40%, column size 4.6mm x 50mm) was installed in the HPLC system and filled with the ghost peak catching column of the present invention, which is simultaneously filled with a mixture of covalent organic framework adsorbent and activated carbon as adsorbent material 200. After installation, the ghost peak catching column of the present invention was activated and the HPLC system was run under the same conditions. It can be seen that the ghost peaks on the chromatogram were basically eliminated, the baseline became flat and stable, the target peak shape was improved, and the accuracy and precision of the quantitative results were significantly improved.

[0054] Example 2 In this embodiment, the ghost peak trapping column is used in a UHPLC system. A Shim-pack Scepter HD-C18 analytical column (1.9 μm) is used, with water as mobile phase A and acetonitrile as mobile phase B for gradient elution. Detector 900 is an ultraviolet detector with a detection wavelength of 220 nm.

[0055] Please see Figure 5As shown, without the ghost peak trapping column installed, mobile phases A and B are mixed and then delivered at a flow rate of 1 mL / min through the ghost peak trapping column to the autosampler 700, resulting in 10 μL of mobile phase. The flow temperature of the mobile phase through the analytical column 800 is further controlled at 30 °C. When the detection chromatogram is obtained by the UV detector, the baseline of the chromatogram fluctuates significantly, and significant ghost peaks appear at specific retention times, interfering with the quantification of the target analyte.

[0056] Furthermore, in the UHPLC system, a ghost peak catching column of the present invention, simultaneously filled with a mixture of covalent organic framework adsorbent and activated carbon as adsorbent material 200 (the covalent organic framework adsorbent used is NKCOF-12, the activated carbon used is coconut shell carbon, the COFs mass ratio is 40%, the column size is 4.6mm x 50mm, this ghost peak catching column can withstand high pressure, ensuring a stable baseline during gradient elution and improving the sensitivity of impurity detection), is installed, and the ghost peak catching column is located between the gradient mixer 600 and the autosampler 700; after installation, the ghost peak catching column of the present invention is activated, and the UHPLC system is run under exactly the same conditions. It can be seen that the ghost peaks on the chromatogram are basically eliminated, the baseline becomes flat and stable, the target peak shape is improved, and the accuracy and precision of the quantitative results are significantly improved.

[0057] Example 3 In this embodiment, the ghost peak trapping column is used in the HPLC system. A Wondasil C18-WR analytical column (5 μm) is used. The mobile phase A is 10 mmol / L ammonium acetate and the mobile phase B is acetonitrile. Gradient elution is performed. The detector 900 is a UV detector with a detection wavelength of 254 nm.

[0058] Please see Figure 6 As shown, a ghost peak catching column filled only with activated carbon (coconut shell activated carbon was used, and the column size was 4.6 mm × 50 mm) was installed in the HPLC system, and the ghost peak catching column was located between the gradient mixer 600 and the autosampler 700; after mixing mobile phase A and mobile phase B, 10 μL of mobile phase was delivered to the autosampler 700 through the ghost peak catching column at a flow rate of 1 mL / min; and the mobile phase flow temperature was further controlled to be 40 °C through the analytical column 800; when the detection chromatogram was obtained by the UV detector, the baseline of the chromatogram fluctuated significantly, and significant ghost peaks appeared at specific retention times, interfering with the quantification of the target analyte.

[0059] Furthermore, the ghost peak collection column 200 in the HPLC system, which is filled with a mixture of covalent organic framework adsorbent and activated carbon as the adsorbent material (NKCOF-12 is selected as the covalent organic framework adsorbent, coconut shell carbon is selected as the activated carbon, the mass ratio of COFs is 40%, and the column size is 4.6mm x 50mm), is used to replace the ghost peak collection column filled only with activated carbon in the HPLC system. After installation, the ghost peak collection column of the present invention is activated, and the HPLC system is run under exactly the same conditions. It can be seen that the ghost peaks generated by the change of mobile phase composition due to the gradient running program are basically eliminated, the baseline becomes flat and stable, the target peak shape is improved, and the accuracy and precision of the quantitative results are significantly improved.

[0060] Example 4 In this embodiment, the ghost peak trapping column is used in the HPLC system. A Wondasil C18-WR analytical column (5 μm) is used. The mobile phase A is 10 mmol / L ammonium acetate and the mobile phase B is acetonitrile. Gradient elution is performed. The detector 900 is a UV detector with a detection wavelength of 254 nm.

[0061] Please see Figure 7 As shown, a ghost peak collection column 200, simultaneously packed with a mixture of covalent organic framework adsorbent and activated carbon as adsorbent material, is installed in the HPLC system and located between gradient mixer 600 and autosampler 700. Mobile phases A and B are mixed and then delivered at a flow rate of 1 mL / min through the ghost peak collection column to autosampler 700, resulting in 10 μL of mobile phase. The mobile phase is further controlled to flow through the analytical column 800 at a column temperature of 40 °C. When the chromatogram is obtained using a UV detector, the baseline of the chromatogram fluctuates significantly, and prominent ghost peaks appear at specific retention times, interfering with the quantification of the target analyte.

[0062] To further verify the effect of using the ghost peak collection column 200, which is filled with a mixture of covalent organic framework adsorbent and activated carbon as adsorbent material 200 as described in this application, NKCOF-12 was selected as the covalent organic framework adsorbent and coconut shell activated carbon was selected as the activated carbon. Adsorbent material 200 with COF adsorbent mass ratios of 10%, 20%, 40%, and 100% was filled into ghost peak collection columns with column dimensions of 4.6 mm x 50 mm, and installed separately or side by side in the HPLC system. After installation, the ghost peak collection column of the present invention was activated, and the HPLC system was run under exactly the same conditions. Ghost peaks were observed on the chromatogram due to the change in mobile phase composition caused by the gradient running program. The elimination effect of ghost peak collection columns with different COF adsorbent addition amounts was compared.

[0063] As shown in the figure, when the mass ratio of COFs adsorbent is 10%, the removal effect of ghost peaks is not obvious. When the mass ratio of COFs adsorbent is 20%, 40%, and 100%, the ghost peaks are basically eliminated, the baseline becomes flat and stable, the target peak shape is improved, and the accuracy and precision of the quantitative results are significantly improved.

[0064] Furthermore, the present invention also provides a performance management method for managing the performance of ghost peak capture columns, for predictive maintenance.

[0065] Performance management methods for ghost peak trap columns include: A1: Collect the operating data of the ghost peak trapping column in the liquid chromatography system, the operating data including the cumulative mobile phase throughput volume and detector baseline signal; A2: Based on the aforementioned operational data, calculate the current saturation of the ghost peak trapping column using a predefined adsorption capacity consumption model; A3: When the current saturation reaches the preset first warning threshold, output a prompt message suggesting maintenance of the ghost peak capture column.

[0066] Specifically, A1 refers to the continuous acquisition of operational data of the trapping column during chromatographic analysis, including but not limited to the total volume of mobile phase that has passed through and the real-time baseline signal of the detector 900 (such as UV or MS).

[0067] In A2, the collected data is input into a predefined adsorption capacity consumption model. In addition to considering the cumulative volume, this model can also extract features such as the root mean square value of baseline noise, the energy of noise at specific frequencies, or the number / area of ​​trace ghost peaks (peaks with intensity below the conventional threshold) from the baseline signal through signal processing algorithms, and comprehensively calculate the current saturation of the trapping column.

[0068] Specifically, A3 involves the system monitoring and calculating the saturation level in real time. When the saturation reaches a preset first warning threshold (e.g., 70%), the system automatically alerts the user, suggesting that the collection column be flushed and regenerated or replaced, thus taking action before performance significantly degrades and avoiding analysis failure.

[0069] In summary, the ghost bee trapping column of this invention has an overall structure that can withstand pressures of not less than 100 MPa. Combined with a 316L stainless steel column tube and a dedicated high-pressure sealing connector, it ensures stable operation under the high-pressure environment of UHPLC, realizes seamless transfer of analytical methods between HPLC and UHPLC systems, and improves the versatility of the methods.

[0070] The ghost peak collection column uses COFs adsorbents (or a mixture thereof with activated carbon) as the adsorbent material. The high specific surface area, regular pore structure, and abundant surface functional groups of COFs adsorbents, combined with an optimized particle size of 20-50 micrometers, enable them to efficiently and specifically adsorb organic and aqueous impurities in the mobile phase. The addition of activated carbon broadens the impurity adsorption range, achieving a synergistic effect of broad-spectrum and selective adsorption, and significantly improving the ghost peak elimination effect.

[0071] Furthermore, the column heads at both ends of the ghost peak trapping column have a special internal structure to promote uniform solvent mixing, avoid local eddies and dead volumes, and reduce the generation of bubble peaks. At the same time, the uniform solvent distribution ensures that the adsorbent material can play its full role, improves baseline stability under large gradient elution conditions, and enhances analytical reproducibility.

[0072] The ghost bee trap column uses 316L stainless steel tubing, which has excellent resistance to acid and alkali corrosion, reducing the erosion of the column by the flow phase. The optimized packing amount of adsorbent material (0.415g-0.833g) ensures sufficient adsorption capacity. Combined with the activation and regeneration process of 80% methanol aqueous solution, the service life of the ghost bee trap column is extended, and the replacement frequency and operating costs are reduced.

[0073] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A ghost beetle trapping column, characterized in that, include: The column tube is made of corrosion-resistant material, and the inner diameter of the column tube is 2.1-4.6mm and the length is 50-100mm. An adsorbent material is packed inside the column tube, and the adsorbent material includes at least a covalent organic framework adsorbent with a particle size of 20-50 micrometers and activated carbon. A column head is disposed at both ends of the column tube, and the internal structure of the column head is configured to promote uniform mixing of the solvent; Metal pipe fitting, connected to the column head; The overall structural design of the ghost bee trap column is designed to withstand pressures of no less than 100 MPa.

2. The ghost bee trapping column according to claim 1, characterized in that, The adsorbent material is a mixture of the covalent organic framework adsorbent and activated carbon, wherein the mass percentage of the covalent organic framework adsorbent in the mixture is 20% to 100%.

3. The ghost bee trapping column according to claim 2, characterized in that, The amount of the adsorbent material is between 0.415g and 0.833g.

4. The ghost bee trapping column according to any one of claims 1 to 3, characterized in that, The column tube is made of 316L stainless steel, and the metal pipeline joint is a special high-pressure sealing joint.

5. A liquid chromatography system, characterized in that, include: Mobile phase transport unit; The ghost peak collection column as described in any one of claims 1 to 4, wherein the ghost peak collection column is connected in series downstream of the mobile phase transport unit; An analytical chromatographic column, wherein the analytical chromatographic column is connected in series downstream of the ghost peak collecting column; A detector is used to detect the signals of components separated by the analytical chromatographic column.

6. A liquid chromatography analysis method using a ghost peak trapping column as described in any one of claims 1 to 4, characterized in that, Includes the following steps: S1: Install the ghost peak trapping column between the gradient mixer and the autosampler of the liquid chromatography system; S2: Rinse the ghost peak collection column with an 80% (v / v) methanol aqueous solution to activate the adsorption material; S3: Compensate the time parameters of the gradient elution procedure based on the volume of the ghost peak capture column; S4: Run the gradient elution program in a liquid chromatography system containing the ghost peak trapping column to collect and acquire chromatograms.

7. The liquid chromatography analysis method according to claim 6, characterized in that, The liquid chromatography system further includes an analytical column connected in series upstream of the ghost peak collection column and a detector for detecting the signal of the components separated by the analytical column; when the detector is a mass spectrometer detector, the method further includes step S0: before formally analyzing the sample, running a blank gradient program to evaluate the mass spectrometry baseline noise level after connecting the ghost peak collection column.

8. A performance management method for managing the performance of the ghost peak capture column according to any one of claims 1 to 4, characterized in that, The ghost peak catching column is installed in a liquid chromatography system, which includes a mobile phase delivery unit connected in series upstream of the ghost peak catching column, a downstream analytical column connected in series with the ghost peak catching column, and a detector for detecting the signal of the components separated by the analytical column. The performance management method for the ghost peak catching column includes the following steps: A1: Collect the operating data of the ghost peak trapping column in the liquid chromatography system, the operating data including the cumulative mobile phase throughput volume and detector baseline signal; A2: Based on the aforementioned operational data, calculate the current saturation of the ghost peak trapping column using a predefined adsorption capacity consumption model; A3: When the current saturation reaches the preset first warning threshold, output a prompt message suggesting maintenance of the ghost peak capture column.

9. The performance management method according to claim 8, characterized in that, The construction or updating of the adsorption capacity consumption model is also based on the baseline noise features or trace ghost peak features extracted after analyzing the baseline signal of the detector.

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