THE α-OXIDASE SYSTEM OF YOUNG PEA LEAVES (Pisum sativum) AS GENERATOR OF ELECTRONICALLY EXCITED STATES. EXCITATION IN THE DARK UNDER NATURAL CONDITIONS

Abstract. The aerobic oxidation of saturated long chain fatty acids to the lower aldehyde and CO₂ catalyzed by the α-oxidase system of young leaves from germinating Pisum sativum results in concomitant excitation of the chloroplasts or/and fractions present in the α-oxidase preparation. The excitation is attested to by both chlorophyll emission and Hill activity. This is the first case of photobiochemistry without light within a natural system.     

Analysis of gadobenate dimeglumine by capillary zone electrophoresis coupled with electrospray-mass spectrometry

Highly reliable and accurate analytical methods are needed for the determination of magnetic resonance imaging (MRI) contrast agents in complex matrices of clinical interest. We demonstrate the reliability of capillary zone electrophoresis (CZE) coupled with electrospray ionization-mass spectrometry (ESI-MS) for the analysis of MultiHance (gadobenate dimeglumine), a gadolinium-based MRI agent. A sheath liquid interface connected the CE system with an electrospray mass spectrometer equipped with an ion-trap analyzer. CZE with ultraviolet (CZE-UV) and with mass detection (CZE-MS) were compared by analyzing gadobenate dimeglumine and the free ligand diluted in water and in biological fluids (i.e., human serum and urine). The optimization of some relevant CZE-MS parameters was accomplished, like CE buffer composition, sheath liquid composition and flow, and type and length of the separation capillary. CZE-UV was highly influenced by the biological sample components, which hindered a reliable quantification of both gadobenate and free ligand in serum and urine. In CZE-MS, on the other hand, the electrophoretic runs turned out to be independent of the clinical matrices, due to the informative potential and to the selectivity of MS detection.     

Room-temperature liquid phosphorimetry of the aluminium-ferron chelate in micellar media : Determination of aluminium

Aluminium reacts with 7-iodo-8-quinolinol-5-sulphonic acid (Ferron) in cetyltrimethylammonium bromide (CTAB) micelles to form a highly phosphorescent complex at room temperature. Suitable experimental conditions and the phosphorescent characteristics of the complex are described. Comparison with the results obtained for the phosphorescent niobium-Ferron complex in CTAB micelles helps to elucidate the mechanism of this type of phosphorescence. For aluminium the detection limit is 5.4 ng ml^?1; the relative standard deviation is 4.5% for 20 μg Al. The method is applied to aluminium determination in waters and dialysis fluids.     

Determination of average degree of polymerisation and distribution of oligosaccharides in a partially acid-hydrolysed homopolysaccharide: a comparison of four experimental methods applied to mannuronan

The average degree of polymerisation (DP) and distribution of oligosaccharides in partially acid hydrolysed mannuronans were quantitatively evaluated by 1H NMR, electrospray ionisation mass spectrometry (ESI-MS), micellar electrokinetic capillary chromatography with UV detection (MEKC-UV), and high-pressure anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Our investigation shows that 1H NMR, MEKC-UV and, in particular, HPAEC-PAD can be used as quantitative tools to aid the investigation of polysaccharide structure, function and synthesis. For the latter two techniques, especially, this represents a significant new development as it enables calculation of the quantity of individual oligomers of nominal DP by direct analysis of a defined oligomer mixture. Appropriate statistical averages of number and weight distributions were also calculated and found to fit very well to predicted Kuhn distributions that assume random depolymerisation.     

Synthesis and characterization of aromatic polyamides containing alkylphthalimido pendent groups

A series of polyisophthalamides containing pendent phthalimido groups and flexible side spacers were prepared from four novel diacids and three commercial aromatic diamines. These polyamides were prepared in high yields and with high molecular weights by direct polycondensation with triphenyl phosphite and pyridine as condensing agents. The weight‐average and number‐average molecular weights, measured by gel permeation chromatography, were 70,000–137,000 and 47,000–86,000 g/mol, respectively. The novel polyamides were amorphous and readily soluble and showed glass‐transition temperatures of 150–240 °C, as measured by differential scanning calorimetry. Thermogravimetric analysis showed that the 10% weight‐loss temperatures in nitrogen were 355–430 °C, a significant improvement in thermal stability having been observed with the increase in the side‐chain length. A theoretical quantum mechanical study was successfully carried out to explain these results. Flexible and tough films, cast from polymer solutions, showed tensile strengths of 50–125 MPa. © 2002 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 40: 3711–3724, 2002     

Derivative spectrophotometric resolution of mixtures of the food colourants Tartrazine, Amaranth and Curcumin in a micellar medium

A study of the absorption spectra of the food colourants Tartrazine (E-102), Amaranth (E-123) and Curcumin (E-100) in various organic solvents and pH media, have been carried out. In addition, the spectral behaviour in aqueous micellar systems of anionic, cationic and non-ionic surfactants, as well as beta-cyclodextrin, was investigated. Analytical methods are proposed for the determination of the three dyes in mixtures without any prior separation step, by the application of the derivative technique to the normal absorption spectra, based on the use of cetyltrimethylammonium bromide (CTAB) in 0.1 M NaOH, over the concentration range 2-50 mug ml(-1). The methods are applied for determining the three dyes in commercial food products.     

Preparation of trout liver microsomes for iron speciation in P-450 enzymes by AE–FPLC with ICP–(ORS)MS detection

Cytochromes P-450 are members of a superfamily of hemoproteins involved in the oxidative metabolism of various physiological and xenobiotic compounds in eukaryotes and prokaryotes. The multiplicity of this group of enzymes has been widely studied by chromatographic techniques, mainly high-performance liquid chromatography (HPLC). Because these enzymes are membrane-bound proteins, sample preparation for chromatographic separation of P-450 enzymes requires a solubilization step. The sample-preparation procedures are critical, because detergents affect not only the efficiency of protein solubilization but also their further chromatographic resolution. Trout liver microsomes have been taken here as a model sample to investigate iron speciation in cytochrome P-450. Trouts were treated intraperitoneally with -naphthoflavone, a potent inducer of some P-450 enzymes, and a microsomal suspension containing 7.4±0.1 nmol mL^–1 P-450 enzymes was obtained by ultracentrifugation. Lubrol PX was selected as detergent for solubilization, resulting in about 90% solubilization recovery. The solubilized cytochromes P-450 were further separated by AE–FPLC, with UV detection, or coupled to ICP–MS with an octapole reaction system, ICP–(ORS)MS (monitoring Fe signals at masses 54, 56, and 57). A sampling procedure and chromatographic conditions are developed and were successfully applied to iron speciation in trout liver P-450 enzymes. ICP–(ORS)MS detection of P-450 enzymes is Fe-specific and so will give accurate information on the prosthetic group of the protein, which can constitute an advantageous alternative to classical methods for detection of these hemoproteins.