Functional Three-Dimensional Zeolitic Imidazolate Framework with an Ordered Macroporous Structure for the Isolation of Extracellular Vesicles.

Extracellular vesicles (EVs) and their cargoes are increasingly being recognized as noninvasive diagnostic markers, necessitating the isolation of EVs from complex biological samples. Herein, a distearoyl phospholipid ethanolamine-functionalized single-crystal ordered macroporous three-dimensional zeolitic imidazolate framework (SOM-ZIF-8-DSPE) was developed, which combines the surface charge interaction of ZIF-8 with the synergistic effect of DSPE insertion into the phospholipid membrane of EVs to improve the isolating selectivity of EV capture. The materials have porous structures larger than 300 nm in diameter, providing enough space and active sites to trap EVs. Benefiting from this feature, the entire isolation process takes only 10 min and is well compatible with the subsequent analysis of RNA in EVs. Consequently, 10 upregulated miRNA of plasma EVs in the primary colorectal cancer (pCRC) patients is found over the healthy donors, and 6 upregulated miRNA of plasma EVs in the metastatic colorectal cancer (mCRC) patients over pCRC patients. These findings suggest that the isolation of EV-based SOM-ZIF-8-DSPE is a promising strategy to identify biomarkers for disease diagnosis, such as miRNAs in plasma EVs for the early detection of CRC.

External surface hydrophilic 3D-POSS-COF@GDL for the direct adsorption of bisphenols in milk samples.

In case of organic frameworks (COFs) as adsorbents in the pretreatment of complex food matrices, challenges such as poor dispersion and non-specific adsorption of interfering macromolecules like proteins are often encountered. To address this issue, this work prepared a three-dimensional covalent organic framework (3D-COF) with a novel bcu topology based on polyhedral oligomeric silsesquioxane (POSS). Subsequently, gluconolactone (GDL) was modified onto the external surface of the material via the reaction with the exposed reactive residues. The resulting POSS-COF@GDL adsorbent has an enhanced hydrophilicity in the external surface, thereby significantly improves the dispersion of materials in aqueous solution and reduces the adsorption ability toward protein. Whereas, the inner of material retains hydrophobic pores that exhibit high adsorption efficiency to small hydrophobic molecules. Compared with the traditional pretreatment methods, POSS-COF@GDL can directly extract bisphenols (BPs) in milk samples without any additional treatment. The established sample pretreatment method is coupled with high-performance liquid chromatography-ultraviolet detection (HPLC-UV), resulting in recoveries of 71.8 to 93.6%, intra- and inter-day relative standard deviations (RSDs) of <8.3%, and limits of detection (LODs) of 0.042-0.16 ng∙mL-1 for four BPs.

Nitrogen-doped magnetic porous carbon nanospheres derived from dual templates-induced mesoporous polydopamine coated Fe3O4 for efficient extraction and sensitive determination of volatile nitrosamines by gas chromatography-mass spectroscopy.

N-nitrosamines (NAs) are highly carcinogenic compounds commonly found in food, beverages, and consumer products. Due to their wide polarity range, it is challenging to find a suitable carbon adsorbent that can simultaneously adsorb and enrich both polar and nonpolar NAs with good recovery. In this study, nitrogen-doped magnetic mesoporous carbon nanospheres (M-MCN) were prepared and employed as an adsorbent for magnetic solid-phase extraction (MSPE) to extract and concentrate four NAs. The introduction of nitrogen functional groups enhanced the hydrophilicity of the carbon material, allowing M-MCN to achieve a balance between hydrophilicity and hydrophobicity, resulting in good recovery for both polar and nonpolar NAs. A method combining MSPE with gas chromatography-mass spectrometry (GC-MS) was developed for the determination of NAs in processed meat and alcoholic beverages. The method exhibited a good linear range (1-100 ng g-1, r2 > 0.9967) and trace-level detection (0.53-6.6 ng g-1). The recovery rates for the four NAs ranged between 85.7 and 110.7 %, with intra-day precision expressed as relative standard deviation (RSD) between 4.1 and 10.7 %, and inter-day precision between 4.8 and 12.9 %. The results demonstrated not only good accuracy and precision but also provided a new adsorbent for the enrichment of trace-level NAs in processed meat and alcoholic beverage samples.

Facile construction of a stable core-shell spherically magnetic polyimide covalent organic framework for efficient extraction of phenylurea herbicides.

Efficient enrichment is crucial for the highly sensitive monitoring of phenylurea herbicides (PUHs) in various environmental waters. In this work, a stable core-shell spherically magnetic polyimide covalent organic framework (COF) was synthesized via a simple template-mediated precipitation polymerization method under mild conditions using tri(4-aminophenyl)amine (TAPA) and 3,3',4,4'-biphenyltetracarboxylic dianhydride (BPDA) as the building units (denoted as Fe3O4@TAPA-BPDA). The Fe3O4@TAPA-BPDA exhibits remarkable adsorption performance for PUHs with an optimized adsorption time of only 10 min. The adsorption of PUHs by Fe3O4@TAPA-BPDA followed the pseudo-second-order kinetic model and the Langmuir model. Furthermore, hydrogen bonding, halogen bonding, hydrophobic interaction, electro donor-acceptor interaction and π-π interactions are identified as the dominant mechanisms contributing to excellent adsorption performance. It was demonstrated that halogen bonds play an important role in the adsorption of substances containing chlorine atoms. The Fe3O4@TAPA-BPDA is easy to operate and highly regenerable. A simple magnetic solid-phase extraction (MSPE) method based on the Fe3O4@TAPA-BPDA was then developed for the rapid extraction of five PUHs in real samples, coupled with high-performance liquid chromatography (HPLC) determination. The analytical method developed has a linear range of 0.5-50 ng/mL, and the limit of detection (LOD) ranges from 0.06 to 0.10 ng/mL. The method exhibits good accuracy with recoveries ranged from 74.5 % to 111.4 %. The analytical method was successfully applied to the highly sensitive detection of PUHs in environmental water samples, which highlighting the potential application of the Fe3O4@TAPA-BPDA in the sample pretreatment.

Simultaneous enrichment and sequential elution of cis-diol containing molecules and deoxyribonucleotides with bifunctional boronate and titanium (â £) ion modified-magnetic nanoparticles prior to quantitation by high performance liquid chromatography.

Some diseases can cause abnormal concentrations of catecholamines (CAs), nucleosides (NSs) and nucleotides (NTs) in patients. Previous studies normally focused on the detection of the three types of substances separately. In this work, a bifunctional boronate and titanium (Ⅳ) ion affinity magnetic adsorbent with high-capacity was prepared. The adsorbent can simultaneously enrich CAs, NSs and NTs in a single extraction process, and the adsorbed analytes can be sequentially eluted by 1.0% trifluoroacetic acid and 20.0 mmol L-1 Na3PO4. An analytical method of the analytes has been established by coupling the adsorbent with RP-HPLC. The method has low detection limits (0.039-0.708 ng mL-1) and good reproducibility (inter- and intra-day of assay RSDs less than 15.0%). Serum sample from healthy volunteer was successfully quantified for two CAs, four NSs and five NTs. Compared with the reported methods, the proposed method is simpler to operate, consume less samples, and has enough accurate and sensitivity to obtain comprehensive information on the concentrations of analytes in a single extraction process.

Multivariate covalent organic frameworks guided carboxyl functionalized magnetic adsorbent for enrichment of fluoroquinolones in milk prior to high performance liquid chromatographic analysis.

Herein, we prepared a carboxyl functionalized magnetic covalent organic framework (Fe3O4@iCOF-COOH) by combining multivariate synthetic strategy with post-synthetic modification. It was used as an adsorbent for magnetic solid phase extraction (MSPE) of six fluoroquinolones (FQs), and showed good absorption performance at neutral pH. Carboxyl groups are found to be crucial for the adsorption of fluoroquinolones. The adsorption mechanism was primarily attributed to strong hydrogen bonding, π-π interaction as well as potential hydrophobic effect. The optimal extraction conditions are sample pH at 6.0, adsorbent dosage of 3 mg, eluent of 1.0 mL methanol solution containing 7.5% ammonia, and extraction/desorption time of 30 min. Under the optimized conditions, the Fe3O4@iCOF-COOH was used as an adsorbent for MSPE of FQs in milk, an analytical method was established by combining with high-performance liquid chromatography-ultraviolet detection (HPLC-UV). The limits of detection (LODs) and limit of quantification (LOQs) were 1.24-4.58 ng⋅mL-1 and 4.12-15.3 ng⋅mL-1, respectively. The recoveries of target FQs in spiked milk were 68.4-105%. This work provides a new way to prepare covalent organic framework based adsorbents for solid phase extraction, and can be readily extended to other type of adsorbents.

Preparation of restricted-access boronate affinity adsorbent with excellent anti-protein adsorption property for directly extracting small cis-diol molecules from biological matrices.

Boronate affinity adsorbents are of great promise in the enrichment of small cis-diol-containing molecules (cis-diols) from biological matrices. This work develops a restricted-access boronate affinity mesoporous adsorbent, in which boronate sites are only distributed on the internal surface of mesopores and the external surface is a strongly hydrophilic layer. The adsorbent has high binding capacities (30.3 mg g-1, 22.9 mg g-1 and 14.9 mg g-1 for dopamine, catechol and adenosine, respectively) in spite of removal of the boronate sites on the external surface of adsorbent. The adsorption specific of adsorbent towards cis-diols was assessed by dispersive solid-phase extraction (d-SPE) method, and the results show that the adsorbent can selectively extract small cis-diols in the biosamples while exclude proteins completely. Under the optimal d-SPE, the nucleosides and cis-diol drugs in human serum were successfully analyzed by coupling d-SPE with high-performance liquid chromatography. Where, the detection limits are between 6.1 and 13.4 ng mL-1 for four nucleosides, and 24.9 and 34.3 ng mL-1 for two cis-diol drugs; the relative recoveries of all the analytes vary from 84.1% to 110.1% (RSDs <13.4%, n = 6). The results indicate that the adsorbent can directly treat the real biosamples without the necessary protein precipitation steps in advance, thus simplifying the analysis process.

Hierarchical Mesoporous Metal-Organic Frameworks with Boric Acid Sites on the Inner Surface of Small Mesopores for the Extraction of Nucleotides in Human Plasma Samples.

In this work, a boronate affinity-functionalized hierarchical mesoporous metal-organic framework adsorbent with boronate sites only in the small mesopore has been structured based on UiO-66@Fe3O4. The introduction of large mesopores in the adsorbent can promote the diffusion of small cis-diol-containing compounds (cis-diols) into small mesopore channels, and the removal of the adsorption sites on the external surface of materials and in large mesopores can enhance the size-exclusion effect of the adsorbent. In addition, the adsorbent has faster adsorption kinetics and excellent selectivity to small cis-diols. Finally, a magnetic dispersive solid-phase extraction coupled with high-performance liquid chromatography was established for the enrichment and detection of nucleotides in plasma. Four nucleotides achieve the recoveries from 93.25 to 118.79%, the limits of detection from 0.35 to 1.26 ng·mL-1, and the intra-day and inter-day relative standard deviations of less than 10.2%. In conclusion, this method can be directly used for the detection of small cis-diol targets in complex biological samples without protein precipitation prior to the extraction.

An engineered cascade-sensitized red-emitting upconversion nanoplatform with a tandem hydrophobic hydration-shell and metal-phenolic network decoration for single 808 nm triggered simultaneous tumor PDT and PTT enhanced CDT.

Near-infrared light (NIR) driven lanthanide-doped upconversion nanoparticle (UCNP) based photo-dynamic therapy (PDT) holds a great promise for the non-invasive treatment of deep-seated tumors. However, it has also been highly hindered by the low upconversion luminescence (UCL) efficiency, hypoxia nature of solid tumors, and low therapeutic efficiency using single modality. Herein, we present a novel Nd3+ → Yb3+ → Tm3+ → Er3+ cascade-sensitized red-emitting UCNP with tandem hydrophobic hydration-shell (HHS) and metal-phenolic network (Fe-tannic acid, Fe-TA) decoration (UCNP@HHS@Fe-TA, denoted as UCFS@Fe-TA) for single 808 nm triggered simultaneous tumor PDT and photothermal therapy (PTT) enhanced chemo-dynamic therapy (CDT). The UCNP can supply intense red emission under high tissue penetrating/minimized tissue overheating 808 nm excitation, and their HHS coating with perfluorocarbon/photosensitizer Ce6 co-doping can not only realize UCL-based PDT, but also strengthen PDT of as-formed UCFS via O2-carrying/UCL protection capacity of the HHS. Fe-TA coating can supply 808 nm triggered PTT, and the rise in temperature during PTT leads to enhanced Fenton catalytic activity of Fe-TA and faster ˙OH production rate of CDT to match with the real-timely released 1O2 in PDT. The as-designed UCFS@Fe-TA thus can achieve a single 808 nm triggered simultaneous PDT and PTT enhanced CDT, leading to a PTT-assisted reactive oxygen species storm for efficient tumor suppression. Such a design also renders the nanoplatform lower cell dark toxicity. In addition, the single excitation-triggered multimodal therapy mode might address the excitation wavelength mismatch issue in dual laser-triggered PTT/PDT mode. This study has therefore presented an efficient nanotherapeutic platform enabling synergistic multimodal tumor therapies with high biocompatibility.

Polymer brush grafted immobilized metal ion affinity adsorbent based on polydopamine/polyethyleneimine-coated magnetic graphene oxide for selective enrichment of cytokinins in plants.

An immobilized metal affinity (IMAC) adsorbent was prepared for selective enrichment of adenine type CKs, via grafting polymer chain pendant with iminodiacetic acid (IDA) from polydopamine (PDA)/polyethyleneimine (PEI)-coated magnetic graphene oxide (magGO) via surface-initiated-atom transfer radical polymerization (SI-ATRP). The prepared IMAC sorbent exhibited remarkable adsorption performances and good selectivity for adenine-type CKs and was utilized as a sorbent of magnetic solid-phase extraction (MSPE) for effective enrichment of four adenine-type CKs in bean sprouts. Under the optimized extraction conditions, an analytical method for four adenine type CKs in bean sprouts was established by combining the MSPE combined with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The recoveries of the analytes were between 80.4 ± 1.9% and 114.6 ± 1.5% (n = 3). The limits of detection (LODs) range from 0.63 to 2.30 pg⋅mL-1. The relative standard deviations of intra-day and inter-day were less than 12.6%. The established method was successfully applied to the selective extraction and sensitive detection of trace adenine-type CKs in plant samples.

Effect of stereoconfiguration of aromatic ligands on retention and selectivity of terphenyl isomer-bonded stationary phases.

The structure of ligands has a significant influence on the separation properties of alkyl and aromatic phases in reversed-phase liquid chromatography. Compared with alkyl phases, the effect of stereoconfiguration of aromatic ligands on the retention and selectivity of stationary phases has rarely been addressed. To illustrate the issue, three terphenyl isomer-bonded stationary phases were prepared via the coupling chemistry of isocyanate with terphenyl amine isomers, 3,4-diphenylaniline, 2,4-diphenylaniline and 4-amino-p-terphenyl, respectively. The retention behaviors of stationary phases were assessed in terms of retention strength, selectivity, hydrophobic and π-π interaction by five kinds of solutes. It is found that the selectivity towards the solutes is slightly larger on the branched m-terphenyl-bonded phase (m-π3) than o-terphenyl-bonded phase (o-π3) but is significantly improved on the chain p-terphenyl-bonded phase (p-π3). The results can be interpreted by the ease self-adjustment of the conformation of the chain p-terphenyl ligand and the smaller steric effect of p-π3 towards the insertion of solutes into the ligand brushes. In addition, the p-π3 yields excellent selective separation towards aromatic solutes. These findings are of significance in the design of aromatic stationary phases.

Nitrogen and bromine co-doped carbon dots with red fluorescence for sensing of Ag+ and visual monitoring of glutathione in cells.

Carbon dots (CDs) with red fluorescence emission have excellent advantages in cell imaging. Herein, novel nitrogen and bromine doped CDs (N,Br-CDs) were prepared with 4-bromo-1,2-phenylenediamine as precursor. The N, Br-CDs present the optimal emission wavelength at 582 nm (λex = 510 nm) at pH 7.0 and 648 nm (λex = 580 nm) at pH 3.0 âˆ¼ 5.0, respectively. The fluorescence intensity of N,Br-CDs at 648 nm versus Ag+ concentration shows a good relationship from 0 to 60 µM with the limit of detection (LOD) of 0.14 µM. Furthermore, the fluorescence of N,Br-CDs/Ag+ is efficiently restored via the combination of glutathione (GSH) and Ag+ and linearly changes with GSH concentration from 0 âˆ¼ 6.0 µM with LOD of 49 nM. This method has been successfully employed to monitor intracellular Ag+ and GSH with fluorescence imaging. The results suggest that the N,Br-CDs has application potential in the sensing of Ag+ and visual monitoring of GSH in cells.

Facile Fabrication of Monodisperse Micron-Sized Dual Janus Silica Particles with Asymmetric Morphology and Chemical Environment.

Janus particles are a kind of materials with asymmetric morphology or surface chemical environment. But so far, the preparation of particles with dual asymmetry is still a challenging problem. Hence the cation surfactant hexadecyl trimethyl ammonium bromide and co-surfactant octadecylamine are applied to improve the Pickering emulsion stability, and the micron-sized silica particles are arranged in a single layer at the toluene-water interface through electrostatic interaction. Furthermore, organosilane reagents are added in the preparation process, resulting in the construction of asymmetric hydrophilic or hydrophobic mesoporous precisely onto the micron-sized silica particles surface. The cation surfactant-assisted Pickering emulsion method is simple, effective, and convenience, which can be applied in the synthesis of various dual Janus silica particles for specific applications.

Carbon quantum dots with pH-responsive orange-/red-light emission for fluorescence imaging of intracellular pH.

N-doped carbon quantum dots (N-CDs) with polyaminobenzene hydrazine as precursor were prepared by solvothermal method for the monitoring of pH fluctuation in HeLa cells via fluorescence imaging. The N-CDs show two emission wavelengths at 582 and 640 nm under different pH with two excitation wavelengths. The fluorescence intensity at 640 nm (λex = 520 nm) and the ratio of F582/F640 (λex = 470 nm) linearly increase with pH in the range of 2.4 ~ 3.6 (R2 = 992) and 5.6 ~ 7.6 (R2 = 0.987), respectively. The sensor exhibits high sensitivity and reversibility and anti-interference capability, thus enabling sensing pH change in intracellular environment in real time, as demonstrated by successful monitoring of intracellular pH fluctuation during H2O2 stimulation in HeLa cells.

MOF-coated upconversion nanoconstructs for synergetic photo-chemodynamic/oxygen-elevated photodynamic therapy.

Excessive production of intracellular reactive oxygen species (ROS) can induce apoptosis of cancer cells; however, it is often limited by severe triggering conditions and hypoxic microenvironments of solid tumors. To address these issues, herein, we have designed a MOF-coated upconversion nanoconstruct (UCTSCF, referring to UC@Ce6/TFS@mSiO2@MIL-100(Cu/Fe)) for synergetic photochemodynamic therapy (PCT)/oxygen-elevated photodynamic therapy (PDT). The MOF (MIL-100(Fe)) coating with Cu-doping was designed to catalyze H2O2 overexpression in cancer cells to generate the most cytotoxic ˙OH via chemodynamic therapy (CDT). It is noted that UC, representing 808 nm driven upconversion nanoparticles with high tissue penetration depth/low over-heating effects, was designed to provide intense blue light which can relieve the severe triggering conditions of CDT via PCT. Furthermore, the functional layer of the photosensitizer chlorin e6 (Ce6) and O2-carrying triethoxy(1H,1H,2H,2H-nonafluorohexyl)silane (TFS) co-doped mesoporous silicon (Ce6/TFS@mSiO2) can cause oxygen-elevated 1O2 production upon 671 nm light irradiation. In such a simple ROS generation nanoplatform, we heighten the antitumor effect via oxygen-elevated synergetic tumor PCT/PDT.

In situ reaction-based ratiometric fluorescent assay for alkaline phosphatase activity and bioimaging.

Alkaline phosphatase (ALP) is an important biomarker, it is of great significance to develop a sensitive and efficient analytical method for ALP. In this study, an in situ reaction based ratiometric fluorescence assay for ALP was proposed. l-ascorbic acid-2-phosphate (AA2P) was used as a substrate for ALP, and Cu2+/o-phenylenediamine (OPD) were involved in this system. Cu2+ can oxidize OPD to 2,3-diaminophenazine (OPDox) with an emission centered at 566 nm. The presence of ALP can catalyze the hydrolysis of AA2P to ascorbic acid (AA), which will inhibit the production of OPDox and reduce the corresponding fluorescence intensity, and AA will react with OPD to generate 3-(dihydroxyethyl)furan[3,4-b]quinoxalin-1-one (DFQ) with an emission peak at 447 nm. The fluorescence ratio of F447/F566 has a linear relationship with ALP activity. The proposed method is highly sensitive, finely selective, cost efficiency and easy to operate, it exhibits good linearity in the range of 0.5-22 and 22-40 mU·mL-1, with a detection limit as low as 0.06 mU·mL-1. The excellent applicability of this strategy in human serum samples and MCF-7 cells imaging suggests that this method has promising prospects for biomedical research.

One-step fabrication of nitrogen-rich linear porous organic polymer-based micron-sized sphere for selective enrichment of glycopeptides.

In this work, nitrogen-rich linear porous organic polymers (POPs) were designed to improve glycopeptides enrichment efficiency in HILIC. Two kinds of novel POPs (CH-pPAL and CH-mPAL) were one-step prepared via the Schiff-base condensation reaction and intermolecular hydrogen-bonded using a flexible amine monomer (carbohydrazide, CH) to react with two aldehyde monomers, p-phthalaldehyde (pPAL) and m-phthalaldehyde (mPAL), respectively. The specific surface area of CH-pPAL and CH-mPAL was 238 and 232 m2 g-1, respectively. Based on the hydrogen-bond interaction, several narrow peaks existed in the PXRD patterns. Particularly, micron-sized spherical CH-pPAL possessed a honeycomb-like orderly tunnel structure. The contact angle of CH-pPAL and CH-mPAL severally was 35.4° and 28.8°, respectively, indicating excellent hydrophilicity of as-synthesized POPs. They both demonstrated high sensitivity and high adsorption capacity in glycopeptides enrichment. The 851 and 794 unique glycopeptides derived from 170 and 200 N-glycosylated proteins were identified after the enrichment by CH-pPAL and CH-mPAL within three independent replicates of proteins extracted from human serum, respectively. Therefore, these CH-based hydrophilic POP materials would be applied in the enrichment and identification of low-abundance N-linked glycopeptides from complicated biological samples.

Effect of phenyl numbers in polyphenyl ligand on retention properties of aromatic stationary phases.

Aromatic phase, as one type of reversed-phase stationary phases, shows complementary selectivity to the n-alkyl counterparts especially for certain challenging separation tasks. However, effect of phenyl numbers in aromatic ligands on retention behaviors has rarely been addressed compared with the alkyl stationary phases. To illustrate the issue, a series of polyphenyl stationary phases were facially prepared via the coupling chemistry of isocyanate with amine, including aniline (π1), 4-aminobiphenyl (π2), 4-amino-p-terphenyl (π3) and [1,1':4',1'':4'',1'''-quaterphenyl]-4-amine (π4), respectively. The chromatographic behaviors of the new stationary phases as well as the traditional C18 were systematically compared in terms of retention mode, hydrophobic and aromatic selectivity, shape selectivity and π-π interaction by various analytes, including alkylbenzenes, polycyclic aromatic hydrocarbons congeners and substituted benzenes with electron-withdrawing groups. Due to the homologous structure of four polyphenyl ligands, the hydrophobic selectivity, aromatic selectivity and shape selectivity of stationary phases increase with phenyl numbers in the bonded polyphenyl ligands, whereas the increment becomes insignificant from U-π3 to U-π4. This phenomenon is explained by the insertion degree of analytes in the polyphenyl ligand brushes. Compared with the homemade C18, the polyphenyl phases indicate insignificant changes of shape selectivity with temperature. Notably, the new polyphenyl phases demonstrate the great selective separation towards the electron-deficient compounds through the π-π interaction. These findings make up for the understanding of the retention behavior of aromatic stationary phases.

Preparation of core-shell microporous organic polymer-coated silica microspheres for chromatographic separation and N-glycopeptides enrichment.

Through a "one-pot" strategy, a layer of microporous organic polymer was coated onto the surface of monodisperse amino-functionalized silica microsphere via amino-aldehyde condensation reaction with core-shell structure. The change in chemical structure of material before and after modification was determined by Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy. Due to existence of a large number of amino and aldehyde groups in microporous organic polymer shell, the water contact angle decreased from 56.8° (silica microspheres) to 34.7° (microporous organic polymer-coated silica microspheres). Based on these properties, microporous organic polymer-coated silica microspheres were employed as the stationary phase for capillary liquid chromatography and successfully offered baseline separation of polar small molecules. Additionally, the material could also be served as the sorbent of hydrophilic interaction chromatography to enrich glycopeptides from human serum digest. A total of 470 unique N-glycopeptides and 342 N-glycosylation sites mapped to 112 N-glycosylated proteins were unambiguously identified from 2 µL of human serum, exhibiting a promising application prospect of microporous organic polymer-coated silica microspheres in the pretreatment of proteomics samples.

Removal of adsorption sites on the external surface of mesoporous adsorbent for eliminating the interference of proteins in enrichment of phosphopeptides/nucleotides.

Various mesoporous adsorbents are of great promise for enriching small molecules from biological samples based on the size-exclusion effect. At present, the mesoporous adsorbents have adsorption sites distributed uniformly on the internal and external surfaces of mesopores. However, the adsorption sites on the external surface can adsorb proteins, interfering with the enrichment of small molecules. Herein, a novel immobilized-Ti4+ magnetic mesoporous adsorbent removing the adsorption sites on the external surface was facile prepared via the coupling chemistry of isocyanate with amine and consequent hydrolysis of urea linkage by urease. The adsorbent enables fast and selective enrichment of phosphopeptides and nucleotides from biological samples. In addition, sensitive detection methods for phosphopeptides and nucleotides in human serum are developed by coupling the magnetic solid-phase extraction with matrix-assisted laser desorption/ionization time of flight mass spectrometry and liquid chromatography-mass spectrometer, respectively. Under optimal conditions, response is linear (R2 ≥ 0.9923), limits of detection are low (0.41-9.48 ng mL-1), and reproducibility is acceptable (inter- and intra-day assay RSDs of≤15.0%) for six nucleotides. The developed strategy offers an effective method to eliminate the interference of proteins in the enrichment of small molecules from real biological samples.