Single-step microwave digestion of food and biological samples for the quantitative conversion of Se into the +4 oxidation state.

An improved single step microwave digestion procedure is described for providing the fast and easy exhaustive mineralisation of biological samples concomitantly with the quantitative conversion of any type of selenium compounds into Se(IV). In such a way, digested samples are directly suitable for the subsequent Se analysis at trace and ultratrace levels by both spectrometric methods such as HG-ICP-MS or HG-ICP-OES and differential pulse cathodic stripping voltammetry (DPCSV). It is based on the use, under suitably optimised microwave irradiation conditions, of a digestion mixture with a carefully tailored composition such that its redox potential is made lower than that allowing Se(IV) to be oxidized to Se(VI), but high enough to permit total destruction of biological or, in general, organic matrices. It consists of a nitric acid (65%, w/w) and hydrogen peroxide (30%, w/w) mixture in a volume ratio 5:1, frequently adopted for the mineralisation of organic and biological samples, but added simply with 0.25 g mL(-1) of NaCl. Successful application of the procedure, in terms of both repeatability and accuracy, to the quantification of selenium by the instrumental methods above in standard compounds and in a certified biological sample proved its good performance. The application to the Se determination in human blood plasma and in a wide variety of foods is also reported.

Demonstration of labeless detection of food pathogens using electrochemical redox probe and screen printed gold electrodes.

The demonstration of a labeless immunosensor for the detection of pathogenic bacteria using screen printed gold electrodes (SPGEs) and a potassium hexacyanoferrate(II) redox probe is reported. Gold electrodes were produced using screen printing and the gold surfaces were modified by a thiol based self assembled monolayer (SAM) to facilitate antibody immobilisation. SAMs based on the use of thioctic acid (TA), mercaptopropionic acid (MPA) and mercaptoundecanoic acid (MUA) were evaluated. Following antibody immobilisation via the optimum SAM, the redox behaviour and diffusion co-efficient (D) of the potassium hexacyanoferrate(II) probe was monitored in the absence and presence of analyte. In the presence of analyte, a change in the apparent diffusion co-efficient of the redox probe was observed, attributable to impedance of the diffusion of redox electrons to the electrode surface due to the formation of the antibody-bacteria immunocomplex. No change in the diffusion co-efficient was observed when a non-specific antibody (mouse IgG) was immobilised and antigen added. The system has been demonstrated with Listeria monocytogenes and Bacillus cereus.

Human cytomegalovirus detection by a quartz crystal microbalance immunosensor.

A piezoelectric affinity sensor has been developed to detect distinctive antigens of the human cytomegalovirus. Either the specific antibodies or the antigen were immobilized on the gold electrode. To develop a rapid immunoassay, various assay formats were tested in relation with the different antigen composition. First, a direct assay was carried out immobilizing the specific antibody on the crystal surface by passive adsorption. Next, Protein A, thiol/poly L-lysine mixed self-assembled monolayers were tested as methods of gold modification. A competitive format was exploited by immobilization of the antigen onto the crystal activated by SAM and poly L-lysine. This procedure yielded a preliminary calibration curve. A linear range between 2.5 and 5 µg/ml of gB epitope in solution and a detection limit of 1 µg/ml were measured.