Homogeneous immunoassay for soy protein determination in food samples using gold nanoparticles as labels and light scattering detection

A homogeneous aggregation immunoassay involving the use of gold nanoparticles (AuNPs) and light scattering detection is described for soy protein determination in food samples. AuNPs act as enhancers of the precipitate that appears when the antigen-antibody complex is formed. The AuNPs-antibody conjugate has been synthesized by physical adsorption of polyclonal anti-soy protein antibodies onto the surface of commercial AuNPs with a nominal diameter of 20 nm. The direct assay is based on the reaction of the conjugate with soy protein, which reaches the equilibrium in about 10 min, and the measurement of the light scattering intensity at 530 nm, which is proportional to the analyte concentration. The dynamic range of the calibration graph is 0.2-20 μg mL⁻¹ and the detection limit value is 65 ng mL⁻¹. The precision, expressed as relative standard deviation, has been assayed at two different concentrations, 0.2 and 1 μg mL⁻¹, giving values ranging from 4.7 to 5.9%. The interference of other proteins has been assayed. The usefulness of this method has been shown by its application to the analysis of fruit juice and “nonmilk yoghourt” samples. The results obtained with the proposed method are similar to those obtained by using a commercial ELISA kit, but the assay time is significantly shorter and the detection limit was about 10 times lower. A recovery study has been also performed, giving values in the range of 84.0-119.3%.     

Electrode modified with toluidine blue-doped silica nanoparticles, and its use for enhanced amperometric sensing of hemoglobin

Three-dimensionally structured, silica based, organic–inorganic hybrid nanoparticles (NPs) were prepared by a simple and feasible water-in-oil (W/O) microemulsion method and a promising platform for bioelectrochemical analysis was obtained. The commonly used phenathiazine organic compound, toluidine blue (TB) was readily captured in the three-dimensional cage of the inorganic SiO₂ network, which was considered to serve as a protective “shell” toward the embedded TB. A TEM image indicated the size of the thus prepared TB-doped SiO₂ (TB@SiO₂) NPs was 21 ± 3 nm. UV–visible and IR spectroscopy confirmed successful formation of the organic–inorganic composite and possible interaction between TB and SiO₂, which favored enhanced stability of the hybrid. A sensitive amperometric sensor for hemoglobin (Hb) biomolecules based on TB@SiO₂ NPs conjugated with a biopolymer chitosan (CHIT) membrane was then developed. The surface of the silica NPs was highly biocompatible and the TB captured inside maintained its high electron-transfer efficiency. Dye leakage of TB from the TB@SiO₂ hybrid was proved to be minimal, owing to the inorganic SiO₂ network and the force of interaction between TB and SiO₂. The amperometric sensor had a detection limit of 2.5 × 10⁻⁹ mol L⁻¹ (S/N = 3) with a linear range from 5.0 × 10⁻⁹ to 3.0 × 10⁻⁶ mol L⁻¹ for Hb. When it was applied to determine the concentration of a clinical blood sample, satisfactory results were obtained which were in good agreement with those obtained by the standard method.     

Long-wavelength homogeneous enzyme immunoassay for the determination of amikacin in water samples

A simple and rapid homogeneous enzyme immunoassay involving the use of the malic dehydrogenase enzyme and a long-wavelength fluorophor, the oxazine Cresyl Violet, is proposed for the determination of the antibiotic amikacin in water samples. An enzymatic tracer has been synthesized by covalent binding of amikacin to malic dehydrogenase via a carbodiimide derivative. Free tracer catalyses the reaction between Cresyl Violet and malic acid giving rise to a decrease in the fluorescence of the fluorophor. Kinetic curves for this reaction have been monitored at λ_ex 585 and λ_em 624 nm using the stopped-flow mixing technique, being the initial rate measured in only 2-3 s. The dynamic range of the method is 1-15 ng mL⁻¹ and the detection limit is 0.3 ng mL⁻¹, using aqueous standard solutions or water samples. The precision, obtained at 1 and 5 ng mL⁻¹ and expressed as relative standard deviation, was 6.0 and 9.6%, respectively. The method has been applied to the analysis of drinking, river and wastewater samples. The sample pre-treatment involved a solid-phase extraction step for the clean-up of the samples. A recovery study was carried out to validate the method, being the values obtained in the range 80-114%, with a mean value of 96.7%.     

A study of using luminophore-doped silica nanoparticles as fluorescent probe in protein microarray assay

A convenient method for the synthesis of tris(2,2′-bipyridyl) dichlororuthenium(II) hexahydrate-doped amino-modified double-layer silica nanoparticles is presented in this paper. The synthesized nanoparticles are uniform and photostable, and can be well dispersed in a water solution. Proteins could be directly immobilized onto these nanoparticles by a simple coupling process without losing their biological activities. These nanoparticles were further used as fluorescent probes in protein microarray assay for the quantitative detection of protein. The results obtained by these nanoparticles, with the detection limit of as low as 3.5 μg/mL, were much better than those involving the use of conventional FITC probe. Translated from Chinese Journal of Analytical Chemistry, 2006, 34(9): 1227–1230 (in Chinese)     

Copper nanoclusters conjugated silica nanoparticles modified on carbon paste as an electrochemical sensor for the determination of dopamine

An electrochemical sensor based on modification of carbon paste electrode by glutathione‐capped copper nanoclusters silica nanoparticles (CuNCs/SiO₂NPs) composite for determination of dopamine in the presence of ascorbic acid was presented. Transmission electron microscopy, scanning electron microscopy, energy dispersive X‐Ray analysis, X‐ray photoelectron spectroscopy, Fourier‐transform infrared spectroscopy, X‐ray diffraction and electrochemical impedance spectroscopy were used for characterization of the developed electrode. The electrochemical behavior of dopamine on CuNCs/SiO₂NPs/carbon paste electrode was investigated by cyclic voltammetry and differential pulse voltammetry. Dopamine was determined in the range of 10.0 – 900.0 μM, and the limit of detection was obtained as 0.43 μM. The electrochemical behaviors of the coexisting electroactive species, which often cause interference with the determination of dopamine, were investigated. The results show that the developed electrode does not show any interference with respect to coexisting species, even in the presence of ascorbic acid. The developed electrochemical sensor was further employed for the determination of dopamine in human blood plasma, with a good recovery.     

Lipid,protein and poly(NIPAM) coated mesoporous silica nanoparticles for biomedical applications

In the past decade, mesoporous silica nanoparticles (MSNs) as nanocarriers have showed much potential in advanced nanomaterials due to their large surface area and pore volume. Especially, more and more MSNs based nanodevices have been designed as efficient drug delivery systems (DDSs) or biosensors. In this paper, lipid, protein and poly(NIPAM) coated MSNs are reviewed from the preparation, properties and their potential application. We also introduce the preparative methods including physical adsorption, covalent binding and self-assembly on the MSNs' surfaces. Furthermore, the interaction between the aimed cells and these molecular modified MSNs is discussed. We also demonstrate their typical applications, such as photodynamic therapy, bioimaging, controlled release and selective recognition in biomedical field.     

Palladium nanoparticles immobilized on Schiff base‐functionalized mesoporous silica as a highly efficient and magnetically recoverable nanocatalyst for Heck coupling reaction

A new magnetically recoverable nanocatalyst was prepared by functionalization of mesoporous silica (SBA‐15) with a Schiff base ligand, and then immobilization of palladium nanoparticles on it using a simple procedure. This heterogeneous catalyst was fully characterized using appropriate analyses and its catalytic efficiency was investigated in Heck reaction using iodo‐, bromo‐ and chlorobenzene derivatives and styrene, with the aim of synthesizing stilbene derivatives, a class of compounds with a variety of pharmacological properties. Some of the characteristics of this nanocatalyst include good dispersion of palladium nanoparticles on the SBA‐15 support, easy separation, catalyses the production of stilbene derivatives in a short time with excellent yields even for bromo‐ and chlorobenzene, and preservation of its catalytic activity after eight reaction cycles.     

Development of superparamagnetic high-magnetization C18-functionalized magnetic silica nanoparticles as sorbents for enrichment and determination of methylprednisolone in rat plasma by high performance liquid chromatography

In this study, a novel extraction and enrichment technique based on superparamagnetic high-magnetization C₁₈-functionalized magnetic silica nanoparticles (C₁₈-MNPs) as sorbents was successfully developed for the determination of methylprednisolone (MP) in rat plasma by high performance liquid chromatography (HPLC). The synthesized silica-coated magnetite modified with chlorodimethyl-n-octadecylsilane was about 320 nm in diameter with strong magnetism and high surface area. It provided an efficient way for extraction and concentration of MP in the samples through hydrophobic interaction by the interior C₁₈ groups. Moreover, MP adsorbed with C₁₈-MNPs could be simply and rapidly isolated through placing a strong magnet on the bottom of container, and then easily eluted from C₁₈-MNPs by n-hexane solution. Extraction conditions such as amounts of C₁₈-MNPs added, adsorption time and desorption solvent, were investigated. Method validations including linear range, detection limit, precision, and recovery were also studied. The results showed that the proposed method based on C₁₈-MNPs was a simple, accurate and high efficient approach for the analysis of MP in the complex plasma samples.     

Imprinted silica nanoparticles coated with N‐propylsilylmorpholine‐4‐carboxamide for the determination of m‐cresol in synthetic and real samples

m‐Cresol‐imprinted silica nanoparticles coated with N‐propylsilylmorpholine‐4‐carboxamide have been developed that contain specific pockets for the selective uptake of m‐cresol. Silica nanoparticles were synthesized by a sol–gel process followed by functionalization of their surface with N‐propylsilylmorpholine‐4‐carboxamide. The formation of m‐cresol‐imprinted silica nanoparticles was confirmed by UV‐Vis spectrophotometry, infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy and transmission electron microscopy. Electron microscopic studies revealed the formation of monodispersed imprinted silica nanoparticles with spherical shape and an average size of 83 nm. The developed nanoparticles were filled in a syringe and used for the extraction of m‐cresol from aqueous samples followed by quantification using high‐performance liquid chromatography with diode array detection. Various adsorption experiments showed that developed m‐cresol‐imprinted silica nanoparticles exhibited a high adsorption capacity and selectivity and offered a fast kinetics for rebinding m‐cresol. The chromatographic quantification was achieved using mobile phase consisting of acetonitrile/water (70:30 v/v) at an isocratic flow rate of 1.0 mL/min using a reversed‐phase C₁₈ column and detection at λ_max = 275 nm. The limits of detection and quantification were 1.86 and 22.32 ng/mL, respectively, for the developed method. The percent recoveries ranged from 96.66–103.33% in the spiked samples. This combination of this nanotechnique with molecular imprinting was proved as a reliable, sensitive and selective method for determining the target from synthetic and real samples.