Raffaele Sacchi

1H and 13C NMR of virgin olive oil. An overview

The authentication and quality assessment of virgin olive oil were performed using high‐resolution 1H and 13C NMR spectroscopy. An overview of the various determinations currently assessed is presented with emphasis on the detection of adulteration with foreign (seed) oils and esterified or refined olive and olive‐pomace oils. Recent results on the NMR analysis of natural compounds (diacylglycerols, free fatty acids, aldehydes, polyphenols, etc.) related to the quality‐freshness of virgin olive oil are also reported. The possible contribution of high‐resolution NMR to the authentication of geographical origin of virgin olive oil is discussed.

Characterization of the 12% trichloroacetic acid-insoluble oligopeptides of Parmigiano-Reggiano cheese

The isolation and identification of low molecular mass peptides formed during the ripening of Parmigiano-Reggiano cheese is described. A strategy was used based on the fractionation of nitrogenous material using chemical methods followed by HPLC to isolate peptides and fast atom bombardment-mass spectrometry to identify them. It was found that the majority of cheese oligopeptides arose from the proteolysis of β-casein. Several phosphopeptides and oligopeptides known in vivo to be biologically active have also been identified during the ripening of cheese.

A high-field1H nuclear magnetic resonance study of the minor components in virgin olive oils

High-field (600 MHz) nuclear magnetic resonance (NMR) spectroscopy was applied to the direct analysis of virgin olive oil. Minor components were studied to assess oil quality and genuineness. Unsaturated and saturated aldehyde resonances, as well as those related to other volatile compounds, were identified in the low-field region of the spectrum by two-dimensional techniques. Unsaturated aldehydes can be related to the sensory quality of oils. Other unidentified peaks are due to volatile components, because they disappear after nitrogen fluxing. The statistical analysis performed on the intensity of these peaks in several oil samples, obtained from different olive varieties, allows clustering and identification of oils arising from the same olive variety. Diacylglycerols, linolenic acid, other volatile components, water, acetic acid, phenols, and sterols can be detected simulteneously, suggesting a useful application of high-field NMR in the authentication and quality assessment of virgin olive oil.

Hydroxytyrosol, a phenolic compound from virgin olive oil, prevents macrophage activation

We investigated the effect of hydroxytyrosol (HT), a phenolic compound from virgin olive oil, on inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in J774 murine macrophages stimulated with lipopolysaccharide (LPS). Incubation of cells with LPS caused an increase in iNOS and COX-2 mRNA and protein level as well as ROS generation, which was prevented by HT. In addition, HT blocked the activation of nuclear factor-κB (NF-κB), signal transducer and activator of transcription-1α (STAT-1α) and interferon regulatory factor-1 (IRF-1). These results, showing that HT down-regulates iNOS and COX-2 gene expression by preventing NF-κB, STAT-1α and IRF-1 activation mediated through LPS-induced ROS generation, suggest that it may represent a non-toxic agent for the control of pro-inflammatory genes.

Characterization of the oligopeptides of Parmigiano-Reggiano cheese soluble in 120 g trichloroacetic acid/1

The non-protein nitrogen (NPN) of samples of Parmigiano-Reggiano cheese ripened for 6 and 15 months was fractionated by ion-exchange chromatography on a Cu(2+)-Chelex column to separate oligopeptides from free amino acids. Peptide components were isolated by reversed-phase HPLC and identified by fast atom bombardment-mass spectrometry (FAB-MS). Only the NPN fraction of 6 month old cheese samples contained enough peptides to be further characterized. On the basis of FAB-MNS spectral results, 39 oligopeptides were identified, the main components being phosphopeptides. Two sets of both intact and partly dephosphorylated peptides, accounting for a total of 19 phosphopeptides, were formed by the hydrolysis of beta-casein and belonged to regions 1-20 and 6-28 of beta-casein. The formation and potential role of these peptides in cheese is discussed.

Proton nuclear magnetic resonance rapid and structure-specific determination ofω-3 polyunsaturated fatty acids in fish lipids

Based on proton nuclear magnetic resonance (1H-NMR) spectroscopy, a rapid and structure-specific method for the determination of ω-3 polyunsaturated fatty acids (PUFAs) in fish lipids is presented. The different chemical shift observed for the methyl resonance of ω-3 PUFAs (δ=0.95 ppm) with respect to the methyl resonance of all other fatty acids (δ=0.86 ppm) has provided the possibility of proposing a new and rapid method for the determination of ω-3 PUFA content. Twenty-four fish lipid samples (raw, cooked and canned albacore tuna) produced results that showed good agreement between1H-NMR analysis and gas chromatographic determination. Raw and cooked samples showed significantly higher levels of ω-3 PUFA than canned tuna.

Relationship between Virgin Olive Oil Phenolic Compounds and Acrylamide Formation in Fried Crisps

In this paper the relationship between virgin olive oil (VOO) phenol compounds and the formation of acrylamide in potato crisps was investigated. The phenol compositions of 20 VOO samples were screened by LC-MS, and 4 oils, characterized by different phenol compound patterns, were selected for frying experiments. Slices of potatoes were fried at 180 degrees C for 5, 10, and 15 min, and acrylamide content was determined by LC-MS. Results demonstrated that VOO phenolic compounds are not degraded during frying, and crisp color was not significantly different among the four VOOs. Acrylamide concentration in crisps increased during frying time, but the formation was faster in the oil having the lowest concentration of phenolic compounds. Moreover, the VOO having the highest concentration of ortho-diphenolic compounds is able to efficiently inhibit acrylamide formation in crisps from mild to moderate frying conditions. It was concluded that the use of ortho-diphenolic-rich VOOs can be proposed as a reliable mitigation strategy to reduce acrylamide formation in domestic deep-frying.

Maldi-tof mass spectrometry profiling of polar and nonpolar fractions in heated vegetable oils.

Triacylglycerol oxidation of thermally stressed (6 h at 180 degrees C, simulating deep-frying conditions) edible vegetable oil (sunflower and olive) was studied using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Chromatographic separation of the nonpolar and polar components from the heated oil performed on silica gel prior to MS analysis significantly enhanced the detection of oxidized components. The spectra contained signals that were assigned to triacylglycerols (TAG), diacylglycerols (DAG), triacylglycerol oxidative dimers, oxidized TAG, and TAG fragments arising from the homolytic beta-scission of linoleyl, peroxy, and alkoxy radicals. Enrichment of the polar compounds prevented mass spectrometric ion suppression, thus allowing the detection of minor species originating from thermal oxidation. In addition, this allowed the monitoring of polar compounds in vegetable oils undergoing mild thermal treatment. As such, chromatographic separation coupled with MALDI-TOF MS analysis provided a rapid, sensitive, and specific tool to assess the thermal oxidation of vegetable oils.

Antioxidant activity of virgin olive oil phenolic compounds in a micellar system

The antioxidant activity of two virgin olive oils, obtained from the same olive (Olea europaea sativa) batch but processed with different hammer crushing conditions, was evaluated by measuring their protective action towards linoleic acid peroxidation in a micellar system. The antioxidant efficiency (AE) of the oil phenolic fraction was higher when the olives were processed with a higher hammer crusher rotation rate. HPLC analysis demonstrated that one of the main derivatives of oleuropein, indicated as 3,4-DHPEA-EDA (the dialdehydic form of elenolic acid linked with 3,4- dihydroxyphenylethanol), is present only in the oil with higher AE. 3,4-DHPEA-EDA showed the greatest antioxidant ability among virgin olive oil phenols. Its greater efficiency in the micellar system in comparison with 3,4-dihydroxyphenylethanol (3,4-DHPEA) is related to its greater lipophilicity. It is suggested that the behaviour in the water-micellar environment is representative of that in a real system such as tomato-based sauce with added virgin olive oil. # 1999 Society of Chemical Industry

Influence of some environmental factors on drupe maturation and olive oil composition

BACKGROUND Understanding the relationships between olive cultivars and the cultivation environment as well as optimising cultivation choices can lead to maximum expression of oil production in terms of both quantity and quality. For this purpose, samples of the Ortice olive cultivar grown in two different environments in southern Italy at altitudes of 500 and 50 m above sea level (a.s.l.) were harvested on various dates to monitor drupe maturation and determine the nutritional and chemical characteristics of the oils. RESULTS Fruits grown at 50 m a.s.l. ripened about 10-15 days earlier than those grown at 500 m a.s.l. The oil obtained at 500 m a.s.l. was characterised by a higher content of total polyphenols and a higher content of oleic and stearic acids. CONCLUSION The different heat accumulation related to the environment affected drupe development as well as the colouring trend and oil content. Consequently, the growth environment changed the content of Ortice oil fatty acids and polyphenols, while the flavour profile remained fairly stable in both environments, with the cultivar effect prevailing over the environment factor.