Matrix effects in immunobiosensor determination of clenbuterol in urine and serum

The construction of immunochemical inhibition assays for beta-agonist and hormone residues have previously been described. In the present work the beta-agonist assay was further optimised for application to biological samples, using urine as the main model matrix. Matrix interferences with the antigen-antibody interaction and non-specific binding (NSB) of matrix components to the sensor surface were systematically studied. A full factorial design experiment was employed for evaluating the effects of assay buffer composition. In addition, the influence of antibody concentration and sample dilution on the matrix background was investigated. NSB from urine was highly affected by buffer pH and salt concentration, while buffer composition had little effect on matrix interferences with the antigen-antibody interaction. Ultra-filtration efficiently prevented NSB from urine and serum samples. Increased antibody dilution reduced the matrix background while sample dilution had an opposite effect.     

Direct identification of all oncogenic mutants in KRAS exon 1 by cycling temperature capillary electrophoresis

Over the past few decades, advances in genetics and molecular biology have revolutionized our understanding of cancer initiation and progression. Molecular progression models outlining genetic events have been developed for many solid tumors, including colon cancer. Previous reports in the literature have shown a relationship between different KRAS mutations and prognosis and response to medical treatment in colon cancer patients. Furthermore, the presence of a mutated KRAS has been correlated with different clinicopathological variables including age and gender of patients and tumor location. To our knowledge, few institutions screen for KRAS mutations on regular basis in colon cancer patients despite such evidence that knowledge of KRAS exon 1 status is informative. Here, we report on a mutation analysis method adapted to a 96-capillary electrophoresis instrument that allows identification of all 12 oncogenic mutations in KRAS exon 1 under denaturing conditions. To determine the optimal parameters, a series of DNA constructs generated by site-directed mutagenesis was analyzed and the migration times of all mutant peaks were measured. A classification tree was then made based on the differences in migration time between the mutants and an internal standard. A randomized series of 500 samples constructed with mutagenesis as well as 60 blind samples from sporadic colon carcinomas was analyzed to test the method. No wild-type samples were scored as mutants and all mutants were correctly identified. Post polymerase chain reaction (PCR) analysis time of 96 samples was performed within 40 min.     

Differentiation and quantification of synthetic phosphatidylethanol (PEth) homologues by 1H- and 13C-NMR in polar organic solvents

Various phosphatidylethanol (PEth) derivatives, the corresponding reversed positional isomers (RPI-PEths), lyso-PEth-16:0, and penta-deuterium-labeled PEth analogs (d5-PEths), were synthesized by enzyme-independent synthetic routes. A general solvent system consisting of a mixture of acetone-d6 and methanol-d4 (97:3; v/v) was found to provide a good solubilizing capacity and excellent hydrogen-1 NMR (¹H-NMR) peak resolution of various PEth homologues. Analytical differentiation of PEth from the corresponding RPI-PEth by carbon-13 NMR (¹³C-NMR) was demonstrated by comparison of the ¹³C-NMR signals of the carbonyl groups, the allylic positions, and of the β-carbons. An exemplary stable long-term room temperature, DMSO-d6-based, and proton-sensitive quantitative nuclear magnetic resonance (¹H-qNMR) independently quantified calibrator comprising PEth-16:0/18:1 for liquid chromatography (tandem) mass spectrometry (LC-MS/MS) analytical applications were prepared by employment of sodium dodecyl sulfate (SDS) as a solubilizing additive. In summary, novel hypothetically occurring PEth derivatives, e.g., RPI-PEths, have been independently synthesized with regio- and stereochemical control. Use of polar organic solvents, e.g., mixtures of acetone-d6 and methanol-d4 or DMSO-d6, improves spectral line shapes as compared to traditional hydrophobic solvents and allow for analytical differentiation between closely related PEth derivatives, as well as LC-MS/MS-independent concentration determination of dissolved single species by employment of ¹H-qNMR.

On the influence of the nuclear quadrupole moment on the β-decay of deformed nuclei

It is pointed out that in none of the previous treatments of the quadrupole interaction in β-decay have the relevant boundary conditions on the final state been recognized or observed. The discrepancy between a purely perturbative treatment and some slightly more exact treatments is shown to stem, ultimately, from this circumstance. Similarly, the basic argument behind the prediction of discontinuities in the β-spectra of deformed nuclei is incompatible with the pertaining boundary conditions on the final state. A straightforward adherence to the proper boundary conditions results in discontinuities which, in general, are much smaller than previously anticipated. The failure to see a predicted discontinuity in a reported preliminary experiment on the β-decay of ¹⁷⁰Tm is entirely understandable, without any assumption of some kind of further hindrance in the decay.     

On the theory of scattering by complex potentials

The scattering problem for a non-relativistic spinless particle under the influence of a complex effective potential, which is spherically symmetric and tends to zero faster than 1/r at infinity, is considered. Certain general relations, which illuminate the influence of the imaginary part of the potential on the scattering process, are derived with the use of the expression for the probability current density. The rigorous phase-integral method developed by N. Fröman and P. O. Fröman is used for obtaining an exact, general formula for the scattering matrix, or, equivalently, for the phase shift. The formula is expressed in terms of phase-integral approximations of an arbitrary order and certain quantities defined by convergent series. Estimating the latter quantities and omitting small corrections, an approximate formula is derived for the phase shift, valid for the case that only one complex turning point contributes essentially to the phase shift. Criteria for classifying a scattering problem as such a one-turning-point problem are given. The treatment is made general enough to also cover situations of interest in Regge-pole or complex angular momentum theory.     

On Calculations of Legendre Functions and Associated Legendre Functions of the First Kind of Complex Degree

Formulas for calculating Legendre functions and associated Legendre functions of the first kind of complex degree using an Ermakov—Lewis invariant are presented. These formulas are straight-forward to implement numerically and are motivated by the lack of computational routines in standard university tools like those of MatLab and Maple. Angular waves propagating in opposite directions are also obtained. The results are particularly useful in complex angular momentum theories and nearside/farside analysis of spin-dependent angular scattering from central potentials.     

The HEATOX Project

HEATOX is the acronym for a European Union-funded project entitled Heat-Generated Food Toxicants: Identification, Characterization, and Risk Minimization. Acrylamide will be the main experimental focus, but identification of unknown toxicants in heated carbohydrate-rich foods will also be attempted. The project includes research on formation chemistry, food technology, analytical methods, hazard characterization, and exposure assessment. The results will finally be used in risk assessment and risk management advice.     

DISTRIBUTION OF 5-AMINOLEVULINIC ACID-INDUCED PORPHYRINS IN NODULOULCERATIVE BASAL CELL CARCINOMA

Abstract— Microscopic fluorescence photometry incorporating a light-sensitive thermo-electrically cooled charge-coupled device (CCD) camera was employed to investigate the fluorescence distribution of 5-aminolevulinic acid (ALA)-induced porphyrins in 22 patients with a total number of 52 noduloul-cerative basal cell carcinomas (BCC) after topical ALA application with or without dimethylsulfoxide (DMSO)/ethylenediaminetetraacetic acid (EDTA) or after intravenous administration of ALA. Both localization patterns and amounts of ALA-induced porphyrins in the BCC were studied. The ALA-induced porphyrins were localized only in the superficial layers of the noduloulcerative BCC lesions after topical application of 20% ALA alone for 3 h. However, both the penetration of ALA into deep lesions and the production of the ALA-induced porphyrin fluorescence were increased after topical administration of 20% ALA and 20% DMSO/4% EDTA for 3 h. Prior treatment with 99% DMSO for 15 min further enhanced the ALA penetration into the BCC lesions after topical application of the ALA/DMSO/EDTA mixture and produced more ALA-induced porphyrins by a factor of about three compared with those treated with ALA alone. The penetration of ALA into the deep BCC lesions could also be increased by prolonging the time of topical application of 20% ALA/4% EDTA to 29–48 h (without DMSO). Intravenous injection of ALA led to a more homogeneous distribution of the ALA-derived porphyrins in the whole noduloulcerative BCC lesions.     

Analysis of acrylamide in cooked foods by liquid chromatography tandem mass spectrometry

A method using liquid chromatography tandem mass spectrometry (LC-MS-MS) with electrospray for the analysis of acrylamide in foods is reported. The method comprises the addition of deuterium-labelled acrylamide-d3, extraction with water, mixed mode solid phase extraction, ultrafiltration and a graphitised carbon column for chromatography. The transitions m/z 72 > 55, 72 > 54, 72 > 44, 72 > 27, 72 > 72 and 75 > 58 were recorded in multiple reaction monitoring mode for identification and quantification. In-house validation data for products from potatoes and cereals (30 to 10,000 microg kg(-1)) are presented (accuracy 91 to 102%, relative standard deviation 3 to 21%). Interlaboratory validation data (comparison with gas chromatography mass spectrometry, 25 to 2000 microg kg(-1)) showed excellent results (r2 = 0.998).     

Optimization of the determination of selenium in marine samples by atomic absorption spectrometry: Comparison of a flameless graphite furnace atomic absorption system with a hydride generation atomic absorption system

A comparison has been made between a graphite furnace system based on nickel as a matrix stabilizing metal and an automated hydride generation system with a heated quartz cell. The effect of nickel as a matrix modifier was studied in pure selenite solutions as well as in biological matrixes by different charring temperatures. The suppression effect of different acids on the response of the analyte is reported and discussed. The use of an electrically heated quartz tube as an alternative to the argon hydrogen flame method unproved the selenium determination by hydride generation atomic absorption. The effect of hydrochloric acid to secure quantitative formation of selenium (IV) and the interference of copper in the response measurements have been studied. Further a comparison has been made between three different digestion procedures when the hydride generation atomic absorption system was applied. The results of the graphite furnace atomic absorption and the hydride generation atomic absorption were found to be equally accurate, but the graphite furnace technique gave better reproducibility.