Ana I. Castellote

Analysis of virgin olive oil volatile compounds by headspace solid-phase microextraction coupled to gas chromatography with mass spectrometric and flame ionization detection

The efficiency of headspace solid-phase microextraction (SPME) was evaluated for the qualitative and semi-quantitative analysis of virgin olive oil volatile compounds. The behaviour of four fibre coatings was compared for sensitivity, repeatability and linearity of response. A divinylbenzene-Carboxen-polydimethylsiloxane fibre coating was found to be the most suitable for the analysis of virgin olive oil volatiles. Sampling and chromatographic conditions were examined and the SPME method, coupled to GC with MS and flame ionization detection, was applied to virgin olive oil samples. More than 100 compounds were isolated and characterised. The presence of some of these compounds in virgin olive oil has not previously been reported. The main volatile compounds present in the oil samples were determined quantitatively.

The effects of harvest and extraction methods on the antioxidant content (phenolics, α-tocopherol, and β-carotene) in virgin olive oil

We studied the effects of harvesting and two processing systems (two-phase centrifugation and three-phase centrifugation) on olive oil quality. Oils extracted from high quality olives do not differ in free acidity, peroxide value and ultraviolet light absorption. Nor was the fatty acid composition affected. However, the antioxidant content of the oil was higher from green olives than from ripe olives. On the other hand, we determined that neither extraction method affects the presence of α-tocopherol and β-carotene, however, the phenolic content is higher in the two-phase method due to the addition of lukewarm water that is used to dilute the olive paste.

Rapid determination of vitamin E in vegetable oils by reversed-phase high-performance liquid chromatography

A quick and direct method for measuring tocopherols (alpha, beta+gamma and delta) in vegetable oils has been developed using RP-HPLC with UV detection. Previous extraction of tocopherols is not required. The oil is diluted in hexane and an aliquot is mixed with ethanol containing an internal standard (alpha-tocopherol acetate). The chromatographic system consists of an ODS-2 column with a methanol-water mobile phase. Tocopherols are detected at 292 nm in less than 5 min after injection. The method is precise (RSD=2.69%) and has a high mean recovery (98.14%).

Effect of ingestion of virgin olive oil on human low-density lipoprotein composition

Objective: To measure the incorporation of oleic acid and antioxidants (phenols and vitamin E) to low density lipoprotein (LDL) after acute and short-term ingestion of virgin olive oil. To study whether this incorporation contributes to an increase in LDL resistance to oxidation.Setting: Department of Food and Nutrition, University of Barcelona, Spain and Department of Lipids and Cardiovascular Epidemiology, IMIM, Barcelona, Spain.Subjects: Sixteen healthy volunteers aged 25–65 y.Design and interventions: To observe the change in the fatty acid profile, vitamin E, phenolic compounds and LDL oxidation-related variables after the postprandial phase and after daily ingestion of olive oil for one week.Results: Few changes were observed in the postprandial phase. However, after a week of olive oil consumption there was an increase in oleic acid (P=0.015), vitamin E (P=0.047), phenolics (P=0.021) and lag time (P=0.000), and a decrease in the maximum amount of dienes (P=0.045) and oxidation rate (P=0.05).Conclusion: After ingestion of virgin olive oil, an increase in antioxidants and oleic acid in LDL was observed as well as an improvement of LDL resistance to oxidation. Our results support the idea that daily ingestion of virgin olive oil could protect LDL from oxidation.Sponsorship: This study was supported by a research grant from Spain (ALI 97-1607-C02-02).

Elevated Circulating LDL Phenol Levels in Men Who Consumed Virgin Rather Than Refined Olive Oil Are Associated with Less Oxidation of Plasma LDL

In human LDL, the bioactivity of olive oil phenols is determined by the in vivo disposition of the biological metabolites of these compounds. Here, we examined how the ingestion of 2 similar olive oils affected the content of the metabolic forms of olive oil phenols in LDL in men. The oils differed in phenol concentrations as follows: high (629 mg/L) for virgin olive oil (VOO) and null (0 mg/L) for refined olive oil (ROO). The study population consisted of a subsample from the EUROLIVE study and a randomized controlled, crossover design was used. Intervention periods lasted 3 wk and were preceded by a 2-wk washout period. The levels of LDL hydroxytyrosol monosulfate and homovanillic acid sulfate, but not of tyrosol sulfate, increased after VOO ingestion (P < 0.05), whereas the concentrations of circulating oxidation markers, including oxidized LDL (oxLDL), conjugated dienes, and hydroxy fatty acids, decreased (P < 0.05). The levels of LDL phenols and oxidation markers were not affected by ROO consumption. The relative increase in the 3 LDL phenols was greater when men consumed VOO than when they consumed ROO (P < 0.05), as was the relative decrease in plasma oxLDL (P = 0.001) and hydroxy fatty acids (P < 0.001). Plasma oxLDL concentrations were negatively correlated with the LDL phenol levels (r = -0.296; P = 0.013). Phenols in LDL were not associated with other oxidation markers. In summary, the phenol concentration of olive oil modulates the phenolic metabolite content in LDL after sustained, daily consumption. The inverse relationship of these metabolites with the degree of LDL oxidation supports the in vivo antioxidant role of olive oil phenolics compounds.

Plasma fatty acid composition, estimated desaturase activities, and their relation with the metabolic syndrome in a population at high risk of cardiovascular disease

BACKGROUND & AIMS The metabolic syndrome (MetS) is a clustering of various metabolic abnormalities which is associated with increased risk of cardiovascular disease (CVD) and type 2 diabetes mellitus. Due to its increasing prevalence, it has become an important public health concern. Altered fatty acid (FA) composition and desaturase activities have been associated with several metabolic diseases, including MetS. The aim of the present study was to evaluate the relationship of the plasma FA profile and desaturase activities with the MetS in a Mediterranean population at high risk of CVD. METHODS Baseline data from 427 participants aged 55-80 years who took part in the interventional PREDIMED study were obtained. Individual FA was determined in plasma and desaturase activities were estimated from product/precursor ratios. Odds ratios (OR) and partial correlation coefficients were used to examine these relations with MetS and its components, respectively. RESULTS We found higher levels of C14:0, C16:0, C16:1n-7, estimated Δ(9)- or stearoyl-CoA desaturase (SCD), and estimated Δ(6) desaturase (D6D), and lower levels of C18:2n-6 in people with MetS compared to those without it. After adjustment for several confounders, only higher quartiles of C14:0, C16:0, C16:1n-7, and D6D were found to be associated with an increasing prevalence of MetS, while higher quartiles of C18:2n-6 were inversely associated with MetS. High proportions of C14:0, C16:0, C16:1n-7, C20:3n-6, SCD, and D6D, and decreased proportions of C18:2n-6 and estimated Δ(5)-desaturase (D5D) were associated with adverse profiles of several metabolic risk factors. Women showed more unhealthy FA pattern and lipid profiles than men, but only among those with MetS. CONCLUSION A FA composition and estimated desaturase activities consisting in high levels of SFA, SCD and D6D, and low levels of PUFA and D5D are associated with increased MetS probability and are characteristic of people presenting MetS, especially women. These findings support those observed in non-Mediterranean populations in which an altered FA profile and estimated desaturase activities are associated with MetS.

Simultaneous determination of α-tocopherol and β-carotene in olive oil by reversed-phase high-performance liquid chromatography

Abstract A reversed-phase high-performance liquid chromatographic method was developed for the determination, in one run, of α-tocopherol and β-carotene in virgin olive oil. The method involved a rapid saponification and a later extraction with a mixture of hexane–ethyl acetate. The chromatographic system consists of an ODS-2 column with a mobile phase of methanol–water–butanol and a diode-array detector. Linearity, precision, recovery and sensitivity were satisfactory. The main advantage of the proposed method is the speed and simultaneous determination of both compounds at the same time.

Changes in the phenolic content of low density lipoprotein after olive oil consumption in men. A randomized crossover controlled trial.

Olive oil decreases the risk of CVD. This effect may be due to the fatty acid profile of the oil, but it may also be due to its antioxidant content which differs depending on the type of olive oil. In this study, the concentrations of oleic acid and antioxidants (phenolic compounds and vitamin E) in plasma and LDL were compared after consumption of three similar olive oils, but with differences in their phenolic content. Thirty healthy volunteers participated in a placebo-controlled, double-blind, crossover, randomized supplementation trial. Virgin, common, and refined olive oils were administered during three periods of 3 weeks separated by a 2-week washout period. Participants were requested to ingest a daily dose of 25 ml raw olive oil, distributed over the three meals of the day, during intervention periods. All three olive oils caused an increase in plasma and LDL oleic acid (P < 0.05) content. Olive oils rich in phenolic compounds led to an increase in phenolic compounds in LDL (P < 0.005). The concentration of phenolic compounds in LDL was directly correlated with the phenolic concentration in the olive oils. The increase in the phenolic content of LDL could account for the increase of the resistance of LDL to oxidation, and the decrease of the in vivo oxidized LDL, observed in the frame of this trial. Our results support the hypothesis that a daily intake of virgin olive oil promotes protective LDL changes ahead of its oxidation.

Influence of variety and geographical origin on the lipid fraction of hazelnuts (Corylus avellana L.) from Spain: (III) oil stability, tocopherol content and some mineral contents (Mn, Fe, Cu)

Abstract Induction time, acid value and α-tocopherol content of hazelnut oil and some mineral contents (manganese, iron and copper) of hazelnut kernels cultivated in Catalonia (Spain) are determined. Analysis of variance (ANOVA) showed that statistically significant differences existed for: α-tocopherol content, acid value and copper content in relation to the harvesting year of samples. Significant differences were also found for: induction time, manganese and copper contents in relation to the location of samples. On the other hand, no significant differences were found between varieties. In addition, a correlation study was performed between all parameters included in this work. A strong negative correlation was observed between linoleic acid and manganese content, and also copper content and oil stability (Rancimat).

Triacylglycerol markers of mature human milk

Objectives: To determine which triacylglycerol (TAG) species in mature human milk are less affected by external factors and may thus be considered as TAG markers, as well as to determine which species are most influenced by these external conditions. Furthermore, we examine the correlation between the TAG markers and their fatty acids (FAs).Setting and subjects: Six healthy women from Barcelona (Catalonia, Spain).Design and interventions: In order to obtain the maximum variability of sampling conditions, 40 mature human milk samples were collected from different mothers, on different days, at different times of the day, and from different breasts during and after both the babys and mothers meal. TAG and FA profiles were determined and correlated. The TAG composition was determined by high-performance liquid chromatography with an evaporative light-scattering detector, and also with atmospheric pressure chemical ionisation mass spectrometry. FAs compositions were determined by gas chromatography.Results: The results were analysed using the SPSS statistical package and proved to be more variable than might have been found in a more restrictive sample design. Nevertheless, despite these conditions, some TAG species were found in relatively constant levels in mature human milk, and could thus be considered as markers of the mature milk TAG profile. TAG species that we can classify in this group were: LaMO, CaPO, LaCaO, LaPCa, LaOL, MPLn, LLO, LaOO, MPL, and MOL. The names do not indicate the location of fatty acids in the glycerol molecule. On the other hand, concentrations of other TAG species vary considerably between samples and consequently these may be understood to be especially affected by the external factors. TAGs like PaLS, MPO, PaOO, PPP, MPS, SPP, LOO, PPO, MOS, SSP, POL, and SOS are in this second group. Correlation between the TAG markers and their FAs was examined by Pearsons test and a significant correlation was found for some FAs.Conclusions: The TAG species present in mature human milk are affected in different ways by external factors such as dietary intake, nutritional status, length of lactation, time of the day, etc. Some TAGs may be considered as markers of mature human milk as they are relatively constant under a wide range of sampling conditions and do not depend on the factors mentioned.Sponsorship: This study was supported by the Fundació Mestres Jané.