Monolithic Column Research Articles

Segmented two-dimensional liquid chromatography. Proof of concept study

The separation in liquid chromatography is defined either by the space domain where it proceeds until the least retained analyte reaches the outlet of the column or by the time when individual analytes elute out of the column. These two approaches lead to the four possible combinations of two-dimensional liquid chromatography with online space x time coupling being the least experimentally feasible. Here, we show the development of a novel two-dimensional liquid chromatography method combining separation defined by space and the conventional elution-based separation. First-dimension column consisted of four capillary segments coupled serially via two-position six-port valves allowing an online and comprehensive transfer of analytes from the first to the second dimension. After initial experiments using homemade monolithic capillary columns, we tested commercially available columns in both dimensions. We ended with the combination of packed capillary columns in the first dimension and monolithic capillary column in the second dimension. We used a reversed-phase retention mechanism in the first spatial dimension, while HILIC was in the second, time-based dimension. We also developed a theoretical model to describe the proposed two-dimensional separation that was further confirmed by utilizing both an isocratic and gradient elution in the second dimension. Finally, we applied our experimental setup to separate neurotransmitters contained in human urine.

Seismic performance test research on a new type of replaceable energy-consuming joint column

This study proposed a new type of fully-precast reinforced concrete column-column joint, a replaceable energy-consuming joint (RECJ), which was composed of tenons and replaceable energy-dissipating connecting plates. The joint's seismic behavior and failure mechanism were investigated using quasi-static tests on two RECJ-connecting precast columns, one cast-in situ column, and one repaired RECJ-connecting precast column. The results showed that the damage and destruction of RECJ-connecting specimens were mainly concentrated on the connecting steel plates, while the other parts were basically elastic, showing a compression-bending failure mode. The hysteresis curves of RECJ-connecting specimens were fully shuttle-shaped. Meanwhile, energy dissipation, ductility, and stiffness degradation performance of these specimens were significantly better than those of the monolithic concrete comparison column. At the same time, the specimen with a longer tenon exhibited better overall seismic performance than that with a shorter tenon, while having the same initial stiffness and bearing capacity as the cast-in situ column. The seismic performance of the repaired long-tenon RECJ-connecting specimen with connecting plates replaced was not significantly decreased compared with original specimen, which demonstrated that RECJ had fine repairability. The relevant parameters were optimized through finite element simulation (FEM). Finally, based on test results, the shear capacity equation of RECJ was deduced according to the force equilibrium of an isolated body in the joint area and the accuracy of the equation verified.

Continuous Phosphate Removal and Recovery Using a Calcium Silicate Hydrate Composite Monolithic Cryogel Column.

Toward the development of a practical and green approach for removing phosphate from water, a monolithic cryogel based on starch and calcium silicate hydrate (Cry-CSH) was employed as a phosphate adsorbent in a continuous flow system for the first time. The influence of flow rate, initial phosphate concentration, and adsorbent height on the adsorption efficiency was investigated. As the rate of flow and the initial concentration of phosphate increased, the total quantity of adsorbed phosphate dropped; however, the performance of the column was greatly enhanced by an increase in adsorbent height. The experimental data fit the Adams-Bohart model better than the Thomas and Yoon-Nelson models at the beginning of the adsorption process. To evaluate its applicability, the continuous flow system based on the monolithic Cry-CSH column was applied for the removal of phosphate from the discharge effluent of the Patong Municipality Wastewater Treatment Plant (Phuket, Thailand), achieving an excellent total adsorption of 94.61%.

A monolithic azacalix[4]pyridine column for high-resolution and high-efficiency pressurized capillary electrochromatographic separation

Herein, a highly stable (NAllyl)4-bridged alyzacalix[4]pyridine monolithic column (nAC4P@capillary) was fabricated by copolymerizing nAC4P with GMA and EDMA into a stationary phase through a simple olefin addition reaction. The physicochemical properties of the prepared materials were characterized by scanning electron microscopy (SEM), thermogravimetric analysis (TGA), energy dispersive X-ray (EDS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and elemental analysis (EA). A wide range of analytes with various properties was well separated and demonstrated the synergistic effect of the various interaction sites. The maximum column efficiencies (Nmax) for all the analytes reached 62,368 plates/m, and the relative standard deviations (RSDs) of the daily retention times of alkylbenzenes, measured over six months were all lower than 5.63%. Non-covalent interaction analysis reveals that nAC4P@capillary can effectively combine the advantages of electronic configurations, π-π interactions, CH-π interactions, O–H…N hydrogen bonds and lone pair-π interactions, providing excellent prospects for high-performance chromatographic separation.

Bimetallic Cluster Cu7Zn2 Functionalized Copolymer Monolithic Column for Pipette Tip Micro-Solid Phase Extraction of Eight Phthalate Esters in Edible Oil

Bimetallic Cluster Cu7Zn2 Functionalized Copolymer Monolithic Column for Pipette Tip Micro-Solid Phase Extraction of Eight Phthalate Esters in Edible Oil

Nano-liquid chromatography with monolithic stationary phase based on naphthyl monomer for proteomics analysis

Monolithic poly(2-vinylnaphthalene-co-divinylbenzene) columns were introduced, for the first time, and were evaluated as the separation media for nano-liquid chromatography (nano-LC). These columns were prepared by in-situ polymerization of 2-vinylnaphthalene (2-VNA) as the functional monomer and divinylbenzene (DVB) as the crosslinker in a fused silica capillary column of 50 µm i.d. Various porogenic solvents, including tetrahydrofuran (THF), dodecanol and toluene were used for morphology optimization. Final monolithic column (referred to as VNA column) was characterized by using scanning electron microscopy (SEM) and chromatographic analyses. Alkylbenzenes (ABs), and polyaromatic hydrocarbons (PAHs) were separated using the VNA column while the column offered excellent hydrophobic and π-π interactions under reversed-phase conditions. Theoretical plates number up to 41,200 plates/m in isocratic mode for ethylbenzene could be achieved. The potential of the final VNA column was demonstrated with a gradient elution in the separation of six intact proteins, including ribonuclease A (RNase A), cytochrome C (Cyt C), lysozyme (Lys), β-lactoglobulin (β-lac), myoglobin (My) and α-chymotrypsinogen (α-chym) in nano LC system. The column was then applied to the peptide analysis of trypsin digested cytochrome C, allowing a high peak capacity up to 1440 and the further proteomics analysis of COS-7 cell line was attempted applying the final monolithic column in nano-LC UV system.

Lectin Purification through Affinity Chromatography Exploiting Macroporous Monolithic Adsorbents

Growing medical, engineering, biochemical, and biological interest has led to a steady pace of research and development into polymeric monolithic structures with densely interconnected pores for purifying bio compounds. Cryogels, which are generated by freezing a reactive polymerization mixture, are highlighted due to their versatility and low relative cost as macroporous, polymeric, monolithic adsorbents. The conversion of cryogels into affinity adsorbents is one possible alternative to their optimal application. Some of the most often utilized supports for immobilizing particular ligands are monolithic columns manufactured with epoxy radicals on their surfaces. The purification of biomolecules with a high degree of specificity, such as lectins and glycoproteins with an affinity for glycosylated groups, has garnered interest in the use of fixed non-traditional beds functionalized with ligands of particular interest. The interaction is both robust enough to permit the adsorption of glycoproteins and reversible enough to permit the dissociation of molecules in response to changes in the solution’s pH. When compared to other protein A-based approaches, this one has been shown to be more advantageous than its counterparts in terms of specificity, ease of use, and cost-effectiveness. Information on polymeric, macroporous, monolithic adsorbents used in the affinity chromatographic purification of lectins has been published and explored.

Removal of amoxicillin via chromatographic monolithic columns: comparison between batch and continuous fixed bed.

This study presented a hydrophobic interaction-based poly(HEMA-MATrp) monolithic chromatographic column (MCC) to remove amoxicillin from aqueous solutions. In addition to their porous structure, monolithic-filled columns offer superior properties without loss of performance, which is one of the points that make them unique. The specific surface area of the monolithic column synthesized by the bulk polymerization of 2-hydroxyethyl methacrylate and N-Methacryloyl-L-tryptophan. Also, poly(HEMA-MATrp) MCC has been characterized via FTIR, SEM, and elemental analysis. According to BET analysis, the specific surface area of the poly(HEMA-MATrp) monolithic chromatographic column (MCC) is 14.2 mg/g. The adsorption and desorption of amoxicillin in an aqueous solution were investigated comparatively in both continuous fixed bed and batch adsorption. The highest adsorption value of amoxicillin was determined at pH 7 in the presence of PBS as 62.11 mg/g. The appropriate adsorption isotherm for the adsorption of amoxicillin was Langmuir, and the reaction kinetics was pseudo-second-order. No significant loss was observed for the adsorption capacity of poly(HEMA-MATrp) MCC after the 5 cycles of adsorption-desorption studies. Also, the loss for the adsorption capacity of the monolithic column is just %5.2 after 6-month storage, proving the reusability and storability of the monolithic column.

Application of Metal-Organic Frameworks to High-Performance Liquid Chromatography

We present a critical review of experimental studies of the efficiency of the separation of various groups of chemical compounds under the conditions of high-performance liquid chromatography using metal-organic framework polymers (metal-organic frameworks (MOFs)) or composite materials with MOFs as a stationary phase. MOFs demonstrated many advantages in high-performance liquid chromatography and a possibility of competing successfully with conventional phases, especially in the determination of low-molecular-weight compounds and optical isomers. Among the promising options for using this class of materials are the development of monolithic separation columns and hybrid adsorbents that eliminate the disadvantages of adsorbents based on pure MOFs.

Automated On-Line Isolation and Fractionation Method for Subpopulations of Extracellular Vesicles.

Immunoaffinity chromatography (IAC) with selective antibodies immobilized on polymeric monolithic disk columns enables selective isolation of biomacromolecules from human plasma, while asymmetrical flow field-flow fractionation (AsFlFFF or AF4) can be used for further fractionation of relevant subpopulations of biomacromolecules (e.g., small dense low-density lipoproteins, exomeres, and exosomes) from the isolates. Here we describe how the isolation and fractionation of subpopulations of extracellular vesicles can be achieved without the presence of lipoproteins using on-line coupled IAC-AsFlFFF. With the developed methodology, it is possible to have fast, reliable, and reproducible automated isolation and fractionation of challenging biomacromolecules from human plasma with a high purity and high yields of subpopulations.

Liquid Chromatographic Determination of <i>o</i>-Phosphoethanolamine in Human Plasma Using Fluorescent Derivatization

In this study, an HPLC analysis method using pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) was developed for the determination of o-phosphoethanolamine (PEA), which is a potential biomarker for the diagnosis of major depressive disorder, in human plasma sample. After PEA was derivatized with AQC under mild conditions, the obtained derivative was subjected to purification with a titanium dioxide-modified monolithic silica spin column (MonoSpin® TiO). The eluate from the MonoSpin® TiO was directly injected into an amide-type hydrophilic interaction liquid chromatography (HILIC) column-equipped HPLC system, and the resulting derivative could be separated on the column under alkaline mobile phase conditions and subsequently detected fluorometrically at excitation and emission wavelengths of 250 and 395 nm, respectively. The limit of detection and limit of quantification for a 10 µL injection volume of PEA were 0.052 and 0.17 µM, respectively. The method was validated at 0.2, 1.0, and 5.0 nmol/mL levels in plasma sample, and the precision values were 2.0-6.6% as relative standard deviation and the correlation coefficient (r) of the calibration curve was 0.9995. Furthermore, applicability of this method was demonstrated by analyzing PEA levels in plasma samples from mental illness patients.

Reversed-Phase Liquid Chromatography Methods Based on C18 Polymer Monoliths for the Determination of Urinary Myoglobin

This study describes reversed-phase liquid chromatography (RPLC) methods to quantify urinary myoglobin using polymer monolithic columns produced by copolymerization of stearyl methacrylate (SMA) and ethylene glycol dimethacrylate (EDMA). The columns were prepared in the coffins of 1.5 mm internal diameter (i.d.) ethylene tetrafluoroethylene (ETFE) tubing for use in sequential injection chromatography (SIC) and solid phase extraction (SPE), and inside 1.0 mm i.d. Silcosteel® tube for use in narrow-bore liquid chromatography. The monoliths inside the ETFE were produced via UV, whereas thermal polymerization formed the monoliths inside the Silcosteel® tube. The separation of carbonic anhydrase, lysozyme, and myoglobin was demonstrated because they may occur simultaneously in urine samples. Quantification was undertaken by external calibration, and the accuracy was evaluated by the spiking/recovery strategy. The methods exhibited linearity from 5.0 to 60 µg mL-1 (SIC), 2.5 to 50 µg mL-1 (high-performance liquid chromatography (HPLC)), and 1.0 to 7.5 µg mL-1 for an SPE-HPLC method. The lowest limits of detection and quantifications were 0.13 and 0.43 µg L-1, respectively, obtained after concentrating myoglobin by SPE. Recoveries ranged from 98 to 105%. The low cost, simplicity, reusability, and analytical features provided by these polymeric stationary phases make them affordable alternatives to routine analyses of urinary myoglobin.

Online highly selective recognition of domoic acid by an aptamer@MOFs affinity monolithic column coupled with HPLC for shellfish safety monitoring.

Enabling cost-effective safety monitoring of shellfish is an important measure for the healthy development of the coastal marine economy. Herein, a new aptamer@metal-organic framework (MOF)-functionalized affinity monolithic column was proposed and applied in selective in-tube solid-phase microextraction (IT-SPME) coupled with HPLC for the accurate recognition of domoic acid (DA) in shellfish. Using a surface engineering strategy, ZIF-8 MOF was grown in situ inside the poly(epoxy-MA-co-POSS-MA) hybrid monolith. A high BET surface area and abundant metal reactive sites of the MOF framework were obtained for anchoring massive aptamers with terminal-modified phosphate groups. Various characterizations, such as SEM, elemental mapping, XRD, and BET, were performed, and the affinity performance was also studied. The presence of a massive amount of aptamers with a super coverage density of 3140 μmol L-1 bound on ZIF-8 MOF activated a high-performance bionic-affinity interface, and perfect specificity was exhibited with little interference of tissue matrixes, thus assuring the highly selective capture of DA from the complex matrixes. Under the optimal conditions, DA toxins in shellfish were detected with the limit of detection (LOD) of 7.0 ng mL-1 (equivalent to 14.0 μg kg-1), representing a 5-28 fold enhancement in detection sensitivity over traditional SPE or MIP adsorbents reported previously. The recoveries of fortified mussel and clam samples were achieved as 91.8 ± 1.2%-94.1 ± 1.9% (n = 3) and 91.2 ± 1.1%-94.5 ± 3.6% (n = 3), respectively. This work sheds light on a cost-effective method for online selective IT-SPME and the accurate monitoring of DA toxins using an aptamer@MOF-mediated affinity monolith system coupled with the inexpensive HPLC-UV technique.

Simultaneous Quantitative Determination of Eight Fluoroquinolones by High-Performance Liquid Chromatography with UV Detection using a C18 Monolithic Column

In the present work, an accurate and reliable reversed-phase liquid chromatographic method (RP-HPLC) for the simultaneous quantitative determination of eight fluoroquinolones (enoxacin, norfloxacin, ciprofloxacin, levofloxacin, moxifloxacin, enrofloxacin, sparfloxacin, and marbofloxacin) in bulk form and in pharmaceutical products is described. The optimum separation of the selected pharmaceutical compounds was achieved on a Chromolith® RP-18 endcapped column with a mobile phase consisting of acetonitrile and phosphate buffer (pH 5.5; 15 mM) pumped at 2 mL min-1. UV detection of all compounds was accomplished at 280 nm. The procedure was validated according to the International Conference on Harmonization (ICH) Q2 (R1) guideline. Good linear relationships were obtained (r ≥ 0.996) for all the analytes over their respective concentration ranges. Intra- and inter-day precision were less than 2 %. Recoveries were held in the range of 98 to 102 %. The developed method was successfully applied to the analysis of pharmaceutical products.

Nanoparticle Formulation Composition Analysis by Liquid Chromatography on Reversed-Phase Monolithic Silica.

Multifunctional nanoparticle (NP) formulations for medical purposes have already found their way toward envisaged translation. A persistent challenge of those systems is, next to NP size analysis, the compositional analysis of the NPs with the polymer as the matrix component and the encapsulated drug, particularly in a quantitative manner. Herein, we report the formulation of poly(lactic-co-glycolic acid) (PLGA) NPs by nanoprecipitation and the analysis of their integrity and size by dynamic light scattering (DLS) and scanning electron microscopy (SEM). Those NPs feature a variety of encapsulated drugs including the well-known ibuprofen (Ibu) as well as dexamethasone (Dex) and dexamethasone acetate (DexAce), with the latter being of potential interest for clinical treatment of SARS-CoV-2 patients. All those dissolved formulation compositions have been subjected to liquid chromatography on reversed-phase silica monolithic columns, allowing to quantitatively assess amounts of small molecule drug and NP constituting PLGA polymer in a single run. The chromatographically resolved hydrophobicity differences of the drugs correlated with their formulation loading and were clearly separated from the PLGA matrix polymer with high resolution. Our study identifies the viability of reversed-phase monolithic silica in the chromatography of both small drug molecules and particularly pharmapolymers in a repeatable and simultaneous fashion, and can provide a valuable strategy for analysis of diverse precursor polymer systems and drug components in multifunctional drug formulations.

Influence of ion-pairing reagents on the separation of intact glycoproteins using hydrophilic-interaction liquid chromatography - high-resolution mass spectrometry

Hydrophilic-interaction liquid chromatography (HILIC) of intact proteins offers high-resolution separations of glycoforms of glycoproteins differing in the number of (neutral) glycans. However, to obtain efficient separations it is essential that the positively charged sites of the proteins are shielded by acidic (negative) ion-pair reagents (IPRs), so as to enhance the contribution of the hydroxyl groups of the (neutral) sugars in the glycoprotein. Here, we studied the influence of various IPRs that differ in physico-chemical properties, such as hydrophobicity and acidity, on the capillary-scale HILIC separation of intact (glyco)proteins. We evaluated the use of fluoroacetic acid (MFA), difluoroacetic acid (DFA), trifluoroacetic acid (TFA), and heptafluorobutyric acid (HFBA) as diluents for sample preparation, as solvents for sample loading on a reversed-phase trap prior to the HILIC separation, and as mobile-phase components for HILIC and HILIC-MS. To reduce the contribution of ion-exchange interaction with the (silica-based) stationary phase, we used an acrylamide-based monolithic column. We studied the influence of the different IPRs on each step of the separation of a mixture of proteins of different size and hydrophilicity and on the separation of the five glycoforms of ribonuclease B. The content of IPR in the sample was shown not to affect the separation and the MS detection. However, a low content of TFA and DFA in the mobile phase is favourable, as it reduces adduct formation and leads to higher signal intensity. The optimized HILIC conditions successfully resolved nine major glycoforms groups of a ∼40 kDa glycoprotein horseradish peroxidase (HRP), as an example of a complex glycoprotein.

Synthesis and characterization of supermacroporous cryogel with immobilized p-aminobenzenesulfonamide as affinity ligand for the purification of lactoperoxidase from whey.

This study proposed the development of a monolithic supermacroporous affinity column for direct capture of lactoperoxidase, a glycoprotein present in milk, whey, and colostrum, with several applications due to its wide antimicrobial activity. A poly(acrylamide)-based cryogel was produced by radical co-polymerization of monomers in frozen aqueous solution and activated with p-aminobenzenesulfonamide as a ligand for specific interaction with the lactoperoxidase. The axial liquid dispersion coefficients at different liquid flow rates were determined by measuring residence time distributions using the tracer pulse-response method. The axial dispersion coefficient was low and the height equivalent to theoretical plate was not dependent on the flow velocity. The adsorptive capacity of affinity cryogel was studied as a function of flow velocity and the best condition was 0.9cm/min. The response surface methodology was applied to optimize the capture of the enzyme, as a function of pH and salt concentration. Higher purification factor value was found at a salt concentration of 80mmol/L and pH of 8.0 (p<0.05). There was no influence of the variables under study on the yield (p>0.05). The results indicated that affinity cryogel is a promising chromatography support for the use in high-throughput one-step purification of lactoperoxidase from whey.

Covalent organic framework-based monolithic column with hydrophilic and π-π stacking interaction for efficient in-tube solid-phase microextraction of synthetic phenolic antioxidants

Currently, covalent organic framework-based (COF-based) monolithic columns show excellent application potential for high-efficient sorbent in sample preparation. Hydrophobic and π-π stacking interaction were their primary adsorption mechanisms for extraction. Hydrophilic interaction has not been involved yet. Herein, a novel COF-based monolithic column with hydrophilic interaction and π-π stacking interaction has been fabricated by the embedment of COFTAPB-TPA microspheres into urea–formaldehyde resin (UF) monolith. Due to the excellent adhesion performance of UF, the COFTAPB-TPA microspheres could be steadily embedded into the UF monolith. Taking the advantages of the hydrophilic interaction and the π-π stacking interaction between the COFTAPB-TPA@UF monolith and synthetic phenolic antioxidants (SPAs), the COFTAPB-TPA@UF monolith was adopted as efficient sorbent for in-tube SPME of trace SPAs in edible oils. Several SPME parameters, including the percentage of ACN in the sampling solution and eluent, the collection time span, and the pH value of the sampling solution, were optimized for satisfactory SPME performance. Under the optimized conditions, the LODs of the analytes varied from 0.2 to 1.2 ng/mL, and the recoveries ranged from 78.1 % to 98.4 % for sunflower oil samples at three spiked levels, with the RSDs lower than 7.2 %, indicating the superior extraction capacity of the COFTAPB-TPA@UF monolith as well as the promising feasibility of the proposed in-tube SPME-HPLC method. This work lighted a facile way for COF-based monoliths with hydrophilic interaction, and further expanded their applications for sample preparation.

Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples.

Protein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multianalyte methods allowing the establishment of a uremic metabolite profile are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fluorescence detection for the simultaneous quantification of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate buffer containing an ion-pairing reagent, at a flow rate of 2mLmin-1. Standards were prepared in plasma and quantification was performed using the background subtraction approach. LOQ values were ≤ 0.2µgmL-1 for all analytes except for pCS (LOQ of 2µgmL-1). The method proved to be accurate (93.5-112%) and precise (CV ≤ 14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting significantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression.

N, N′-methylene bisacrylamide/divinyl benzene based-highly cross-linked hybrid monolithic column: Production and its applications for powerful capture of four chlorophenols

In this paper, the role of the halogen bond in capillary monolithic column microextraction was explored for the first time. Benzene-1,3,5-tricarbohydrazide (BTH) was synthesized as a functional monomer, N, N′-methylene bisacrylamide (MBA) and divinyl benzene (DVB) were used as cross-linking agents, the hybrid monolithic column of poly (BTH-co-DVB-co-MBA) was prepared using methanol and polyethylene glycol as pore-forming agents and azodiisobutyronitrile as the initiator. Due to the existence of BTH, a large number of nitrogen atoms (Lewis base) were introduced into the monolithic column, which could form a halogen bond with chlorine-containing organic pollutants and enhance its adsorption performance. Based on the monolithic column, a sensitive and environment-friendly solid-phase microextraction technology was studied. The monolithic column was integrated with high-performance liquid chromatography (HPLC) to extract and detect four kinds of chlorophenol in real water samples. Under best conditions, the method showed excellent extraction ability and linearity, with a linear correlation coefficient of 0.9958–0.9987, a low detection limit (LOD) of 0.04–0.23 μg L−1 (S/N = 3), and relative standard deviation (RSD) less than 3.09%. The recovery rate was kept between 87.30% and 123.00%, and the RSD was less than 3.83%, which indicated that the column had powerful capture performance, high precision, and strong anti-matrix interference ability in the real sample, and had potential application value in practical work.