Zirconia layer coated mesoporous silica microspheres used for highly specific phosphopeptide enrichment

Zirconia layer coated mesoporous silica microspheres with mesostructured cellular foams (MCFs) were prepared by NH₃/water vapor-induced internal hydrolysis method. Zirconia layer coated MCF microspheres were characterized by SEM, XRD, N₂ sorption, UV, and chromatographic analysis, and explored for enrichment of phosphopeptides. ZrO₂/MCF microspheres in solid-phase extraction (SPE) mode demonstrated much higher selectivity and higher efficiency towards phosphopeptide enrichment than bulk ZrO₂ particles. In particular, the selectivities of ZrO₂/MCF microspheres towards multi-phosphopeptides are even higher than that of the widely used commercial TiO₂ microparticles. The ZrO₂/MCF microspheres were also applied to enrich endogenous phosphopeptides from human serum, and twelve endogenous phosphorylated peptides could be specifically enriched.     

Sequestration of CO2 using Cu nanoparticles supported on spherical and rod-shape mesoporous silica

The CO₂ sequestration is one of the most promising solutions to tackle global warming. In this study, spherical mesoporous silica particles (MPS-S) and rod-shaped mesoporous silica particles (MPS-R) loaded with Cu nanoparticles were selectively prepared and employed for CO₂ adsorption. For the first time uniform Cu nanoparticles were incorporated into the rod-shaped mesoporous silica particles by post-synthesis modification using both N-[3-(trimethoxysilyl)propyl]ethylenediamine (PEDA) and ethylenediamine (EDA) as coupling agents. The physiochemical properties of the mesoporous and copper grifted silica composites were investigated by CHN elemental analysis, FTIR spectroscopy, thermogravimetric analysis, X-ray diffraction, energy dispersive X-ray spectroscopy (EDX), surface area analysis, scanning, transmission electron microscopy and gas analysis system (GSD 320, TERMO). The mesoporous silica shows highly ordered mesoporous structures, with the rod-shaped particles having a higher surface area than the spherical ones. Copper nanoparticles with an average diameter of 6.0 nm were uniformly incorporated into the MPS-S and MPS-R. Moreover, Cu-loaded mesoporous silica exhibits up to 40% higher CO₂ adsorption capacity than the bare MPS. The MPS-R modified with Cu nanoparticles showed a maximum CO₂ adsorption capacity of 0.62 mmol/g and the humidity showed a slight negative effect on CO₂ uptake process. The enhancement of CO₂ adsorption onto transition metal/mesoporous substrates provides basis for imminent CO₂ sequestration.     

Isolation of phosphopeptides using solid phase enrichment

Protein phosphorylations are post-translational protein modifications that are crucial for intercellular signal transduction and the regulation of many cellular events. We describe herein a novel technology utilizing α-diazo functionalized solid phase resins to isolate phosphorylated peptides from non-phosphorylated substrates.     

An integrated procedure of selective injection, sample stacking and fractionation of phosphopeptides for MALDI MS analysis

Protein phosphorylation is one of the most important post-translational modifications (PTM), however, the detection of phosphorylation in proteins using mass spectrometry (MS) remains challenging. This is because many phosphorylated proteins are only present in low abundance, and the ionization of the phosphorylated components in MS is very inefficient compared to the non-phosphorylated counterparts. Recently, we have reported a selective injection technique that can separate phosphopeptides from non-phosphorylated peptides due to the differences in their isoelectric points (pI) [1]. Phosphorylated peptides from α-casein were clearly observed at low femtomole level using MALDI MS. In this work, further developments on selective injection of phosphopeptides are presented to enhance its capability in handling higher sample complexity. The approach is to integrate selective injection with a sample stacking technique used in capillary electrophoresis to enrich the sample concentration, followed by electrophoresis to fractionate the components in preparation for MALDI MS analysis. The effectiveness of the selective injection and stacking was evaluated quantitatively using a synthetic phosphopeptide as sample, with an enrichment factor of up to 600 being recorded. Next, a tryptic digest of α-casein was used to evaluate the separation and fractionation of peptides for MALDI MS analysis. The elution order of phosphopeptides essentially followed the order of decreasing number of phosphates on the peptides. Finally, to illustrate the applicability, the integrated procedure was applied to evaluate the phosphorylation of a highly phosphorylated protein, osteopontin. Up to 41 phosphopeptides were observed, which allowed us to examine the phosphorylation of all 29 possible sites previously reported [2]. A high level of heterogeneity in the phosphorylation of OPN was evident by the multiple-forms of variable phosphorylation detected for a large number of peptides.     

Thermooxidative stability of polymethyl methacrylate containing nanoparticles of silica/titania and silica/zirconia

The influence of silica/titania and silica/zirconia nanoparticles on thermooxidative degradation of PMMA was studied by non-isothermal thermogravimetry. Kinetic parameters describing the length of the oxidation induction periods were obtained from the treatment of the dependence of oxidation onset temperature on heating rate. Using these parameters, the protection factors of nanoparticles have been calculated. It was found that SiO₂/TiO₂ nanoparticles increase the thermooxidation stability of PMMA where the stabilizing effect, expressed as the protection factor, depends on temperature only slightly. The stabilizing effect of SiO₂/ZrO₂ is much stronger than that of SiO₂/TiO₂ and decreases with increasing temperature.     

Label-free detection of adenosine based on fluorescence resonance energy transfer between fluorescent silica nanoparticles and unmodified gold nanoparticles

A sensitive and convenient strategy was developed for label-free assay of adenosine. The strategy adapted the fluorescence resonance energy transfer property between Rhodamine B doped fluorescent silica nanoparticles (SiNPs) and gold nanoparticles (AuNPs) to generate signal. The different affinities of AuNPs toward the unfolded and folded aptamers were employed for the signal transfer in the system. In the presence of adenosine, the split aptamer fragments react with adenosine to form a structured complex. The folded aptamer cannot be adsorbed on the surface of AuNPs, which induces the aggregation of AuNPs under high ionic concentration conditions, and the aggregation of AuNPs leads to the decrease of the quenching ability. Therefore, the fluorescence intensity of Rhodamine B doped fluorescent SiNPs increased along with the concentration of adenosine. Because of the highly specific recognition ability of the aptamer toward adenosine and the strong quenching ability of AuNPs, the proposed strategy demonstrated good selectivity and high sensitivity for the detection of adenosine. Under the optimum conditions in the experiments, a linear range from 98 nM to 100 μM was obtained with a detection limit of 45 nM. As this strategy is convenient, practical and sensitive, it will provide a promising potential for label-free aptamer-based protein detection.     

Core-shell effects of functionalized oxide nanoparticles inside long-range meso-ordered spray-dried silica spheres

Core-shell and homogeneous distributions of functionalized cerium oxide nanoparticles within spray-dried mesostructured silica spheres are achieved by modification of synthesis parameters such as the templating agent and nanoparticle capping functions. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.

Application of liquid phase deposited titania nanoparticles on silica spheres to phosphopeptide enrichment and high performance liquid chromatography packings

A novel core-shell composite (SiO(2)-nLPD), consisting of micrometer-sized silica spheres as a core and nanometer titania particles as a surface coating, was prepared by liquid phase deposition (LPD). Here, we show the resulting core-shell composite to have better efficient and selective enrichment for mono- and multi-phosphopeptides than commercially available TiO(2) spheres without any enhancer. The material exhibited favorable characteristics for HPLC, which include narrow pore size distribution, high surface area and pore volume. We also show that the core-shell composite can efficiently separate adenosine phosphate compounds due to the Lewis acid-base interaction between titania and phosphate group when used as HPLC packings. After coating the silica sphere with titania by LPD, the silanol of silica spheres will be shielded and that the stationary phase, C(18) bonded SiO(2)-3LPD, could be used under extreme pH condition.     

Development of an on-column enrichment technique based on C18-functionalized magnetic silica nanoparticles for the determination of lidocaine in rat plasma by high performance liquid chromatography

In this study, a novel on-column enrichment technique filled with C(18)-functionalized magnetic silica nanoparticles was successfully developed for the determination of lidocaine in rat plasma by high performance liquid chromatography (HPLC). The synthesized Fe(3)O(4)@SiO(2)-C(18) nanoparticles were locally packed into the capillary by the application of magnets. Lidocaine in the sample solutions pumped into the capillary tube could be easily adsorbed by Fe(3)O(4)@SiO(2)-C(18) through hydrophobic interaction by the interior C(18) groups, and eluted by acetonitrile solution. Different extraction conditions were investigated. Method validations including linear range, quantification limit, detection limit, precision, accuracy and recovery were also studied. The results showed that the proposed method based on on-column enrichment by Fe(3)O(4)@SiO(2)-C(18) was a novel, little solvent and efficient approach for the determination of lidocaine in the complex plasma samples.     

Functionalized diamond nanopowder for phosphopeptides enrichment from complex biological fluids

Diamond is known for its high affinity and biocompatibility towards biomolecules and is used exclusively in separation sciences and life science research. In present study, diamond nanopowder is derivatized as Immobilized Metal Ion Affinity Chromatographic (IMAC) material for the phosphopeptides enrichment and as Reversed Phase (C-18) media for the desalting of complex mixtures and human serum profiling through MALDI-TOF-MS. Functionalized diamond nanopowder is characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) spectroscopy. Diamond-IMAC is applied to the standard protein (β-casein), spiked human serum, egg yolk and non-fat milk for the phosphopeptides enrichment. Results show the selectivity of synthesized IMAC-diamond immobilized with Fe³⁺ and La³⁺ ions. To comprehend the elaborated use, diamond-IMAC is also applied to the serum samples from gall bladder carcinoma for the potential biomarkers. Database search is carried out by the Mascot program (www.matrixscience.com) for the assignment of phosphorylation sites. Diamond nanopowder is thus a separation media with multifunctional use and can be applied to cancer protein profiling for the diagnosis and biomarker identification.     

Preparation of well-defined polystyrene/silica hybrid nanoparticles by ATRP

Immobilization of the atom transfer radical polymerization (ATRP) macroinitiators at the silica nanoparticle surfaces was achieved through surface modification with excess toluene-2,4-diisocynate (TDI), after which the residual isocyanate groups were converted into ATRP macroinitiators. Structurally well-defined polystyrene chains were grown from the nanoparticle surfaces to yield individual particles composed of a silica core and a well-defined, densely grafted outer polystyrene by ATRP, which was initiated by the as-synthesized silica-based macroinitiator. FTIR, NMR and gel permeation chro-matography (GPC) were used to characterize the polystyrene/silica hybrid particles.     

Effect of silica nanoparticles with different sizes on the catalytic activity of glucose oxidase

In this work we present a strategy for the covalent immobilization of periodate oxidized glucose oxidase () to aminated silica nanoparticles (ASNPs) modified on gold electrodes. Silica nanoparticles greatly enhanced the catalytic ability of GOx toward the oxidation of glucose and improved the electron transfer between the GOx and the electrode surface. ASNPs of varying size—that is 100, 80, 60, and 30 nm—were prepared, and they were used to fabricate biosensors. Electrochemical impedance spectroscopy (EIS) of ferrocyanide followed the assembly process and verified the successful immobilization of on ASNPs modified on gold electrodes. From the analysis of catalytic signals of biosensors using different sizes of ASNPs under the same conditions, the surface concentration of electrically wired enzyme (Γ _ET) was estimated and was found to increase with decreasing ASNPs size. Therefore, the sensitivity of biosensors using smaller ASNPs was higher than that using larger particles. Specifically, we utilized the ASNPs with optimal size (30 nm) to fabricate the glucose biosensor. The resulting electrodes showed a wide linear response to glucose at least to 6 mM and reached 95% of the steady-state current in less than 4 s with a sensitivity of 5.02 μA mM⁻¹ cm⁻² and a detection limit of 0.01 mM. The biosensor also showed excellent stability and good reproducibility. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Heterogeneous immunoassay for soy protein determination using nile blue-doped silica nanoparticles as labels and front-surface long-wavelength fluorimetry

A long-wavelength fluoroimmunoassay for the determination of soy protein is reported for the first time using a conjugate composed of anti-soy protein antibodies bound to nile blue-doped silica nanoparticles (NPs). These NPs have been synthesized by a reverse-micelle microemulsion method and functionalized by using 3-(aminopropyl)triethoxysilane (APS) and 3-(trihydroxysilyl)propyl methylphosphonate (THPMP) to avoid NP aggregation. The tracer has been obtained by linking the functionalized NPs with anti-soy protein antibodies previously oxidised with sodium periodate. The immunoassay has been developed in 96-well microplates using a heterogeneous competitive format with antibody capture. Soy proteins are immobilised onto the wells and bovine serum albumin is added to block the surface, thus minimising non-specific binding. After washing, the microplates can be stored ready to use. At the analysis time, soy protein standards or sample and tracer are added and incubated and, after the corresponding washing and drying steps, the fluorescence is measured onto the solid surface at λ_ex 620 and λ_em 680 nm. The method features a dynamic range of 0.1–10 mg L⁻¹ and a detection limit of 0.05 mg L⁻¹. The precision of the method has been assayed at 0.5 and 5 mg L⁻¹ protein concentrations, obtaining the values of relative standard deviation of 9.6% and 6.1%, respectively. This new immunoassay has been applied to the analysis of food containing soy protein and the results obtained have been compared to those provided by a commercial ELISA kit with no statistically differing results. Also, a recovery study has been performed, providing percentages in the range of 81.5–111.0%.     

In situ radical transfer addition polymerization of styrene from silica nanoparticles

The in situ radical transfer addition polymerization of styrene from silica nanoparticles was carried out by the free radical polymerized of styrene in the presence of mercaptopropyl-modified silica nanoparticles as chain-transfer agent. The effects of the amount of the initiator, polymerizing temperature and polymerizing time on the convention of styrene (C) and the percentage of grafting were investigated. Results of elemental analysis, IR, X-ray photoelectron spectrometer and transmission electron microscope demonstrated that the desired polymer chains have been covalently bonded to the surface of the silica nanoparticles. A C of 42.56% and a PG of 38.10% could be achieved with the optimal condition. The polystyrene grafted silica nanoparticles could be separated and used as nanofiller for polymers.     

Green preparation of hollow mesoporous silica nanosphere inside-loaded gold nanoparticles and the catalytic activity

In this work, an active nano-catalyst with gold nanoparticles loaded in hollow mesoporous silica nanospheres (HMSNs/Au) was prepared by a one-pot sol-gel method, in which gold ions were loaded in hollow mesoporous silica spheres followed by sodium alginate reduction. The characterization of the HMSNs/Au were determined by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), N₂ adsorption–desorption isotherms (BET). The high catalytic activity of HMSNs/Au, denoted as apparent turn-over frequency (TOF), was detected by UV-Vis spectrophotometer for the catalytic reduction of 4-nitrophenol (74.5 h^?1) and 2-nitrophenol (108.7 h^?1) in the presence of sodium borohydride solution due to the small gold nanoparticles size and overall exposure of active sites. It is expected that this ecofriendly approach to prepare inorganic composited nanoparticles as high active catalysts based on hollow mesoporous materials was a promising platform for loading noble metal nanoparticles.     

Solid state synthesis of water-dispersible silicon nanoparticles from silica nanoparticles

A solid state synthesis for obtaining nanocrystalline silicon was performed by high temperature reduction of commercial amorphous nanosilica with magnesium powder. The obtained silicon powder contains crystalline silicon phase with lattice spacings characteristic of diamond cubic structure (according to high resolution TEM), and an amorphous phase. In ²⁹Si CP MAS NMR a broad multicomponent peak corresponding to silicon is located at −61.28 to −69.45 ppm, i.e. between the peaks characteristic of amorphous and crystalline Si. The powder has displayed red luminescence while excited under UV illumination, due to quantum confinement within the nanocrystals. The silicon nanopowder was successfully dispersed in water containing poly(vinyl alcohol) as a stabilizing agent. The obtained dispersion was also characterized by red photoluminescence with a band maximum at 710 nm, thus enabling future functional coating applications.     

Palladium nanoparticles supported on poly (N-vinylpyrrolidone)-grafted silica as new recyclable catalyst for Heck cross-coupling reactions

Novel catalytic system based on palladium nanoparticles supported on poly (N-vinylpyrrolidone) (PVP) grafted silica was prepared. Aminopropylsilica was reacted with acryloyl chloride to form acrylamidopropylsilica, and onto this functionalized silica vinylpyrrolidone monomer was polymerized by free-radical polymerization. The complexation of PVP-grafted silica with PdCl₂ was carried out to obtain the heterogeneous catalytic system. X-ray diffraction (XRD) technique and transmission electron microscopy (TEM) image showed that palladium dispersed through the support in nanometer size. This catalytic system exhibited excellent activity in cross-coupling reactions of aryl iodides, bromides and also chlorides with olefinic compounds in Heck-Mizoraki reactions in short reaction time and high yields. Elemental analysis of Pd by inductively coupled plasma (ICP) technique and hot filtration test showed low leaching of the metal into solution from the supported catalyst. The catalyst can be reused several times in repeating Heck reaction cycles without considerable loss in its activity.     

The effect of silica nanoparticles on the stability of aqueous foams

An experimental study was performed on aqueous foams stabilized by a mixture of hexadecyltrimethylammonium bromide (HTAB) and negatively-charged silica nanoparticles. The effects of the nanoparticles on the stability of foams at different HTAB concentrations were investigated. The foams were characterized by measuring their foamability and stability. Rheological behavior of the foams was also studied. Furthermore, rheology of the air–water interfaces was studied in the linear and nonlinear deformation ranges. The thickness of the monolayer at the interface was measured. The actual size of the silica nanoparticles at the air–water interface was measured by transmission electron microscopy. Based on these measurements, the interaction between the monolayers across the foam film containing HTAB and nanoparticles was investigated. Smaller silica nanoparticles (i.e. diameter less than 10?nm) adsorbed at the air–water interface whereas the larger particles remained in the sub-phase or in the bulk liquid phase. It was found that these nanoparticles strongly influenced the foaming behavior at the low HTAB concentrations (i.e. below the CMC). A Langmuir-type monolayer was formed. The presence of the nanoparticles at the air–water interface provided mechanical strength to the foam films and prevented their rupture. This hindered coalescence of the bubbles, which resulted in a stable foam.     

Characterization of gold nanoparticles electrochemically deposited on amine-functioned mesoporous silica films and electrocatalytic oxidation of glucose

This study reports the preparation and characterization of gold nanoparticles deposited on amine-functioned hexagonal mesoporous silica (NH₂–HSM) films and the electrocatalytic oxidation of glucose. Gold nanoparticles are fabricated by electrochemically reducing chloroauric acid on the surface of NH₂–HSM film, using potential step technology. The gold nanoparticles deposited have an average diameter of 80 nm and show high electroactivity. Prussian blue film can form easily on them while cycling the potential between −0.2 and 0.6 V (vs saturated calomel electrode) in single ferricyanide solution. The gold nanoparticles loading NH₂–HSM-film-coated glassy carbon electrode (Au–NH₂–HSM/GCE) shows strong catalysis to the oxidation of glucose, and according to the cathodic oxidation peak at about 0.16 V, the catalytic current is about 2.5 μA mM⁻¹. Under optimized conditions, the peak current of the cathodic oxidation peak is linear to the concentration of glucose in the range of 0.2 to 70 mM. The detection limit is estimated to be 0.1 mM. In addition, some electrochemical parameters about glucose oxidation are estimated.     

Preparation of ultrathin coating layers using surface modified silica nanoparticles

Silica nanoparticles are used in various applications including catalysts, paints and coatings. To reach an optimal performance via stability and functionality, in most cases, the surface properties of the particles are altered using complex procedures. Here we describe a simple method for surface modification of silica nanoparticles (SNP) using sequential adsorption of oppositely charged components. First, the SNPs were made cationic by adsorption of a cationic polyelectrolyte. Poly(allylamine hydrochloride) (PAH) and polyethyleneimine (PEI) were chosen as polycations to investigate the difference between a linear and a branched polyelectrolyte. Next, the dispersion of cationic SNPs was combined with an anionic alkyl ketene dimer (AKD) emulsion. Using this approach cationic, hydrophobic silica particle dispersions were produced. Dynamic light scattering, contact angle measurements and atomic force microscopy (AFM) were used for analyzing the particle and coating layer properties. The chosen polyelectrolyte affected the structure of the dispersion. The layer build-up was studied in detail using a quartz crystal microbalance with dissipation monitoring (QCM-D). The adsorption and layer properties of the cationic polyelectrolytes adsorbed on silica as well as the affinity of AKD to this layer were explored. The application possibilities of the modified particle dispersions were demonstrated by preparing paper and silica surfaces with tailored properties, such as elevated surface hydrophobicity, using an ultrathin coating layer.