A novel flow injection chemiluminescence method for automated and miniaturized determination of phenols in smoked food samples.

An easily performed fully automated and miniaturized flow injection chemiluminescence (CL) method for determination of phenols in smoked food samples has been proposed. This method includes the ultrasound assisted solid-liquid extraction coupled with gas-diffusion separation of phenols from smoked food sample and analytes absorption into a NaOH solution in a specially designed gas-diffusion cell. The flow system was designed to focus on automation and miniaturization with minimal sample and reagent consumption by inexpensive instrumentation. The luminol - N-bromosuccinimide system in an alkaline medium was used for the CL determination of phenols. The limit of detection of the proposed procedure was 3·10-8·molL-1 (0.01mgkg-1) in terms of phenol. The presented method demonstrated to be a good tool for easy, rapid and cost-effective point-of-need screening phenols in smoked food samples.

Simultaneous determination of iron (II) and ascorbic acid in pharmaceuticas based on flow sandwich technique.

The simple and easy performed flow system based on sandwich technique has been developed for the simultaneous separate determination of iron (II) and ascorbic acid in pharmaceuticals. The implementation of sandwich technique assumed the injection of sample solution between two selective reagents and allowed the carrying out in reaction coil two chemical reactions simultaneously: iron (II) with 1,10-phenanthroline and ascorbic acid with sodium 2,6-dichlorophenolindophenol. For achieving of excellent repeatability and considerable reagent saving the various parameters such as flow rate, sample and reagent volumes, reaction coil length were also optimized. The limits of detection (LODs) obtained by using the developed flow sandwich-type approach were 0.2 mg L(-1) for iron (II) and 0.7 mg L(-1) for ascorbic acid. The suggested approach was validated according to the following parameters: linearity and sensitivity, precision, recoveries and accuracy. The sampling frequency was 41 h(-1).

Role of temperature on protein and mRNA cytochrome P450 3A (CYP3A) isozymes expression and midazolam oxidation by cultured rat precision-cut liver slices.

The cytochrome P450 3A (CYP3A)-mediated midazolam oxidation was studied in rat precision-cut liver slices (PCLS) maintained for 20hr at 4, 20 and 37 degrees, and further incubated for 8hr at 37 degrees. Either at 4 or 20 degrees, midazolam was oxidised by PCLS at similar rates to that observed in freshly cut slices. Moreover, PCLS kept a regioselectivity since 4-hydroxylation was more important than 1'-hydroxylation. Conversely, PCLS totally lost their capacity to oxidise midazolam after 20hr at 37 degrees, and both CYP3A2 protein and mRNA were not detected. CYP3A1 protein was unaffected by a temperature of 37 degrees but its mRNA was totally lost. By blocking transcription with actinomycin D, the decay of both CYP3A mRNAs followed the same profile at either 20 or 37 degrees, indicating that temperature affected the CYP3A2 protein stability. Cell functionality was not involved in such an impairment since the low values of ATP, GSH and protein synthesis rates observed at 4 and 20 degrees were rapidly restored, when PCLS were further incubated at 37 degrees. The use of rat supersomes expressing either CYP3A1 or CYP3A2, strongly supported the hypothesis that 4-hydroxymidazolam was mainly formed by CYP3A2. These results suggest that: (1) CYP3A1 protein is constitutive and largely expressed in rat liver slices; (2) regioselective midazolam oxidation appears to be mainly CYP3A2 dependent; and (3) since CYP3A isoforms have similar half-lives (about 10-14hr), the loss of CYP3A2 protein at 37 degrees might be due to a selective targeting (phosphorylation ?) leading to proteolytic disposal by the proteasome.