Preparation of spherical silica hydroxyl-functionalized covalent organic polymer composites for mixed-mode liquid chromatography.

Covalent organic polymers are an emerging class of amorphous microporous materials that have raised increasing concerns in analytical chemistry due to their unique structural and surface chemical properties. However, the application of covalent organic polymers as mixed-mode stationary phases in chromatographic separations has rarely been reported. Herein, novel spherical silica hydroxyl-functionalized covalent organic polymer composites were successfully prepared via a layer-by-layer approach. The structure and morphology of the materials were carefully characterized by elemental analysis, Fourier-transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, X-ray diffraction, thermogravimetric analysis, Brunauer-Emmett-Teller, and contact angle measurements. Baseline separations of various alkylbenzenes, polycyclic aromatic hydrocarbons, and nucleosides and bases were achieved on the prepared stationary phase under reversed-phase/hydrophilic interaction mode. The column efficiencies of 23 853 and 36 580 plates/m were obtained for butylbenzene and uracil, respectively, and the relative standard deviation of the retention time for continuous injections was less than 1.38% (n = 10), suggesting satisfactory column efficiency and repeatability. Additionally, this novel stationary phase realized the complete separation of the endocrine-disrupting chemicals in river water. This work affords a new route for synthesizing covalent organic polymers-based mixed-mode stationary phase and further reveals their great potential in chromatographic separation.

A ratiometric fluorescence sensor for ascorbic acid determination based on an AND-NAND logic pair.

An AND-NAND logic pair is reported based on non-purified carbon quantum dots (CDs) for ascorbic acid (AA) detection. In the logic operation, molybdenum oxide nanosheets (MoO3 NSs) and AA are used as two signal inputs. In the presence of AA, MoO3 NSs are reduced to plasmonic molybdenum oxide, which decreases the CD fluorescence intensity because of a static quenching, dynamic quenching, and internal filtration effect. Meanwhile, the AA is oxidized to dehydroascorbic acid and forms fluorescent 3-(dihydroxyethyl) furo [3,4-b] quinoxaline-1-one with o-phenylenediamine from non-purified CDs. On this basis, an AND-NAND logic pair was constructed and used as a ratiometric fluorescence sensor for highly sensitive detection of AA. The method has a wide linear range of 0.05-50 µM, and a detection limit of 34 nM. In addition, it was used to detect AA in fresh fruit. Potential applications include chemical computing, optoelectronic devices, biomedical science, and environmental monitoring. HIGHLIGHTS: 1. A ratiometric fluorescence sensor based on AND-NAND logic pair constructed by CDs and MoO3 NSs was successfully fabricated. 2. The ratiometric fluorescence sensor exhibited satisfactory linear range, high sensitivity, and good selectivity for AA. 3. The ratiometric fluorescence method was able to detect AA in fresh fruit with good results comparable to official fluorescence methods.

Simultaneous determination of cytokinins by high performance liquid chromatography with resonance Rayleigh scattering and mechanism discussion.

A reliable, highly sensitive and highly selective method of high performance liquid chromatography associated with resonance Rayleigh scattering (HPLC-RRS) was developed to detect three cytokinins, namely, 6-benzylaminopurine (BA), kinetin (KT) and zeatin (ZT). In this work, Pd(ii) is added into the system to form ternary ion association complexes for the first time, which results in a lower limit of detection and extends the application of HPLC-RRS. The experimental conditions were optimized. In order to investigate the reaction mechanism, the ternary ion association complexes were characterized by ultraviolet-visible spectrophotometry, dynamic light scattering, scanning electron microscopy and density functional theory calculations. In a HAc-NaAc buffer solution (pH = 4.1), a ternary complex of cytokinin : Pd(ii) : EryB (1 : 1 : 2) was formed. The detection limits (S/N = 3) of BA, KT, and ZT were 0.9, 1.5 and 2.3 ng mL-1, respectively. In addition, this method was applied for the simultaneous detection of cytokinins in real samples with satisfactory results.

A quadruple-channel fluorescent sensor array based on label-free carbon dots for sensitive detection of tetracyclines.

A quadruple-channel fluorescent sensor array based on label-free carbon dots (CDs) was fabricated to detect and discriminate a series of tetracyclines (TCs), including chlortetracycline (CTC), oxytetracycline (OTC), tetracycline (TC) and doxycycline (DOX). Blue-emitting carbon dots (B-CDs) and green-emitting carbon dots (G-CDs) were prepared to serve as four sensing elements. When the TCs were directly mixed with CDs, the fluorescence quenching phenomenon appeared. Since different TCs exhibited different affinities for sensing elements, the sensor array displays a distinct fluorescence pattern of the fluorescence intensity variation (F0 - F)/F0 for each of these TCs, which is further analyzed by principal component analysis (PCA). The present fluorescent sensor array has the capacity to differentiate TCs at a low concentration of 1 µM. Meanwhile, quantitative detection with a lower limit (0.30 µM) for TCs could be achieved by applying a single element. Moreover, a high accuracy (100%) examination of unknown samples is acquired. Finally, the fluorescent sensor array performs well in distinguishing binary mixtures and could also recognize TCs in milk.

One-step synthesis of N-doped carbon dots, and their applications in curcumin sensing, fluorescent inks, and super-resolution nanoscopy.

Nitrogen-doped carbon dots (N-CDs) with fluorescence excitation/emission maxima at 365/450 nm were prepared by a one-step hydrothermal approach. The dots possess remarkable photostability, fluorescence blinking and good biocompatibility, and this favors utilization in stochastic optical reconstruction microscopy (STORM). A spatial resolution down to ~60 nm was achieved when imaging HeLa cells using 647-nm laser excitation. This opens new possibilities for various super-resolution techniques based on stochastic optical switching. The remarkable optical properties of the N-CDs also enable them to be applied as invisible security ink for use in patterning, information storage and anti-counterfeiting. Further, it is found that the fluorescence of the N-CDs can be quenched by curcumin with high efficiency due to a combination of inner filter effect and static quenching. Based on this, a fluorometric assay with a detection limit of 21 ng mL-1 was developed for the determination of curcumin. Graphical abstract Schematic representation of the applications of N-doped carbon dots (N-CDs). Curcumin quenches the fluorescence of N-CDs with high efficiency. The remarkable optical properties of the N-CDs enable them to be applied in fluorescent ink, cell imaging and stochastic optical reconstruction microscopy (STORM).

Preparation and evaluation of surface-bonded phenylglycine zwitterionic stationary phase.

4-Hydroxy-D-phenylglycine was modified with methacrylic anhydride and then immobilized on silica through thiol-initiated surface polymerization; the prepared material was applied as stationary phase for HPLC. The obtained stationary phase was characterized by elemental analysis, infrared spectroscopy, and thermogravimetric analysis. The chromatographic performance of the packed column was evaluated in reversed-phase liquid chromatograph (RPLC) and hydrophilic interaction liquid chromatograph (HILIC) mode; this column has shown excellent selectivity to both the hydrophobic and hydrophilic solutes. The selectivity towards polycyclic aromatic hydrocarbons relative to that towards alkylbenzenes exhibited by the prepared column was higher than the corresponding selectivity exhibited by commercial C18 column, which could be explained by electronic π-π interaction between phenylglycine and electron-rich aromatic rings. On the other hand, the prepared column has also shown better selectivity for polar compounds, which was based on the multiple interaction and retention mechanisms. It was also used to separate sulfonamides and organic acid compared with a commercial C18 and HILIC column; the results show its great chromatographic performance with distinctive selectivity. All the results indicated the prepared column had potential application in a wide range.

High performance liquid chromatography associated with resonance Rayleigh scattering for synchronous determination of three antihistamines and mechanism study.

A highly sensitive and selective method of high performance liquid chromatography (HPLC) combined with resonance Rayleigh scattering (RRS) spectra was developed for the detection of three antihistamine drugs, including pyrilamine (PY), carbinoxamine (CAR) and tripelennamine (TRI). The three antihistamines were separated by a C18 column. The mobile phase contained 25% acetonitrile (ACN) and 75% phosphate buffered solution (pH 3.2) with the flow rate of 0.4 ml min-1 . In medium of Britton-Robinson (BR) buffer solution (pH 4.6), the PY, CAR and TRI separated by HPLC and then reacted with Erythrosine B (EryB), forming 1:1 ion-association complexes, which led to significant signal enhancement of RRS spectra. The RRS spectra was detected at the wavelength λex =λem = 370 nm. The calibration curves of PY, CAR and TRI were linear in the range from 0.02 to 25 µg ml-1 , and the detection limit [signal-to-noise ratio (S/N) = 3] were 3.38, 4.48 and 5.50 ng ml-1 , respectively. In addition, under the optimum experiment condition, the reaction mechanism and the reasons for RRS enhancement were investigated in this work. The developed method was applied to the simultaneous detection of three antihistamines in water samples with satisfying results.

High-performance liquid chromatography study of gatifloxacin and sparfloxacin using erythrosine as post-column resonance Rayleigh scattering reagent and mechanism study.

Herein, a highly selective high-performance liquid chromatography (HPLC) coupled with resonance Rayleigh scattering (RRS) method was developed to detect gatifloxacin (GFLX) and sparfloxacin (SPLX). GFLX and SPLX were first separated by HPLC, then, in pH 4.4 Britton-Robinson (BR) buffer medium, protonic quaternary ammonia cation of GFLX and SPLX reacted with erythrosine (ERY) to form 1:1 ion-association complexes, which resulted in a significant enhancement of RRS signal. The experimental conditions of HPLC and post-column RRS have been investigated, including detection wavelength, flow rate, pH, reacting tube length and reaction temperature. Reaction mechanism were studied in detail by calculating the distribution fraction. The maximum RRS signals for GFLX and SPLX were recorded at λex  = λem  = 330 nm. The detection limits were 3.8 ng ml-1 for GFLX and 17.5 ng ml-1 for SPLX at a signal-to-noise ratio of 3. The developed method was successfully applied to the determination of GFLX and SPLX in water samples. Recoveries from spiked water samples were 97.56-98.85%.

Resonance Rayleigh scattering technique as a detection method for the RP-HPLC determination of local anaesthetics in human urine.

A highly selective and sensitive method of reversed phase high-performance liquid chromatography (RP-HPLC) coupled with resonance Rayleigh scattering (RRS) was developed for the determination of procaine, bupivacaine and tetracaine. Separation of three local anaesthetics was achieved at 35 °C on a C18 column. The mobile phase was 30: 70 (v/v) acetonitrile/triethylamine-phosphoric acid buffer (pH 2.9) at flow rate of 0.3 mL/min. The RRS detection was conducted by taking advantage of the strong RRS enhancement of the local anaesthetics with erythrosine reaction in an acidic medium. Under optimum conditions, the limit of detection (S/N = 3) values were in the range of 2.4-11.2 ng/mL. Recoveries from spiked human urine samples were 95.8%-104.5%. The proposed method applied to the determination of local anaesthetics in human urine achieved satisfactory results. In addition, the mechanism of the reaction is fully discussed. Copyright © 2016 John Wiley & Sons, Ltd.

Study on the interaction between three benzimidazole anthelmintics and eosin Y by high performance liquid chromatography associating with resonance light scattering and its application.

A novel, highly selective, and sensitive resonance light scattering (RLS) detection approach coupled with high performance liquid chromatography (HPLC) was researched and developed for the synchronous analysis of three kinds of benzimidazole anthelmintics, including mebendazole (MBZ), albendazole (ABZ), and fenbendazole (FBZ) for the first time. In the pH range of 3.5-3.7 Britton-Robinson buffer medium, three kinds of anthelmintics, which were separated by HPLC, reacted with eosin Y (EY) to form 1:1 ion-association complexes, resulting in significantly enhanced RLS signals and the maximum peak located at 335 nm. The enhanced RLS intensity was in proportion to the MBZ, ABZ, and FBZ concentration in the range 0.2-25, 0.2-23, and 0.15-20 µg/mL, respectively. The limit of detection was in the range of 0.064-0.16 µg/mL. In addition, human urine was determined to validate the proposed method by spiked samples and real urine samples. Satisfactory results were obtained by HPLC-RLS method. Graphical Abstract The diagram mechanism of generating resonance between emitted light and scattered light.

Fabrication of ionic liquid functionalized silica with different anions and the application in mixed-mode and chiral chromatography.

To realize the potential of ionic liquid functionalized silica to prepare mixed-mode and chiral stationary phases, two ionic liquid silane reagents with different anions were synthesized via a high-efficiency click reaction. Then they were decorated onto the surface of silica by a one-step bonding reaction. The functionalized silica was characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and elemental analysis (EA). Two stationary phases provided satisfactory performance when compared with a commercial mixed-mode column. Notably, Sil-C10Im-D-BCS with D-3-bromocamphor-8-sulfonate (D-BCS) as anion presented chiral separation capacity towards 1,2,3,4-Tetrahydro-1-naphthol. The separation mechanism was investigated through multiple pathways, and the results revealed that the prepared stationary phases can retain and separate solutes through multiple interactions, like hydrophobic effect, ion exchange, hydrogen-bond interaction, etc. Quantum chemical calculation (QC) was employed to obtain the optimized structures and the binding energy of anions to cations. The results provided some insights into the retention mechanism from a molecular perspective. This work demonstrated the superiority of ionic liquid functionalized silica as mixed-mode stationary phases and the potential of chiral ionic liquid as chiral selectors.

Preparation of two zwitterionic polymer functionalized stationary phases and comparative evaluation under mixed-mode of reversed phase/ hydrophilic interaction/ion exchange chromatography.

Zwitterions are a promising choice to prepare separation materials because of their hydrophilicity and biocompatibility. We described the preparation of two zwitterionic polymer functionalized stationary phases and evaluation under mixed-mode chromatography. A zwitterionic monomer, S-(4-vinylbenzyl) cysteine (SVC), was synthesized and bonded to silica via reversible addition fragmentation chain transfer (RAFT) polymerization to afford a zwitterionic stationary phase, Sil-SVC. A hydrophobic monomer, N-(4-phenylbutan-2-yl) acrylamide (NPA), was copolymerized with SVC onto the stationary phase (Sil-SVCNPA) for comparison. The stationary phases were characterized with FT-IR, TGA, EA, and zeta-potential measurements. Mobile phase composition (ACN content, pH and salt concentration) was varied to study the retention property. Linear solvation energy relationship and Van't Hoff plot were used to investigate the retention mechanism and how chromatographic conditions influenced it. Both stationary phases showed a mixed-mode of RPLC/HILIC/IEC and satisfactory performance in separating hydrophobic analytes (alkylbenzenes and polycyclic aromatic hydrocarbons), hydrophilic nucleotide and bases, and anions, high column efficiency of 60,000 plates·m-1 was achieved. In summary, zwitterionic polymers are attractive options to prepare stationary phases and the retention property can be easily regulated by copolymer.

Dicationic imidazole ionic liquid stationary phase for preservative detection and its application under mixed mode of HILIC/RPLC/IEC.

BACKGROUND: Food safety has always been a great concern, and the detection of additives is vital to ensuring food safety. Therefore, there is a necessity to develop a method that can quickly and efficiently separate and detect additives in food. High performance liquid chromatography is widely used in the analysis and testing of food additives. Ionic liquids have attracted wide attention in the preparation of high performance liquid chromatography stationary phases owing to their high stability, low vapor pressure and adjustable structure. RESULTS: We developed a novel dicationic imidazole ionic liquid stationary phase for the simultaneous determination of organic preservatives (sodium benzoate, potassium sorbate) and inorganic preservatives (nitrate and nitrite) in foodstuffs under mixed-mode chromatography. The method had the advantages of easy operation, high reproducibility, good linearity and precision. In the detection of these four preservatives, the limit of detection ≤0.4740 mg⋅L-1 and the limit of quantification ≤1.5800 mg⋅L-1. The intra-day and inter-day precision were less than 4.02%, and the recovery rate was 95.90∼100.19 %. At the same time, we also characterized the stationary phase, explored the mechanism and evaluated the chromatographic performance. The stationary phase was able to operate under the mixed mode of reversed phase/hydrophilic interaction/ion exchange chromatography, and it was capable of separating hydrophilic substances, hydrophobic substances, acids, and inorganic anionic substances with good separation efficiency and had high column efficiency. SIGNIFICANCE: In summary, the stationary phase has a promising application in the routine analysis of organic and inorganic preservatives in food. In addition, the stationary phase has good separation ability for hydrophilic, hydrophobic, ionic substances and complex samples, making it a prospective material for chromatographic separation.

An aptasensing platform for simultaneous detection of multiple analytes based on the amplification of exonuclease-catalyzed target recycling and DNA concatemers.

In the present study, an ultrasensitive electrochemical aptasensor for the simultaneous detection of thrombin (TB) and ochratoxin A (OTA) was fabricated by using exonuclease-catalyzed target recycling and DNA concatemers for signal amplification. The previously hybridized double-stranded DNAs (SH-cTBA/TBA and SH-cOBA/OBA) were self-assembled on a gold electrode. In the presence of targets, the formation of aptamer-target complexes would lead to not only the dissociation of aptamers (TBA and OBA) from the double-stranded DNAs but also the transformation of the complementary DNAs (SH-cTBA and SH-cOBA) into hairpin structures. Subsequently, owing to employment of RecJf exonuclease, which is a single-stranded DNA-specific exonuclease to selectively digest the appointed DNAs (TBA and OBA), the targets could be liberated from the aptamer-target complexes for recycling of the analytes. Thereafter, probe DNAs (T1 and T2) were employed to hybridize with SH-cTBA and SH-cOBA respectively to provide primers for the concatemer reaction. After that, when four auxiliary DNA strands S1, anthraquinone-2-carboxylic acid (AQ)-labeled S2, S3, S4, as well as hemin were introduced, extended dsDNA polymers with lots of AQ moieties and hemin-G-quadruplex complexes could form on the electrode surface. Then, based on the signal of the AQ and hemin-G-quadruplex complex, an electrochemical aptasensor for the simultaneous detection of TB and OTA was successfully fabricated.

Preparation of three regioisomeric ionic liquid stationary phases and investigation of their retention behavior.

The structural properties of ionic liquid stationary phases have a considerable effect on their separation selectivity. However, the difference of the chromatographic retention behavior of different regioisomeric ionic liquid stationary phases has rarely been investigated. In this study, three regioisomeric ionic liquid silane reagents were prepared by photoinitiated ene-click chemistry and bonded to silica by one-pot method to fabricate three new stationary phases (Sil-C2Im-C8, Sil-C6Im-C4, and Sil-C9Im-C1). All three stationary phases showed promising retention repeatability and efficiency. The retention behavior of the three stationary phases was investigated under various chromatographic conditions. The retention mechanism was further investigated by the linear energy solvation relationship and Van't Hoff plots. The stationary phases exhibited mixed-mode retention mechanisms. The π-π, hydrogen bonding, ion-exchange, and hydrophilic interactions with analytes were the weakest when the imidazole ions were embedded in the innermost part of the alkyl chains, while the interactions were the strongest when the imidazole ions were embedded in the middle of the alkyl chains. The three stationary phases provided great but different separation performances towards nucleosides, nucleobases, aromatic acids, alkyl benzenes, and polycyclic aromatic hydrocarbons due to the influence of imidazole ion position.

Preparation of three structurally similar stationary phases with different ionizable terminal groups and evaluation of their retention performances under multiple modes in high performance liquid chromatography.

Three structurally similar silane reagents with different terminal groups were prepared and bonded to silica to obtain three structurally similar stationary phases (Sil-Ph-COOH, Sil-Phe and Sil-Ph-NH2). The prepared stationary phases were characterized through elemental analysis (EA) and Fourier Transform Infrared Spectroscopy (FT-IR). These three stationary phases provided acceptable retention repeatability (relative standard deviations between 0.08% and 0.13%) and high column efficiency (7.3 × 104 plates/m for uridine on Sil-Phe). The retention behavior of the three columns was investigated under different chromatographic conditions including different mobile phase ratio, salt concentration, pH etc. The retention mechanisms were explored by linear solvation energy relationships and Van't Hoff plots. Applications in separation under reversed phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) mode were investigated. The results showed that the retention capacity of the stationary phases with different terminal groups to the analytes is very different, especially for carboxylic acids, because the surface charges of amino groups and carboxyl groups under weakly acidic conditions produce different electrostatic effects with dissociated carboxylic acids. Finally, the Sil-Phe column was employed to detect ibuprofen extracted from pharmaceutical ibuprofen capsules and vitamins extracted from vitamin tablets.

Effect of spacer alkyl chain length on retention among three imidazolium stationary phases under various modes in high performance liquid chromatography.

The properties of ionic liquids (ILs) greatly influenced the retention of solutes on IL-functionalized stationary phases. In this work, three IL-functionalized stationary phases (Sil-C4Im, Sil-C7Im, and Sil-C10Im) were prepared by modifying silica gel with three ionic liquid silane reagents differing in spacer alkyl chain lengths, which refers to the alkyl chain between the imidazolium and the sulfur atom. The preparation was proved through a range of characterization techniques, including elemental analysis (EA), Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). The effects of chromatographic conditions, such as acetonitrile content, salt concentration, pH, and column temperature, were studied to explore the retention mechanism of three stationary phases, which indicates that three IL-stationary phases provide interactions with solutes in reverse-phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC), and ion exchange chromatography (IEC). The column efficiency is 83,686 plates/m for aminobenzoic acid, and the column also has excellent repeatability of retention time with relative standard deviations (RSDs) between 0.24% and 0.34% (n = 12). A variety of analytes, including five nucleosides and nucleobases, six alkylbenzenes, two polycyclic aromatic hydrocarbons (PAHs), and four inorganic anions, were separated well on three IL-functionalized stationary phases. It was found that the spacer alkyl chain length influenced selectivity by comparing the retention of the three IL-functionalized stationary phases.

Solvent-dependent carbon dots for multifunctional sensing of temperature, pH, and proton pump inhibitors.

Multifunctional sensing, a strategy to improve the efficiency of analysis and detection, has attracted much attention. In this study, a multifunction sensing platform was developed for temperature, pH, and proton pump inhibitors (PPI) with fluorescence carbon dots. Solvent-dependent carbon dots (PG-CDs) were synthesized through a one-step solvothermal method from phloroglucinol with the assistance of HCl. Base on the effect of functional groups on the surface, solvent-dependent behavior, temperature and pH responsive fluorescence can be observed. The PG-CDs display a reversible temperature-sensitive fluorescent behavior within 15-65 °C in N, N-dimethylacetamide. The fluorescence intensity of PG-CDs also responded to pH in a range of 3.0-7.0 with good reversibility and anti-interference. Therefore, the platform for temperature and pH sensing was proposed. In addition, since the synergistic effect of dynamic and static quenching, proton pump inhibitors (PPI) can be detected sensitively, including esomeprazole (EPZ), omeprazole (OPZ), and rabeprazole (RPZ). The linear range of detection for EPZ, OPZ, and RPZ is 2.0-80.0, 2.0-75.0, and 10.0-200.0 µM and the limits of detection is 0.29, 0.55, and 2.99 µM, separately. The recoveries for real samples were between 91.83 and 102.3%, which indicated that the platform for PPI sensing had good accuracy.

The dynamics of a novel method coupling high-performance liquid chromatography with resonance Rayleigh scattering.

The dynamics of a novel method coupling high-performance liquid chromatography (HPLC) with resonance Rayleigh scattering (RRS) is presented in the paper. The method was employed in the detection of fluoroquinolones (FQs) of urine samples from healthy human beings, by forming ion-association complexes between the components separated from HPLC and [Ce(OH)(3) ](+) as the molecular recognition probe. The RRS signal was measured at λ(ex) =λ(em) =365 nm. It was applied to detect three FQs and obtained satisfactory results. The RRS spectral characteristics of the analytes and the kinetics of flow rate, proportion of organic phase, reaction time and the aggregation level of ion-association complexes were investigated, which provided a new basis for the development of the hyphenated techniques. It was established that the presence of HPLC-RRS would open up a new field in the determination of analytes in the absence of UV or fluorescence.

Facile fabrication of silica@covalent organic polymers core-shell composites as the mixed-mode stationary phase for hydrophilic interaction/reversed-phase/ion-exchange chromatography.

Covalent organic polymers (COPs) are a promising class of cross-linked polymeric networks that attracted extensive attention in separation and analysis fields. Exploring facile and convenient strategy to prepare COPs-based mixed-mode stationary phases for high performance liquid chromatography (HPLC) has seriously lagged and has never been reported. Herein, we describe a facile in-situ grow strategy for fabrication of silica@COPs core-shell composites (SiO2@TpBD-(OH)2) as a novel mixed-mode stationary phase for HPLC. Owing to the co-existing of abundant hydroxyl, carbonyl, imine, cyclohexyl groups, and benzene rings in the skeleton of COPs shell, the developed mixed-mode stationary phase exhibits hydrophilic interaction liquid chromatography (HILIC)/reversed-phase liquid chromatography (RPLC)/ion-exchange chromatography (IEX) retention mechanisms. The content of acetonitrile, pH value, and salt concentration in the mobile phase were investigated on SiO2@TpBD-(OH)2 packed column. In comparison to conventional single-mode columns, the SiO2@TpBD-(OH)2 column showed flexible selectivity, enhanced separation performance, and superior resolution for benzene homologues, polycyclic aromatic hydrocarbons, nucleosides and bases, and acidic organic compounds. The column efficiency of p-nitrobenzoic acid was up to 54440 plates per meter. The packed column also possessed outstanding chromatographic repeatability for six nucleosides and bases with the RSDs of 0.07-0.23%, 0.58-1.77%, and 0.31-1.23% for retention time, peak area, and peak height, respectively. Besides, the SiO2@TpBD-(OH)2 column offered baseline separation of multiple organic pollutants in lake water, which verified its great potential in real sample analysis. Overall, the silica@COPs core-shell composites not only provide a new candidate of mixed-mode stationary phases, but also extend the potential application of COPs in separation science.