Highly sensitive determination of trace potassium ion in serum using the resonance light scattering technique with sodium tetraphenylboron

A resonance light scattering technique has been developed in order to determine potassium ion in serum. Potassium ion was found to bind the tetraphenylboronate anion [(C₆H₅)₄B]⁻ in acetate buffer (pH 8.0) in the presence of sodium dodecyl benzene sulfonate as a stabilizer, forming the B(C₆H₅)₄-K aggregate which produces intense resonance scattering light. Effects of factors such as acidity, ionic strength and interferents on the RLS of B(C₆H₅)₄-K were investigated. The solution pH close to neutral facilitates the production of RLS, and few biologically relevant species interfere in the determination of potassium ion. The resonance scattering light intensity at the maximum peak of 567 nm was linear to the concentration of potassium ion in the range of 0.2–2.0 μg mL⁻¹ with a detection limit of 20.0 ng mL⁻¹. The method was applied to determine trace amounts of potassium ion in serum and showed high sensitivity and accuracy compared with the clinically used ion-selective electrode method.     

Interactions of norfloxacin with DNA and determination of DNA at nanogram levels based on the measurement of enhanced resonance light scattering

A novel assay of DNA at nanogram levels is presented that is based on the measurement of resonance light scattering (RLS) signals in the presence of norfloxacin. The characteristics of RLS spectra, the effective factors and the optimum conditions of the reaction have been investigated. In Britton-Robinson buffer (pH 5.87), norfloxacin has a maximum peak at 405.5 nm and the RLS intensity is greatly enhanced by trace amounts of DNA due to the interaction between norfloxacin and DNA. Mechanistic studies show that the binding of norfloxacin to DNA forms large particles, which result in the significant enhancement of RLS intensity. The enhanced intensity of RLS is proportional to the concentration of DNA in the range from 0.01–2.0 μg mL⁻¹ for yeast DNA, and from 0.02 to 2.3 μg mL⁻¹ for calf thymus DNA. The determination limit (3σ) is 0.7 ng mL⁻¹ for yeast DNA and 1.2 ng mL⁻¹ for calf thymus DNA, respectively. Synthetic samples were determined satisfactorily.     

Application of Functionalized Ag Nanoparticles for the Determination of Proteins at Nanogram Levels Using the Resonance Light Scattering Method

A novel method for the determination of proteins in aqueous solutions has been developed based on the enhancement of resonance light scattering (RLS) of Ag nanoparticles in the presence of proteins. Factors including acidity of the media, concentration of Ag hydrosol, reaction time, temperature, and interference of non-protein substances were investigated. Under the optimal conditions, with the enhanced RLS signals at 452nm, the linear ranges of calibration curves were 0–0.8µgmL^–1 for bovine serum albumin (BSA), 0–1.2µgmL^–1 for human serum albumin (HSA), and 0–2.5µgmL^–1 for human -IgG (-IgG), respectively. The detection limits were 1.3ngmL^–1 for BSA, 10ngmL^–1 for HAS, and 5.7ngmL^–1 for -IgG.This method has been applied to the analysis of synthetic samples and real human serum samples, and the results were in good agreement with those reported by the hospital, indicating that the method presented here is not only sensitive and simple, but also reliable and suitable for practical applications.     

Determination of Nucleic Acid Using the Resonance Light Scattering Technique with the Mixed Complex La(phen)(o-phthalic acid)

A mixed complex of lanthanum with o-phenanthroline (phen) and o-phthalic acid (C₈H₆O₄) was synthesized. The resonance light scattering (RLS) and UV characteristics of interaction between La(phen)(C₈H₆O₄) and ctDNA were studied. Based on the enhancement of RLS intensity of La(phen) (C₈H₆O₄) by DNA at 400nm in the pH range of 6.5 to 7.5, a new method for determination of nucleic acids has been developed. The calibration curve was linear in the range of 0.023 to 1.87µgmL^–1. The detection limit (3) was 0.023µgmL^–1. Interference by coexisting substances in the determination of DNA was also examined. Three synthetic samples were determined with satisfaction.     

Determination of Yttrium by Resonance Light Scattering Technique

 Yttrium was determined by means of the resonance light scattering (RLS) technique. The characteristics of resonance light scattering spectra of yttrium-1, 6-bi(1′-phenyl-3′-methyl-5′-pyrazolone-4′-) hexanedione (BPMPHD), the effective factors and optimum conditions were studied. In the pH range of 5.0–6.1, yttrium-BPMPHD complex produces three characteristic peaks of RLS at 365, 402 and 467 nm. The enhanced intensity of RLS is proportional to the concentration of yttrium in the range of 1.0×10⁻⁸ to 1.0×10⁻⁵ mol · L⁻¹. The limit of detection is 5.9×10⁻⁹ mol · L⁻¹. The method has been used for the determination of Y³⁺ in mixed rare earths. Correspondence: Key Lab for Colloid and Interface Chemistry of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, People’s Republic of China. e-mail: wux@sdu.edu.cn Received July 3, 2002; accepted October 20, 2002     

Determination of Proteins at Nanogram Levels Using the Resonance Light Scattering Technique with a Novel PVAK Nanoparticle

A novel functionalized polyvinyl alcohol keto-derivative nanoparticle (PVAK) has been prepared in a one-step method using oxidation and degradation under ultrasonic irradiation. The nanoparticle is water-soluble, chemically stable, non-toxic and biocompatible. The surface of the nanoparticle is covered with abundant hydroxyl, carbonyl and carboxyl. At pH 3.0, the interactions of PVAK with different proteins can result in obviously enhanced RLS signals at 380nm. Under optimal conditions, the calibration graphs are linear over the range of 0.024.0µgmL^–1 for human serum albumin (HSA), 0.023.5µgmL^–1 for bovine serum albumin (BSA), and 0.053.5µgmL^–1 for human -globulin (-G), respectively. Detection limits were 6.4ngmL^–1 for HSA, 9.2ngmL^–1 for BSA, and 12.5ngmL^–1 for -G, respectively. The method was employed for the determination of total proteins in human serum with satisfactory results.     

SPR imaging as a tool for detecting mucin – anti-mucin interaction. Outline of the development of a sensor for near-patient testing for mucin

Sensors able to provide ‘yes’/‘no’ answers have become of interest in recent years, especially in the fields of environmental research and healthcare. We describe a procedure based on surface plasmon resonance imaging (SPRI) to investigate the interaction between mucin and anti-mucin antibody, and we outline the development of an alarm sensor for the protein mucin, whose high concentration in saliva, blood or tissue is related to illnesses such as gingivitis, peridontitis or even cancer. Anti-mucin gastric antibodies are immobilised onto a gold surface. The immobilisation is evaluated for neat gold chips and for polymer-modified gold surfaces. We found that two different pHs are required, one for the immobilisation of the antibodies on gold (pH 5.5) and a different one for optimal interaction between the sample and the antibody layer (pH 7.0). Finally, we briefly demonstrate the application of the sensor to real saliva samples, both mucin-less and mucin-containing, evaluating the potential of the sensor to discriminate between healthy and ill.     

Determination of Nucleic Acid at Nanogram Levels with Manganese-Tetrasulfonatophthalocyanine Sensitized by Cetyltrimethylammonium Bromide Using a Resonance Light-Scattering Technique

This is the first report on the determination of deoxyribonucleic acids based on the enhancement of resonance light-scattering (RLS) of manganese-tetrasulfonatophthalocyanine (MnTSPc) by DNA and cetyltrimethylammonium bromide (CTMAB). Under the conditions of pH 10.6011.85 and an ionic strength of 0.01molL^–1 (NaCl), the interaction of MnTSPc with DNA sensitized by CTMAB results in enhanced RLS signals at 314nm, 346.2nm, 452.6nm and 494.4nm in the enhanced regions. It was found that the enhanced RLS intensity at 346.2nm was proportional to the concentration of DNA in the suitable ranges. The limits of detection were 11.3ngmL^–1 for fsDNA and 22.9ngmL^–1 for ctDNA when the concentration of MnTSPc was chosen to be 2.0×10^–6molL^–1. This method was simple, rapid, and the dye (MnTSPc) was easily synthetized, inexpensive, and stable. We applied it to the determination of DNA in synthetic samples with satisfactory results.Received November 4, 2002; accepted January 18, 2003 Published online July 16, 2003     

Determination of proteins in human serum by combination of flow injection sampling with resonance light scattering technique

A flow injection method combined with Resonance light scattering detection was developed for the determination of protein concentration in human serum samples. This method is based on the enhanced RLS signals of protein binding with the dye acid chrome blue K. The enhanced RLS intensities at 264 nm, in an acidic aqueous solution, were proportional to the protein concentration over the range of 2.0–40.0 μg·mL⁻¹ for human serum albumin (HSA) and the limit of detection (3σ) is 85 ng·mL⁻¹. This method was successfully applied to the quantification of total proteins in human serum samples. The maximum relative standard deviation is less than 2% and the recovery is between 97 and 103% for the standard addition method. The sample throughput was 60 h⁻¹.     

Seeking connectivity between engineered proteins and transducers: connection for glutathione S-transferase fusion proteins on surface plasmon resonance devices

The affinity bond between glutathione (GSH) and glutathione S-transferase (GST) is exploited as a means for ‘connectivity’ for engineered proteins at a surface plasmon resonance (SPR) surface. If the protein of interest is recombinantly fused to GST the resulting fusion protein can be linked specifically to GSH self-assembled on a gold surface. Classical self-assembly and the potential assisted self-assembly were compared. The classical method produced unstable layers. Applying a potential during the assembly process significantly improved stability and reproducibility. Suitable GSH layers could be deposited at potentials >0.2 V versus Ag/AgCl, where the reductive desorption showed no desorption peaks between −0.85 and −1.1 V. The GSH-functionalised surface was tested for applicability with the plant cyclin-dependent kinase (CDK) Cdc2aAm recombinantly fused to GST. The fusion protein maintained both the affinity for glutathione and the activity for cyclin binding of its parent proteins. SPR signals due to the interaction of Cdc2aAm with a cyclin-binding-site specific antibody were confirmed by ELISA. In this instance, this test system opens up the possibility of studying the cell cycle machinery, but more widely the issues concerned with maintaining the correct conformations of proteins to achieve protein arrays can be developed from this method.     

FIRE: predicting the spatial proximity of protein residues from 3D NOESY–HSQC

 We address the problem of the prediction of residue spatial proximity in a protein, through the automatic processing of a 3D ¹⁵N NOESY–HSQC. The spatial distance between residues is estimated from a spectral match value calculated using a comparison of the resonances involving the amide hydrogens. The method is shown to provide a good estimation of a large number of residue spatial proximities, in the case of two experimental 3D spectra, recorded on proteins of α and β secondary structures. It is tested on simulated data sets against the protein size, secondary structure and the quality of the signal. More than 70% of the sequential assignment is correctly predicted, and the prediction is better for the α than for the β secondary structure. The medium- and long-range correlations seem equally well predicted for all the secondary structures. The efficiency of the method is compared to a previously proposed spectral correlation approach. Received: 5 July 2000 / Accepted: 8 September 2000 / Published online: 19 January 2001     

Determination of nucleic acids at nanogram levels using the resonance light scattering technique with 3,3′-dichlorobenzidine

A novel method for the determination of nucleic acid at nanogram levels was developed based on the measurement of resonance light scattering (RLS) signals of 3,3′-dichlorobenzidine (DCB). In the Britton-Robinson buffer (pH 2.21), the weak light scattering of DCB was greatly enhanced by addition of calf thymus DNA (ctDNA), the maximum RLS peak is at 346 nm and the enhanced intensity of RLS is in proportion to the concentration of ctDNA. The linear range is 0.05–5 μg mL⁻¹ for ctDNA, and the detection limit is 14 ng mL⁻¹ (3σ). DNA in synthetic samples was analyzed with satisfactory results.     

Effective modeling of intrinsic and environmental effects on the structure and electron plaramagnetic resonance parameters of nitroxides by an integrated quantum mechanical/molecular mechanics/polarizable continuum model approach

 Experimental and density functional theory geometries have been used to extend the AMBER force field to nitroxides. An optimum set of transferable atomic charges for the calculation of electrostatic interactions both in vacuo and in aqueous solution has been obtained by averaging the charges obtained by a restrained electrostatic potential fitting of representative compounds. Besides reliable structural data, our implementation allows the computation of accurate spectromagnetic properties by single-point B3LYP computations on geometries optimized at the AMBER level. Solvent shifts in aqueous solution can be reproduced quantitatively by a mixed model in which specific solvent effects are described by two water molecules strongly coordinated to the nitroxide oxygen, while bulk effects are described by the polarizable continuum model. Received: 17 September 1999 / Accepted: 3 February 2000 / Published online: 29 June 2000     

Electronic Circular Dichroism-Circularly Polarized Raman (eCP-Raman): A New Form of Chiral Raman Spectroscopy

This Concept article summarizes recent work on the development of a new form of chiral Raman spectroscopy, e CP-Raman, which combines two spectroscopies: electronic circular dichroism (ECD) and circularly polarized Raman (CP-Raman). First, some puzzling observations while carrying out Raman optical activity (ROA) measurements of several transition metal complexes under resonance are described, as well as the search for the mechanisms responsible. Then an equation for quantifying the e CP-Raman contribution is presented, followed by several examples of how e CP-Raman influences the I_R−I_L spectra of achiral and chiral solvent molecules and of a number of chiral solutes under resonance. The conditions to extract resonance ROA, when the e CP-Raman contribution is minimized, are also discussed. Finally, we comment on the potential applications of e CP-Raman.     

A Highly Sensitive Resonance Rayleigh Scattering Method for the Determination of Vitamin B1 with Gold Nanoparticles Probe

In weakly acidic buffer medium, vitamin B₁ (VB₁) interacts with gold nanoparticles to form a binding product, which resulted in a significant enhancement of resonance Rayleigh scattering (RRS) intensity and the appearance of a new RRS spectrum. The maximum RRS peak was at 368 nm, and there are three smaller scattering peaks that were at 284 nm, 440 nm and 495 nm, respectively. The enhanced RRS intensity (ΔI) was directly proportional to the concentration of VB₁ in the range of 0–2.8 × 10⁻⁷ mol L⁻¹. The method had high sensitivity and its detection limit (3σ) was 0.9 ng mL⁻¹. The optimum conditions and the influencing factors have been investigated. The method had good selectivity, which could be observed from the influence of coexisting substances. A sensitive, simple and fast RRS method for the determination of VB₁ with gold nanoparticle probe has been developed. In addition, the reasons for RRS enhancement were discussed.     

Sensitive Determination of DNA by Resonance Light Scattering with Pentamethoxyl Red

A novel sensitive method for the determination of nucleic acid (DNA) using the resonance light scattering (RLS) spectra of pentamethoxyl red has been developed. It is based on the effects on the resonance light scattering of Pentamethoxyl Red. The effective factors and the optimum conditions were studied, and the enhanced intensity of RLS is in proportion to the concentration of nucleic acids in the range of 0–2.54 µg mL⁻¹ for ct-DNA, 0–4.54 µg mL⁻¹ for hs-DNA. The limits of detection are 1.1 and 2.1 ng mL⁻¹, respectively. Most foreign substances do not interfere in the determination, and the method has good selectivity and high sensitivity. It has been applied to the determination of DNA in synthetic samples and in real samples with satisfactory results.     

CH2Cl2分子中CH的摇摆振动与伸缩和弯曲振动的耦合

Infrared absorption spectra of gaseous CH2Cl2 in the regions of 1200-12000 cm-1 were measured using a Bruker IFS 120HR Fourier transform spectrometer in conjunction with a multipass cell. 47 vibrational levels of overtone and combinational spectral lines of the CH stretching (v1, v6), bending (v2), and rocking(v8) modes were analyzed and assigned. Utilizing the normal mode model and considering the coupling among CH stretching, bending and rocking vibrations, values of the harmonic frequency ωi, the anharmonic constant xij, and the coefficients of Fermi and the Darling-Dennison resonances of v1, v6, v2 and v8 modes were also determined from experimental spectral data with nonlinear least-square fitting. These spectral constants reproduced the experimental levels very well. These results showed that Fermi resonance between CH stretching and rocking vibrations (ki88=-254.63 cm-1) is stronger than that between CH stretching and bending vibrations (k122 = 54.87 cm-1); and that Darling-Dennison resonances between CH stretching and bending vibrations (k1166=-215.28 cm-1) is also much stronger than that between CH bending and rocking vibrations (k2288=-5.72 cm-1).     

金纳米粒子共振散射与共振吸收的关系

金纳米粒子共振散射与共振吸收的关系;金纳米粒子;共振散射;共振吸收     

Simple and sensitive assay for nucleic acids by use of the resonance light-scattering technique with the anionic dye methyl blue in the presence of cetyltrimethylammonium bromide

This is the first report on the determination of nucleic acids based on the enhancement of resonance light scattering (RLS) of the anionic dye methyl blue (MB) in the presence of cetyltrimethylammonium bromide (CTMAB). In tris(hydroxymethyl) aminomethane buffer of pH 9.0, MB and nucleic acids react with CTMAB to form large particles of complex, which results in strong enhanced RLS signals characterized by three peaks at 334 nm, 393.5 nm and 548 nm. Mechanistic studies show that the enhanced RLS stems from the aggregation of MB on nucleic acids through the bridged and synergistic effect of CTMAB. With the enhanced RLS signals at the best wavelength at 334 nm, the enhanced RLS intensity is proportional to the concentration of nucleic acids in a wide range. The lowest limit of determination was 2.1 ng mL⁻¹, three synthetic samples were analyzed satisfactorily.     

基于共振能量转移的生物传感器在食品安全检测中的应用研究进展

共振能量转移(Resonance energy transfer,RET)是一种发生在供体和受体之间的非辐射能量转移过程。RET的能量转移效率对供体和受体间的距离变化非常敏感,可被用于开发新型的光学生物传感器。与传统光学生物传感器相比,基于RET的生物传感器无需洗涤及分离过量标记物等步骤,可大幅简化检测流程。因RET具有灵敏度高、操作简便及速度快等优点,近年来,在医学诊断、生命科学研究、环境监控以及食品安全检测等领域备受关注。该文根据能量供体的不同,将RET分为3种类型：荧光共振能量转移(Fluorescence resonance energy transfer,FRET)、生物发光共振能量转移(Bioluminescence resonance energy transfer,BRET)和化学发光共振能量转移(Chemiluminescence resonance energy transfer,CRET)。并分别对基于上述3种RET类型的生物传感器在食品安全检测中的应用研究进展进行了综述,同时对其应用前景和发展趋势进行了展望。