The application of strongly oxidizing agents in flow injection analysis : Part 3. Cobalt(III)

The application of cobalt(III) as a powerful oxidizing agent in flow injection analysis is described. Cobalt(III) is electrochemically generated in the flowing system at a working electrode consisting of a packed bed of gold powder. Depending on the analyte, spectrophotometric detection is at 605 nm or 320 nm. For the oxidizable organic and inorganic substances tested, linear responses are usually obtained in ranges around 10^?3 mol l^?1.     

A BIOCHEMICAL SCREENING TEST FOR THE PHOTOSENSITIZING POTENTIAL OF DRUGS AND DISINFECTANTS

Abstract— A series of photosensitizers known to cause photodermatitis were tested with respect to photosensitizing properties. Irradiation of these sensitizers together with unsaturated fatty acids or amino acids showed that few of them can be transformed by most of the photosensitizers. SH groups can. however, be destroyed by most of the photosensitizers known in dermatological practice. At low photosensitizer-to-SH group ratios, GSSG is formed; at higher ratios, other transformations of GSH predominates. The reactivity of GSH with ninhydrin is decreased after irradiation with a photo-sensitizer. Glutathione seems to be a biomolecule that is suitably sensitive to photosensitized modification and thus suitable for evaluation of any photosensitizing activity that new drugs may have as an unwanted side effect.     

Determination of molar absorptivity coefficients for major type-B trichothecenes and certification of calibrators for deoxynivalenol and nivalenol

This paper presents results from the European Commission-funded project Doncalibrant, the objective of which was to produce calibrators with certified mass fractions of the Fusarium toxins deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-Ac-DON), 15-acetyldeoxynivalenol (15-Ac-DON), and nivalenol (NIV), in acetonitrile. The calibrators, available in ampoules, were sufficiently homogeneous, with between-bottle variations (s _bb) of less than 2%. Long-term stability studies performed at four different temperatures between −18 and 40 °C revealed no significant negative trends (at a confidence level of 95%). Molar absorptivity coefficients (in L mol⁻¹ cm⁻¹) were determined for all four toxins (DON: 6805 ± 126, NIV: 6955 ± 205, 3-Ac-DON: 6983 ± 141, 15-Ac-DON: 6935 ± 142) on the basis of a mini-interlaboratory exercise. The overall uncertainty of the calibrators’ target values for DON and NIV were evaluated on the basis of gravimetric preparation data and include uncertainty contributions from possible heterogeneity, storage, and transport. The Doncalibrant project resulted in the production of calibrators for DON (IRMM-315) and NIV (IRMM-316) in acetonitrile with certified mass fractions of 25.1 ± 1.2 μg g⁻¹ and 24.0 ± 1.1 μg g⁻¹, respectively. Both CRMs became commercially available from the Institute for Reference Materials and Measurements (IRMM, Geel, Belgium) at the beginning of 2007.     

Melanin content of cultured human melanocytes and UV-induced cytotoxicity

Cultured melanocytes originating from persons with different skin phototypes were utilized for measurement of endonuclease sensitive sites induced by UVB and the determination of cell survival after UVA or UVB irradiation. During culture, the melanocytes largely maintained their phenotypic characteristics according to their original skin phototype. Total melanin concentrations were 4.9 times higher in the darker skin phototype (IV-VI) melanocytes when compared to the cells from lighter skin phototypes (I-III). Also phaeomelanin contents were higher (2.5 times) in the skin phototype (IV-VI) melanocytes which implies that the cells from light skin types contain less melanin, but a relatively high proportion of phaeomelanin. After UVB irradiation a stronger induction of endonuclease sensitive sites was found for melanocytes with a lower level of total melanin and a high content of pheomelanin. By measuring the clone forming ability in different melanocyte cultures after UVB irradiation, significant better survival was found in case of the cells with the higher melanin content. Despite the large variations in melanin content, no significant difference in survival after UVA irradiation could be demonstrated in this way. Our results suggest a protective effect of melanin for UVB and indicate the importance of the measurements of melanin content and composition when different parameters of UV-induced damage are studied in melanin producing cells.     

The application of strongly oxidizing agents in flow injection analysis : Part 2. Manganese(III)

The application of manganese(III) as a powerful oxidizing agent in flow injction analysis is described. Manganese(III) is generated electrochemically in the flowing system at a working electrode consisting of a packed bed of gold powder. Spectrophotometric detection is used at 490 nm, where manganese(III) in sulphuric acid solution absorbs strongly. Undr the experimental conditions, the generation of manganese(III) can be accompanied by generation of manganese(IV) and permanganate; manganese(III) alone can be generated by a proper selection of the generating current and the flow rate. Results are presented for the determinatin of various organic and inorganic substances by means of manganese(III), usually at concentrations in the 10^?4—10^? mol l^?1 range. Unlike permanganate and manganese(IV), manganese(III) does not react with chloride, so that oxidizable compounds can be determined in the presence of large amounts of this species.     

PHOTOCHEMICAL DAMAGE TO DNA TREATED WITH CHLORPROMAZINE AND NEAR UV RADIATION UNDER AEROBIC AND ANAEROBIC CONDITIONS

Double stranded salmon sperm DNA in a chlorpromazine (CPZ) solution is damaged when irradiated with near UV light. The damage of irradiated DNA can be estimated by measuring the increase in extinction at 260 nm following incubation at 60°C of the DNA with formaldehyde. Moreover, DNA irradiated in the presence of CPZ or kept in the dark separate quite differently in an aqueous polymer two-phase system. DNA irradiated in the presence of CPZ seemed to be susceptible to digestion by endonuclease S1, while the endonuclease of Neurospora crassa could not digest this DNA. Irradiation under aerobic conditions seemed to be less disastrous for DNA than under anaerobic conditions.     

The application of strongly reducing agents in flow injection analysis : Part 5. Chromium(II) and vanadium(II) in acidic medium

Chromium(II) and vanadium(II) in acidic medium are applied as powerful reducing agents in flow injection analysis. Detection is done amperometrically. For the determination of nitrite with chromium(II), the limit of determination is 5 × 10^?6 mol l^?1 with a linear range up to 7.5 × 10^?5 mol l^?1, similar to the case of uranium(III). Vanadium(II) is less suitable for the determination of nitrite. Nitrate, hydroxylamine and hydrazine could not be determined with these reagents.     

Interference of flavonoids with enzymatic assays for the determination of free fatty acid and triglyceride levels

Flavonoids are bioactive food compounds with potential lipid-lowering effects. Commercially available enzymatic assays are widely used to determine free fatty acid (FFA) and triglyceride (TG) levels both in vivo in plasma or serum and in vitro in cell culture medium or cell lysate. However, we have observed that various flavonoids interfere with peroxidases used in these enzymatic assays, resulting in incorrect lower FFA and TG levels than actually present. Furthermore, addition of isorhamnetin or the major metabolite of the flavonoid quercetin in human and rat plasma, quercetin-3-O-glucuronide, to murine serum also resulted in a significant reduction of the detected TG levels, while a trend was seen for FFA levels. It is concluded that when applying these assays, vigilance is needed and alternative analytical methods, directly assessing FFA or TG levels, should be used for studying the biological effects of flavonoids on FFA and TG levels.     

Results of the second inventory on quality assurance/quality control (QA/QC) and good laboratory practice (GLP) for European Union National Reference Laboratories for residue analyses.

In February 1996 the "Second inventory on QA/QC and GLP for EU-NRLs" was submitted to all European Union National Reference Laboratories (EU-NRLs) for residue analyses. This second inventory was a follow up on the "Inventory on QA/QC" (1993) which was used for the organisation of the first workshop on "GLP for EC-NRLs". In February 1997 a response on the second inventory of 100% was achieved. From the completed inventories it became evident that almost all EU-NRLs for residue analyses now have a described quality program based on an international standard. However only 33% of the EU-NRLs are officially accredited, certified or in compliance with good laboratory practice (GLP) principles. Most of the accredited EU-NRLs have a quality program based on the European Standard EN45001. Seven of the EU-NRLs still do not have a described quality program or have not appointed a quality officer and for 12 of the EU-NRLs the independence of the QA officer is not formally arranged. Only about 50% of the EU-NRLs have a standard operating protocol (SOP) to handle complaints. Almost all of the EU-NRLs have SOPs available and a system to control them. In comparison with the results of the first inventory it is evident that most of the EU-NRLs have made considerable progress in the implementation of quality systems. However it is also evident that a substantial number of EU-NRLs still lack some critical QA facilities. The new deadline for the full implementation of all relevant QA facilities is January 2002.