Development of a Flow Immunoassay System for the Detection of Salmonella Typhimurium

Abstract

A flow immunoassay system has been developed for the detection of Salmonella typhimurium by immobilization of antibody to Salmonella onto tygon tubing by adsorption and onto polyethylene tubing, by covalent binding. After liquid containing Salmonella cells or Salmonella antigen was allowed to interact with the immobilized. antibody, glucose oxidase labeled antibody to Salmonella was injected to the system. The bound antigen was quantified by electrochemical detection of H₂O₂ produced from the enzymic oxidation of glucose to gluconic acid. In view of the low non-specific binding, sensitivity and reusability, the polyethylene immunoreactor provided a better performance. The system was capable of detecting as low as 10⁵ ? 10⁶ Salmonella cells/ml in less than 10 minutes. The immunoreactor could be regenerated for at least 50 times during one month of the experiment. The detection limit of the tygon immunoreactor was somewhat higher (10⁶ ? 10⁹ cells/ml). Even though the anti-Salmonella antibody was easily immobilized on the tygon tubing, the system was not reusabl and exhibited a very high non-specific binding.     

一种用于测定尼古丁的新型压电石英晶体传感器的研制

本文将含有四苯硼钠的聚氯乙烯(PVC)敏感膜涂渍在压电石英晶体(PQC)表面,制成尼古丁传感器,该传感器在pH 5.0~7.5的范围内,对1.0×10-6~1.0×10-2mol/L的尼古丁水溶液有很好的线性响应,检出限为8.0×10-7mol/L。本法较紫外分光光度法有更宽的线性范围和更低的检出限。     

Piezoelectric Crystal Biosensor System for Detection of Escherichia Coli

Abstract

A piezoelectric crystal biosensor system was applied to the detection of Escherichia coli. the system consists of an oscillator, a frequency counter, a flow cell and a modified piezoelectric crystal. Anti-E. coli antibody is immobilized on the surface of the crystal. It is used as an E. coli detection by measuring its resonant frequency shift due to a mass change caused by specific binding of the micro organisms to the surface. the frequency shift correlates with an E. coli concentration in the range of 10⁶?10⁸ cells·cm^?3. the resonant frequency shift is increased by further treatment to bind micro-particles modified with anti-E. coli antibody. This method allows us to improve the determination limit to 10⁵ cells · cm^?3.     

Special Report Chemical Sensors for In Vivo Monitoring: Clinical Aspects

Abstract

The origins and purpose of a new European Community Concerted Action on “Chemical Sensors for In Vivo Monitoring” are described: the overall aim is to promote the development and application of chemical sensors for in vivo monitoring. A report is presented on the first Workshop Meeting entitled “Clinical Aspects of In Vivo Sensing”, which addressed the question of the analyte to be sensed, specific and general problems of in vivo sensing, and future prospects.     

The Ambiguity of the Viscometric Method for the Deter-Mination of Dextranase Activity

Abstract

In earlier studies it was found that the tissue pellets of Avena seedlings decrease the viscosity of dextran solutions. By the use of direct methods for investigating the breakage of these molecules, it is shown that the lowering of viscosity of dextran solutions under the influence of tissue pellets of Avena coleoptiles is not due to an intramolecular process of molecular breakage (dextranase-like activity) but rather to an intermolecular process of still unknown nature. This conclusion is supported by the finding that solutions of entirely different macromolecules than dextrans are also affected by the tissue pellets. The observed changes in viscosity can therefore not be due to dextranase activity and the viscometric method for the determination of this activity is clearly ambiguous.     

A Direct Spectrophotometric Assay for Pyruvic Oxime Hydrolase

Abstract

A sensitive spectrophotometric method, which directly measures the formation of pyruvic oxime from pyruvate and NH₂OH in the UV, is described for the detection of pyruvic oxime hydrolase activity in biochemical systems. The method was used in an attempt to detect pyruvic oxime hydrolase activity in cell-free extracts from an Alcaligenes sp. which can grow on pyruvic oxime. Although substantial pyruvic oxime + nitrite oxidative activity was detected in cells and cell-free extract of this bacterium, catalysis of pyruvic oxime formation was not observed within the error of the method (～20% of the uncatalyzed rate constant of 1.9x10^?3 s^?1 at pH 7, 22°C). The rate of pyruvic oxime oxidation by cells and cell-free extract was at least 10⁵?10⁶ times greater than the rate of its hydrolysis, thus implying that oxidation of pyruvic oxime need not require prior hydrolysis. The method would appear to be applicable to hydrolases directed toward a variety of oximes.     

Base-Catalyzed Rate and Arrhenius Activation Energy for t-Butyl Isothiocyanate Hydrolysis in Dioxane-H2O and Ethanol-H2O Mixtures by Gas Chromatography

Abstract

N-bonded tertiary-butyl isothiocyanate hydrolyzes via a mechanism that differs from the normal, or S-bonded linkage isomer. The diminutive amount of published kinetic data for the former compound prompted a redetermination of the base-catalyzed rate of hydrolysis of the isothiocyanate in 2% dioxane, ethanol water mixtures at several temperatures using gas-liquid chromatography.     

Influence of Methyl Substitution on mass Spectra of Diphenylmethanes: Analytical Applications

Abstract

In connection with other studies we synthesized and obtained low resolution electron impact spectra of the 2,2′-, 2,3′-, 2,4′-, 3,3′-, 3,4′-, and 4,4′-dimethyl substituted diphenylmethanes. The results suggest that caution should be exercised in using the characteristic even mass rearrangement fragment (m/e = 104) produced by the “ortho-effect” as the only method to distinguish between ortho and non-ortho methyl substituted diphenyl-methanes. The relationship between the stability of the diphenylmethane isomers and the substitution patterns at the meta and para positions does not parallel that observed for the dimethyl substituted biphenyls.     

Fluorescence Behavior of Phenolic Compounds in Solutions Containing Micelles

Abstract

The effects of micelles on the fluorescence detection of phenolic compounds are described. Micelles were found to enhance the fluorescence of the drugs apomorphine (APO), and N-n-propylnorapomorphine (NPA), and a series of model hydroxybiphenyls. The fluorescence enhancements were pH dependent and biphasic behavior in hexadecyltrimethylamnonium chloride was observed for APO and NPA. Multiple equilibria and formation of zwitterions of the phenolic aporphines in the micellar environment is proposed to explain this behavior. Micellar fluorescent enhancements should be useful in improving the sensitivity of fluorometric determinations of polar compounds.     

Pyrocatechol-1-Aldehyde 2-Benzothiazolylhydrazone As Reagent for the Spectrofluorimetric Determination of Nanogram Amounts of Gallium in Urine and Blood Serum

Abstract

A method is developed for the spectrofluorimetric determination of 1–80 ng.ml^?1 of gallium with pyrocatechol-1-aldehyde 2-benzothiazolylhydrazone, in a 50% (v/v) DMSO-Water medium at apparent pH 4.0 (monochloracetic/monochloracetate buffer). Λ_ex = 400nm, Λ_em = 504 nm (corrected). Interferences have been evaluated and the method applied to the determination of gallium in human urine and blood serum samples.     

Spectrophotometric Determination of Manganese with 4(5)-Imidazolealdoxime (Imalox)

Abstract

A method is described for the spectrophotometric determination of manganese by means of its complex with 4(5)-imidazolealdoxime, IMALOX, formed in alkaline medium. The colour system has its absorption maximum at 350 nm and obeys Beer's law over the range 0.8–8.0 μg of Mn per ml. The optimum range is 1–5 μg m^?1. The molar absorptivity is 7850 1 mol^?1 cm^?1. Relatively few ions interfere, and these can be masked with cyanide, tartrate and NTA. The experimental results have been critically analyzed and a comparison with the main spectrophotometric reagents for manganese is presented.     

A Fluorometric Assay of L-Aspartate: 2-oxoglutarate Aminotransferase in Human Serum with N-(9-Acridinyl) maleimide

Abstract

A new assay of L-aspartate:2-oxoglutarate aminotransferase (EC 2, 6, 1, 1) (glutamic-oxaloacetic transaminase: GOT) is reported. Substrate L-aspartate was replaced with L-cysteine sulfinic acid (CSA) which yields sulfite and pyruvate. The enzyme activity was determined by measuring the sulfite with the aid of a fluorescent reagent, N-(9-acridinyl)maleimide (NAM). The method presents a extremely sensitive GOT assay in the range of 4 to 140 U/l using less than 10 μl of human serum.     

Quantitative Determination of Nortriptyline And Desmethylnortriptyline In Human Plasma By Combined Gas Ciiromatography - Mass Spectrometry

Abstract

A quantitative method has been developed for the determination of the tricyclic antidepressant drug nortriptyline in plasma, utilizing a combined technique of gas chromatography - mass spectrometry called mass fragmentography. A metabolite, desmethylnortriptyline, may be analyzed in the same run. The compounds are first converted to their heptafluorobutyranide derivatives. A complete chromatogram takes less than five minutes. Quantitive determinations under the present conditions are possible down to a few nanograms of nortriptyline per milliliter. There is a good correlation between the new method and that previously used which was based upon in vitro ³ 11-acetylation labelling.     

Rapid determination of phenoxy acid residues in rice by modified QuEChERS extraction and liquid chromatography-tandem mass spectrometry

A new method for the analysis of phenoxy acid herbicide residues in rice, based on the use of liquid extraction/partition and dispersive solid phase extraction (dispersive-SPE) followed by ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS), is reported. 5% (v/v) formic acid in acetonitrile as the extraction solvent and inclusion of citrate buffer helped partitioning of all the analytes into the acetonitrile phase. The extract was then cleaned up by dispersive-SPE using C18 and alumina neutral as selective sorbents. Further optimization of sample preparation and determination allowed recoveries of between 45 and 104% for all 13 phenoxy acid herbicides with RSD values lower than 13.3% at 5.0 μg kg⁻¹ concentration level. Limit of detections (LODs) of 0.5 μg kg⁻¹ or below were attained for all 13 phenoxy acids. Quantitative analysis was done in the multiple-reaction monitoring (MRM) mode using two combinations of selected precursor ion and product ion transition for each compound. This developed method produced relatively higher recoveries of the acid herbicides with a smaller range of variation and less susceptibility to matrix effects, than the original QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method.     

Spectrophotometric Determination of Nickel(II) with O-hydroxyhydroquinonephthalein in the Mixed Micellar Media of Zephiramine and Tween 20

Abstract

The effect of mixed surfactants on the color reaction between nickel(II) and o-hydroxyhydroquinonephthalein(Qnph) was examined in weakly basic media. The color development of the metal complex in the presence of cationic and nonionic surfactants (mixed micellar media of Zephiramine (Zp) and Tween 20) was very stable and exhibited a high and reproducible absorbance. The calibration curve was linear in the ranges of 0 – 7.0 μg nickel(I1) in a final volume of 10 ml at 545 or 536 nm. The method developed does not require an organic solvent extraction step. The apparent molar absorptivity and Sandell sensitivity were 1.26 × 10⁵ 1 per mol per crn and 0.0045 μg per cmL nickel(II), respectively, at 545 nm.     

Crystal structure and conformation of 10-aryl-bilatrienes-abc

Summary Crystal structure determinations of etiobiliverdin-IV (1) and of twomeso-substituted arylbilatrienes-abc (the 2-imidazolylderivative2 and the 4-nitrophenylderivative3) are reported. All three molecules have aZ,Z,Z configuration and occur in the crystal in anall-syn conformation, which results in a helical overall molecular shape. As a consequence of the steric repulsion between the aryl substituent and the two methyl groups on positions 8 and 12, the pitch and the curvature of the helix are larger in the twomeso-substituted derivatives2 and3 compared to1. In each structure, the packing is dominated by the formation of centrosymmetric, S-shaped dimers via two hydrogen bonds between the pyrrolic H-atom and the carbonyl oxygen of ring A of two molecules related by the operation of an inversion center.

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Kristallstruktur und Konformation von 10-Aryl-bilatrienen-abc

Zusammenfassung Es wird über Kristallstrukturbestimmungen von Aetiobiliverdin-IV (1) sowie zweiermeso-substituierter Arylbilatriene-abc (dem 2-Imidazolylderivat2 und dem 4-Nitrophenylderivat3) berichtet. Alle drei Moleküle liegen im Kristall in einerZ,Z,Z Konfiguration vor und nehmen eineall-syn-Konformation ein, woraus sich eine helikale Molekülgestalt ergibt. Als Folge der sterischen Abstoßung zwischen dem Arylsubstituenten und den beiden Methylgruppen an den Positionen 8 und 12 sind die Ganghöhe und die Krümmung der Helix in den beidenmeso-substituierten Derivaten2 und3 gegenüber1 vergrößert. Die Kristallpackung wird in den drei Kristallstrukturen bestimmt durch die Ausbildung S-förmiger, zentrosymmetrischer Dimeren, die durch zwei intermolekulare Wasserstoffbrücken zwischen dem pyrrolischen Wasserstoffatom und dem Carbonylsauerstoff von Ring A zweier zentro-symmetrieäquivalenter Moleküle zusammengehalten werden.

     

Scattering from concentration fluctuations in polymer blends: A monte carlo investigation

The collective scattering function S_coll( ), which describes light (neutron-, x-ray) scattering under wavevector , is obtained from Monte Carlo simulations for a symmetrical polymer mixture. The polymers are modelled by self-avoiding walks ofN _A=N_B=N steps on a simple cubic lattice, where a fraction _V of sites is left vacant, and an attractive energy occurs if two neighboring sites are taken by the same kind of monomer. Spinodal curves are estimated from linear extrapolation of S _coll ^–1 (0) vs./k _B T, whereT is the temperature. Also the single chain structure factor is obtained and the de Gennes random phase approximation (RPA) can thus be tested. Unexpectedly, strong deviations are found if one species is very dilute. The estimation of an effective Flory-Huggins-parameter from scattering data is also discussed.Contributed paper delivered at the Tagung der Deutschen Physikalischen Gesellschaft, Fachausschuß Polymerphysik, Hamburg, March 14–16, 1987.     

Mild and Efficient Method for Synthesis of Eaminones Using Ytterbium Triflate as Catalyst

A mild and efficient procedure for synthesis of β-enaminones by the condensation of β-dicarbonyl compounds and amines using ytterbium triflate [Yb(OTf)₃] as catalyst is described. The catalyst can be easily recovered and reused without loss of activity.     

A Baylis-Hillman/ozonolysis route towards (±) 4,5-dihydroxy-2,3-pentanedione (DPD) and analogues

The Baylis-Hillman reaction between 2-(tert-butyldimethylsilyloxy)ethanal and 3-buten-2-one followed by desilylation gave rise to the corresponding α-methylene-β,γ-dihydroxy ketone further converted by reductive ozonolysis of the carbon-carbon double bond into racemic 4,5-dihydroxy-2,3-pentanedione (DPD), a significant molecule in bacterial cell-cell communication systems. The same sequence applied to other substrates allowed the preparation of chain elongated analogues and 5-O-acylated derivatives of DPD.