Ecophysiological and phytochemical response to ozone of wine grape cultivars of Vitis vinifera L.

Vitis vinifera sensitivity to tropospheric ozone (O₃) has been evidenced in several studies. In this work, physiological and metabolic effects of O₃ on two wine cultivars of V. vinifera (i.e. Maturano and San Giuseppe) have been studied. Moreover, chlorogenic acid (CGA) production, in consideration of its importance in the biosynthetic pathway of polyphenols and as antioxidant, has been investigated. Maturano cultivar resulted more sensitive to O₃, as evidenced by the gas exchange reduction at the early stage of treatment, and by the increase in Ci/Ca and the decoupling of net photosynthesis and the stomatal conductance at the end of the treatment. Unexpectedly, O₃ did not activate stilbene production. Ozone induced an early CGA decrease, significantly more consistent in cv. Maturano, and an increase after 8 days, more consistent in cv. S. Giuseppe. These results suggest that CGA could be considered a biochemical marker of O₃-induced stress in V. vinifera.     

Unexpected one-step synthesis of 3-benzoyl-2-phenylbenzofurans under Wittig conditions

The reaction of 2-hydroxybenzyltriphenylphosphonium bromide with substituted benzoyl chlorides under Wittig conditions, led to 2-phenylbenzofuran derivatives 4a–p and the unexpected formation of 3-benzoyl-2-phenylbenzofuran derivatives 5a–p. Benzoyl chlorides possessing electron-withdrawing groups afforded 3-benzoyl-2-phenylbenzofuran derivatives in higher yields than those with electron-donating groups. This reaction represents a simple and regioselective, one-pot route towards the preparation of deactivated 3-benzoyl-2-phenylbenzofuran compounds which are difficult to obtain by the direct acylation of 2-phenylbenzofurans.     

Development of a CE-ESI-microTOF-MS method for a rapid identification of phenolic compounds in buckwheat

A sensitive CE-ESI-MS analytical method for the identification of buckwheat antioxidants has been developed. CE and ESI-TOF parameters (e.g. buffer composition and pH, sheath liquid composition, sheath liquid and gas flow rates, electrospray voltage) were optimized to obtain an optimal analytical separation and identification. The results confirmed the presence of phenolic acids, procyanidins and galloylated propelargonidins. The identification of swertiamacroside and 2-hydroxy-3-O-β-D-glucopyranosil-benzoic acid, found for the first time in our previous work, has been confirmed. Furthermore, 5,7,4'-trimethoxyflavan and dihydroxy-trimethoxyisoflavan have also been tentatively identified for the first time in buckwheat.     

Pressurized liquid extraction of lipids for the determination of oxysterols in egg-containing food

Pressurized liquid extraction (PLE, ASE) was compared with the Folch procedure (a solid-liquid extraction with chloroform/methanol 2:1, v/v) for the lipid extraction of egg-containing food; the accuracy of PLE for the quantitative determination of oxysterols in whole egg powder was evaluated. Samples of spray-dried whole egg, an Italian vanilla cake (Pandoro) and egg noodles were used. Two different extraction solvents (chloroform/methanol 2:1, v/v, and hexane/isopropanol 3:2, v/v) were tested at different extraction temperatures and pressures (60 degrees C at 15 MPa, 100 degrees C at 15 MPa, 120 degrees C at 20 MPa). No significant differences in the lipid recovery of the egg powder sample using PLE were found. However, PLE of the vanilla cake and egg noodles with the chloroform/methanol mixture was not selective enough and led to the extraction of a non-lipid fraction, including nitrogen-containing compounds. In the same samples, the pressurized hexane/isopropanol mixture gave a better recovery result, comparable to that obtained using the Folch method. Cholesterol oxidation products of the Folch extract and the pressurized liquid extract of spray dried egg powder (obtained with hexane/isopropanol 3:2, v/v, at 60 degrees C and 15 MPa) were determined by gas chromatography. PLE performed under these conditions is suitable to replace the Folch extraction, because the differences between the two methods tested were not statistically significant. Moreover, PLE shows important advantages, since the analysis time was shortened by a factor of 10, the solvent costs were reduced by 80% and the use of chlorinated solvents was avoided.     

Identification of plant sterols in hexaploid and tetraploid wheats using gas chromatography with mass spectrometry

Free sterols from hexaploid and tetraploid free-threshing wheats (Triticum aestivum L. and T. durum Desf.) and from their respective hulled wheats (T. spelta L. and T. dicoccon Schrank) were analysed by gas chromatography with mass spectrometry. The qualitative analysis of sterols showed a similar pattern either between hexaploid (T. aestivum, T. spelta) and tetraploid (T. durum, T. dicoccon) wheats or between free-threshing (T. aestivum, T. durum) and hulled (T. spelta, T. dicoccon) wheats. However, quantitative differences were found between tetraploid and hexaploid wheats, in that free sterol amounts in tetraploid wheats were 40% higher than in hexaploid ones. The mass spectra of the sterols were classified into four groups, taking into account the structural features of rings A and B. Typical mass spectral fragmentations of the four classes, and additional evidence related to the side chain of each molecule, were investigated together with their chromatographic behaviour, allowing identification of all the detected sterols.     

Harmonization of methods for analysis of cholesterol oxides in foods--the first portion of a long road toward standardization: interlaboratory study

A compilation of literature data on the content of cholesterol oxidation products (COP) in various food products and in blood demonstrates a large variation in content in products or tissues of very similar nature when analyzed in different laboratories according to a large number of methods. The lack of validated, internationally recognized methodology with published accuracy and precision has so far hindered such assessments. Hence an interlaboratory comparision of methodologies of COP analysis was undertaken on egg yolk powders (EYP), whole milk powders (WMP), skim milk powders (SMP), and lard (L). Each product type had one fresh sample (low) and one aged (high) in COP contents. A total of 17 sets of results on WMP, 15 on SMP and EYP, and 13 on L were compared. Overall results (mg/kg sample) varied extensively: Fresh EYP 0.72-265, aged EYP 2.51-361; fresh WMP 0.02-18.1, aged WMP 0.02-26.9; fresh SMP 0.02-6.51, aged SMP <0.01-6.51; fresh L 0.18-97, aged L 4.15-452. Some results were questioned, viz., those from laboratories not indicating substantial differences between samples "low" and "high" in total COP. Others were excluded because of lack of verification of identity of gas chromatographic peaks by mass spectrometry. Then a more narrow range of core results (mg/kg sample) was observed: Fresh EYP 5.69-29.5 sample, aged EYP 11.8-79.0; fresh WMP 0.12-1.76, aged WMP 1.17-13.7; fresh SMP <0.30-<1.21, aged SMP 0.30-2.26; fresh L 0.18-5.07, aged L 94.4-231. At a workshop discussing the results, numerous recommendations were made toward more reliable methodology for determination of COP in foods.     

Free and bound phenolic compounds in barley (Hordeum vulgare L.) flours. evaluation of the extraction capability of different solvent mixtures and pressurized liquid methods by micellar electrokinetic chromatography and spectrophotometry

The solution speciation of metals is a critical parameter controlling the bioavailability, solution-solid phase distribution and transport of metals in soils. The natural metal-complexing ligands that exist in soil solution include inorganic anions, inorganic colloids, organic humic substances, amino acids (notably phytosiderophores and bacterial siderophores) and low-molecular mass organic acids. The latter two groups are of particular significance in the soil surrounding plant roots (the rhizosphere). A number of analytical methodologies, encompassing computational, spectroscopic, physico-chemical and separation techniques, have been applied to the measurement of the solution speciation of metals in the environment. However, perhaps with the exception of the determination of the free metal cation, the majority of these techniques rarely provide species specific information. High-performance liquid chromatography (HPLC) coupled to a sensitive detection system, such as inductively coupled plasma mass spectrometry (ICP-MS) or electrospray ionisation mass spectrometry (ESI-MS), offers the possibility of separating and detecting metal-organic acid complexes at the very low concentrations normally found in the soil environment. This review, therefore, critically examines the literature reporting the HPLC separation of metal-organic acid complexes with reference to thermodynamic equilibrium and kinetic considerations. The limitations of HPLC techniques (and the use of thermodynamic equilibrium calculations to validate analytical results) are discussed and the metal complex characteristics necessary for chromatographic separation are described.     

High-performance liquid chromatography determination of phenyllactic acid in MRS broth

Phenyllactic acid (PLA) is an organic acid produced by some strains of lactic acid bacteria (LAB) and concentrations higher than 7.5 mg/ml inhibit growth of moulds and yeasts. Since PLA can be used to select LAB, a rapid, simple and cheap method for its determination is desirable. Typical methods for its analysis in broth are time-consuming, analytically complicated, and have poor recoveries. Herein we propose a simple and rapid method that does not require extraction, but only microfiltration of broth before injection in HPLC. The improved chromatographic conditions allow separation and quantification of PLA with a recovery of 98.7%. The method is highly reproducible with an intraday repeatability of the total peak area of 2.00%, while an interday repeatability of 2.69%.     

Analysis of glycerophospho‐ and sphingolipids by CE

Glycerophospholipids are amphiphilic molecules possessing polar head groups with a glycerol backbone and nonpolar variable long‐chain fatty acids. Numerous molecular species are found in a single class of glycerophospholipid, conferring to these lipids a high structural diversity. They are major components of biological membranes and participate in important activities involving cell signaling and substrate transport. Sphingolipids consist of long‐chain bases linked by an amide bond to a fatty acid and via the terminal hydroxyl group to complex carbohydrate or phosphorus moieties, constituting a complex family of compounds that also present an enormous structural variability. As important component of neuronal membranes, sphingolipids contribute to cellular diversity and functions and are associated with several neurodegenerative disorders. Moreover, they were studied in several foods due to their sensorial, reological, and antioxidant characteristics. In this work, the most relevant information available on glycerophospholipid and sphingolipid analysis by CE is reviewed. CE is a very promising analytical technique in polar lipid analysis, which provides high efficiency, relatively high resolution, and enormous versatility and requires small amounts of sample and solvent. MEKC and NACE methodologies have been developed as the most useful alternatives for these analyses by CE. Very interesting LODs have been achieved enabling the application of CE to the determination of glycerophospholipids and sphingolipids in several food and biological matrices.