Determination of polyphosphates in products of animal origin: application of a validated ion chromatography method for commercial samples analyses

A reliable and accurate analytical method was developed and validated for the quantitative determination of polyphosphates (diphosphate, triphosphate, trimetaphosphate and tetrapolyphosphate) in products of animal origin (meat, dairy and fish products) by ion chromatography with suppressed conductivity detection. The chromatographic separations were accomplished by using an anion-exchange column eluted with a sodium hydroxide gradient. The method validation, performed according to Regulation 882/2004/EC and Decision 657/2002/EC, provided results conform with the European Directives with respect to linearity (R?>?0.996), specificity, precision (CV????4.5?%), recovery (ranging from 87.2 to 101.1?%), detection and quantification limits and ruggedness. The method reliability was confirmed evaluating the method measurement uncertainty, lower than 7.5?% and by proficiency test results. Finally, the method ability to discriminate samples treated from those not-treated with polyphosphates was verified by analyzing commercial samples containing polyphosphates (cooked ham, wurstel, corned beef, processed cheese and surimi) or treated in-house with polyphosphates (pangasius fillets, shrimps and cuttlefishes).     

Morphology and surface properties of some siloxane–organic copolymers

BACKGROUND: It is well known that, due to their extremely low polarity, polysiloxanes are incompatible with almost any organic system. This incompatibility leads to phase separation in mixed siloxane–organic systems. RESULTS: Three siloxane–organic copolymers, poly[(5,5′‐methylene‐bis‐salicylaldehyde)‐imine‐(1,3‐bis(propylene)tetramethyldisiloxane)] (Paz1), poly[(2,5‐dihydroxy‐1,4‐benzoquinone)‐imine‐(1,3‐bis(propylene)tetramethyldisiloxane)] (Paz2) and poly[1,3‐bis(propylene)tetramethyldisiloxane diamide] (Pam), were prepared by the reaction of 1,3‐bis(3‐aminopropyl)tetramethyldisiloxane with appropriate organic partners (5,5′‐methylene‐bis‐salicylaldehyde, 2,5‐dihydroxy‐1,4‐benzoquinone and oxalyl chloride, respectively). The morphologies dictated by the incompatibility between siloxane and polar organic moieties were investigated using differential scanning calorimetry and scanning electron microscopy. The surface activity of the copolymers and water vapour sorption capacity were also measured. CONCLUSION: Even though the polar sequences are very short ones, the highly flexible siloxane‐containing sequence permits the self‐assembly of these into more or less polar domains. Such an organization influences the properties of the resulting materials, an important place being occupied by the surface properties. Copyright © 2009 Society of Chemical Industry     

Unconventional method used in synthesis of polyphosphoesters

Polyphosphonates, polyphosphates, polyarylazophosphonates and polyarylazophosphates were synthesized by a new alternative to the classical phase transfer catalysis, respectively, the modified inverse phase transfer catalysis (IPTC) polycondensation of various phenylphosphonic (phosphoric) dichlorides (phenylphosphonic dichloride, phenylphosphoric dichloride, p-chlorophenylphosphoric dichloride) with bisphenols: bisphenol A, tetrabromobisphenol A, bis-(4-hydroxyphenyl)methane (bisphenol F), 4,4′-dihydroxyazobenzene. The polymers were characterized by infrared spectroscopy and magnetic resonance (¹H-NMR, ³¹P-NMR, ¹³C-NMR) spectroscopy. Yields in the range of 63.5–85% and molecular weights (M _w) of ~2,000–8,200 g mol^?1 were obtained. Polyphosphonates were stable up 210–270 °C and polyphosphates up 190–220 °C in air atmosphere. For a correct estimation of the thermal behavior of similar compounds, a kinetic analysis using a modified version of Non Parametric Kinetic method for representative polyphosphonate and polyphosphate was realized. The studies made on the hydrolytic degradation of the synthesized polyphosphates show that the most stable polymer under alkali-catalyzed degradation is the polyphosphate obtained by IPTC of phenylphosphoric dichloride and bisphenol A.     

Solubility Limits of Cholesterol, Lanosterol, Ergosterol, Stigmasterol, and β-Sitosterol in Electroformed Lipid Vesicles

Here we use nuclear magnetic resonance to measure the solubility limit of several biologically relevant sterols in electroformed giant unilamellar vesicle membranes containing phosphatidylcholine (PC) lipids in ratios of 1:1:X DOPC:DPPC:sterol. We find solubility limits of cholesterol, lanosterol, ergosterol, stigmasterol, and β-sitosterol to be 65-70%, ~35%, 30-35%, 20-25%, and ~40%, respectively. The low solubilities of stigmasterol and β-sitosterol, which differ from cholesterol only in their alkyl tails, show that subtle differences in tail structure can strongly affect sterol solubility. Below the solubility limits, the fraction of sterol to PC-lipid in electroformed vesicles linearly reflects the fraction in the original stock solutions used in the electroformation process.     

Harmful dinoflagellate Ostreopsis cf. ovata Fukuyo: detection of ovatoxins in field samples and cell immunolocalization using antipalytoxin antibodies

Ostreopsis cf. ovata, a benthic dinoflagellate often blooming along the Mediterranean coasts, has been associated with toxic events ranging from dyspnea to mild dermatitis. In late September 2009, an Ostreopsis cf. ovata bloom occurred in the Gulf of Trieste (Northern Adriatic Sea; Italy), causing pruritus and mild dermatitis in beachgoers. An integrated study was initiated to characterize Ostreopsis cells by light and confocal microscopy, PCR techniques, immunocytochemistry, and high resolution liquid chromatography-mass spectrometry (HR LC-MS). The presence of Ostreopsis cf. ovata of the Atlantic/Mediterranean clade was unambiguously established by morphological and genetic analyses in field samples. Several palytoxin-like compounds (ovatoxin-a,-b,-c,-d,-e) were identified by HR LC-MS, ovatoxin-a being the most abundant (45-64 pg/cell). Surprisingly, no palytoxin was detected. For the first time, monoclonal and polyclonal antipalytoxin antibodies revealed the intracellular cytoplasmic localization of ovatoxins, suggesting their cross-reactivity with these antibodies. Since harmful dinoflagellates do not always produce toxins, the immunocytochemical localization of ovatoxins, although qualitative, can provide an early warning for toxic Ostreopsis cells before their massive diffusion and/or concentration in seafood.     

Immunocytochemical localization of the NMDA-R2A receptor subunit in the cat retina

Immunocytochemical studies were performed to determine the distribution and cellular localization of the NMDA-R2A receptor subunit (R2A) in the cat retina. R2A-immunoreactivity (R2A-IR) was noted in all layers of the retina, with specific localizations in the outer segments of red/green and blue cone photoreceptors, B-type horizontal cells, several types of amacrine cells, Müller cells and the majority of cells in the ganglion cell layer. In the inner nuclear layer, 48% of all cells residing in the amacrine cell layer were R2A-IR including a cell resembling the GABAergic A17 amacrine cell. Interestingly, the AII rod amacrine cell was devoid of R2A-IR. Although the localization of the R2A subunit was anticipated in ganglion cells, amacrines and Müller cells, the presence of this receptor subunit to the cells in the outer retina was not expected. Here, both the R2A and the R2B subunits were found to be present in the outer segments of cone photoreceptors and to the tips of rod outer segments. Although the function of these receptor subunits in rod and cone photoreceptors remains to be determined, the fact that both R2A and R2B receptor subunits are localized to cone outer segments suggests a possible alternative pathway for calcium entry into a region where this cation plays such a crucial role in the process of phototransduction. To further classify the cells that display NR2A-IR, we performed dual labeling experiments showing the relationship between R2A-labeled cells with GABA. Results showed that all GABAergic-amacrines and displaced amacrines express the R2A-subunit protein. In addition, approximately 11% of the NR2A-labeled amacrines, did not stain for GABA. These findings support pharmacological data showing that NMDA directly facilitates GABA release in retina and retinal cultures [I.L. Ferreira, C.B. Duarte, P.F. Santos, C.M. Carvalho, A.P. Carvalho, Release of [3H]GABA evoked by glutamate receptor agonist in cultured chick retinal cells: effect of Ca2+, Brain Res. 664 (1994) 252-256; G.D. Zeevalk, W.J. Nicklas, Action of the anti-ischemic agent ifenprodil on N-methyl-d-aspartate and kainate-mediated excitotoxicity, Brain Res. 522 (1990) 135-139; R. Huba, H.D. Hofmann, Transmitter-gated currents of GABAergic amacrine-like cells in chick retinal cultures, Vis. Neurosci. 6 (1991) 303-314; M. Yamashita, R. Huba, H.D. Hofmann, Early in vitro development of voltage- and transmitter-gated currents in GABAergic amacrine cells, Dev. Brain Res. 82 (1994) 95-102; R. Ientile, S. Pedale, V. Picciurro, V. Macaione, C. Fabiano, S. Macaione, Nitric oxide mediates NMDA-evoked [3H]GABA release from chick retina cells, FEBS Lett. 417 (1997) 345-348; R.C. Kubrusly, M.C. deMello, F.G. deMello, Aspartate as a selective NMDA agonist in cultured cells from the avian retina, Neurochem. Intl. 32 (1998) 47-52] or reduction of GABA in vivo [N.N. Osborn, A.J. Herrera, The effect of experimental ischaemia and excitatory amino acid agonist on the GABA and serotonin immunoreactivities in the rabbit retina, Neurosci. 59 (1994) 1071-1081]. Since the majority of GABAergic synapses in the inner retina are onto both rod and cone bipolar axon terminals [R.G. Pourcho, M.T. Owzcarzak, Distribution of GABA immunoreactivity in the cat retina: A light and electron-microscopic study, Vis. Neurosci. 2 (1989) 425-435], we hypothesize that the NMDA-receptor plays a crucial role in providing feedback inhibition onto rod and cone bipolar cells.     

Behavior of fat constitutents during refining. 8. Determination of polymeric fatty acids in rape seed oil and their behavior during refining]

1. Studies on rape-seed oil have shown that the determination of oxypolymers by means of the method according to Rost (determination of fatty acids insoluble in petroleum ether) yields unsatisfactory results. By way of contrast, the gravimetric determination (after isolation of the polymers via the non-urea adduct fatty acid methyl esters and thin-layer chromatographic separation), as proposed by the authors, permits to obtain results of good reproducibility and to determine thermopolymers and oxypolymers separately. 2. During the refining of rape-seed oil, it was observed that its polymer content decreased considerably on bleaching, and increased on deodorizing.