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).

Rapid high-performance liquid chromatographic method for the simultaneous determination of retinol, alpha-tocopherol and beta-carotene in human plasma and low-density lipoproteins.

A reversed-phase HPLC method with diode-array detection was used to simultaneously determine retinol, alpha-tocopherol and beta-carotene in human plasma and low-density lipoproteins. An aliquot of sample was de-proteinized with ethanol containing beta-tocopherol acetate as internal standard, and the analytes were extracted twice with hexane. The solvent was evaporated to dryness under a stream of nitrogen and the residue was redissolved in methanol to be injected directly into the HPLC system. A multiple solvent system based on methanol, butanol and water at a flow-rate of 2 ml/min and held at 45 degrees C provided clear separation of these compounds in only 8 min. The method showed good linearity, precision and accuracy for all compounds. Owing to its simplicity, this method may be useful in routine clinical and epidemiological work.

Method for the quantitative extraction of resveratrol and piceid isomers in grape berry skins. Effect of powdery mildew on the stilbene content.

A simple method for the quantitative extraction of resveratrol and its glycosides from grape berry skins has been developed. Optimal conditions for the extraction were investigated. Type of solvent, time, and temperature assayed influenced resveratrol and piceid yield. Adequate extraction was attained with ethanol/water (80:20 v/v) maintained at 60 degrees C for 30 min. Recovery (>96%) and reproducibility (6.83-15.13%) were satisfactory. After extraction, resveratrol and piceid isomers were quantified by high-performance liquid chromatography coupled to a ultraviolet-visible diode array detector. The amounts detected in 13 samples of 7 different varieties analyzed were, on average, 92.33 microg/g of dry skin for cis-piceid, 42.19 microg/g of dry skin for trans-piceid, and 24.06 microg/g of dry skin for trans-resveratrol. cis-Resveratrol was not detected in any sample. In grape berries infected by powdery mildew the contentw of these compounds were considerably increased and the degree of infection was positively related to their stilbene content.

[Coronary disease protective factors: antioxidant effect of olive oil]. / Facteurs protecteurs de la maladie coronarienne: effet antioxydant de l'huile d'olive.

Alongside the French paradox, the REGICOR Study (Girona, Spain) has shown another paradox in the Mediterranean area: a high prevalence of cardiovascular risk factors with low incidence of myocardial infarction in the population of Girona, Spain. The antioxidant effects associated with olive oil consumption could explain part of this 'Mediterranean Paradox'. Virgin olive oils processed by two centrifugation phases and with low fruit ripeness have the highest levels of antioxidant content. The total content of phenolic compounds (PC) from virgin olive oil could delay LDL oxidation. The content and nature of olive oil PC have a high influence in the antioxidant capacity of an olive oil. PC from diet could bind human LDL in non-supplemented volunteers. PC from virgin olive oil could bind LDL and tyrosol is bioavailable in humans from ingestion of virgin olive oil in its natural form.

Resveratrol and piceid levels in natural and blended peanut butters.

Peanut and its derivatives, especially peanut butter, are extensively consumed in many countries, mainly in the United States, which is also the major exporter of these products. trans-Resveratrol is present in peanuts, and recently this compound has been quantified in peanut butter. It is well-known that there are beneficial effects of trans-resveratrol and its glucoside, the piceid, in health. The absorption of trans-resveratrol has been proven in animals, and certain studies show that the absorption of some phenols is enhanced by conjugation with glucose, so that it could be possible that trans-piceid would be more absorbed than its aglycon (trans-resveratrol). In our work, we have identified the presence of trans-piceid in peanut butter with a new method to quantify trans-resveratrol and trans-piceid (3-beta-glucose of trans-resveratrol). This fact is very interesting because the glucosilated form could be more easily absorbed by the intestinal gut; in this way trans-piceid would exercise its beneficial function more efficiently than trans-resveratrol. To our knowledge, this is the first time that trans-piceid has been quantified in peanut butter. Resveratrol and piceid contents in natural peanut butters were found to be significantly higher than those in blended peanut butters.

Piceid, the major resveratrol derivative in grape juices.

The levels of trans-piceid, cis-piceid, trans-resveratrol, and cis-resveratrol have been measured in 36 grape juices using an HPLC system with spectral analysis of eluting peaks. The piceid (glucosides) were the major component in the grape juices. In red grape juices the average concentrations were 3.38 mg/L for trans-piceid, 0.79 mg/L for cis-piceid, 0.50 mg/L for trans-resveratrol, and 0.06 mg/L for cis-resveratrol. In white grape juices the levels were, on average, 0.18 mg/L for trans-piceid, 0.26 mg/L for cis-piceid, and 0.05 mg/L for trans-resveratrol, and cis-resveratrol was not detected in any sample. Levels of total resveratrol (trans- and cis-resveratrol and -piceid) found in red and in white grape juices are similar to those described in Spanish red and white wines. Due to their resveratrol content, as well as other phenolics, grape juices may have a beneficial health effect of interest to those who cannot drink wine.

Beneficial effects of white wines.

Wine phenolics have been reported to have health benefits, including protection against cardiovascular diseases and anticarcinogenic effects. White wines are usually made with the free-running juices without pomace, which has no contact with the grape skins. This is the main reason that the phenolic content of white wines is lower than that of red wines. However, white wine phenols have a comparable or higher antioxidant capacity than red wine phenols. Therefore, it is important to determine which phenolic compounds are present in white wines and which factors affect phenolic composition. We studied the influence of several factors, including variety and different technological processes, on phenolic composition. Significant differences were observed when any of these variables were considered. Consequently, if wine makers were to take these factors into consideration, the possible beneficial effects of their wines could be increased.

Scientific basis for the health benefits of the Mediterranean diet.

The meaning and influence of the recently introduced concept of the "Mediterranean diet" are explained herein. Although now viewed as a discovery, the Mediterranean people's surprise at this "new" concept is stressed as they are already well acquainted with the qualities of this dietary model. Important changes in lifestyle and nutrition are currently taking place, which may not be the best way to maintain a healthy diet. Possibly, too many ancient nutritional traditions have been abandoned. If this is so, the return to this old dietary model, which has proven to be so beneficial to health, should be welcomed. To understand the significance of the Mediterranean diet the most beneficial compounds and details of the Mediterranean "classical model of lifestyle and nutrition" are explained. Finally, hypotheses about the scientific basis of the benefits of the Mediterranean diet are explained.

Determination of trans-resveratrol in plasma by HPLC.

trans-Resveratrol (trans-3,5,4'-trihydroxystilbene), a phenolic compound present in grapes, wines, and peanuts, has been reported to have health benefits including anticarcinogenic effects and protection against cardiovascular diseases. Despite its importance, little is known about its bioavailability in both humans and animals. A fundamental step for this evaluation consisted in measuring this stilbene in blood. In the present work, a simple and rapid HPLC method with diode array-UV detection has been developed. Resveratrol contained in plasma was purified by solid-phase extraction using a C18 cartridge. The sample was rinsed with water and methanol-water (25:75 v/v), and trans-resveratrol was finally eluted with methanol. The collected fraction was evaporated under nitrogen and analyzed by HPLC. The method was validated by obtaining a linear correlation, a detection limit of 20 micrograms/L, and a good precision with a coefficient of variation of 2.85%. trans-Resveratrol administered orally to rats was detected in plasma. With this procedure, excellent separation of trans-resveratrol is achieved, thus allowing a rapid analysis of the sample for absorption, distribution, and metabolism studies.

Comparison of two methods for the determination of fatty acid profiles in plasma and erythrocytes.

We have validated and compared two methods for the determination of fatty acid profiles in biological samples by capillary gas chromatography. Method I consisted of a previous lipid extraction and esterification of fatty acids using boron trifluoride-methanol. Method II was a direct method that combined extraction and esterification of freeze-dried samples in a single step, using acetyl chloride as the reagent. The two methods were applied to the analysis of plasma and erythrocyte samples. Both methods gave similar results in plasma, whereas in erythrocytes, the direct method gave significantly higher contents of total fatty acids. Precision and recovery rates were determined and the results were satisfactory. Detection and quantification limits showed that both methods had excellent sensitivity. It was concluded that the direct method has substantial advantages over the conventional method, such as higher values in erythrocytes, rapidity and less possibility of contamination or fatty acid losses. Therefore, it is preferable for the analysis of biological samples such as plasma and erythrocytes.

Comparison of two methods for the determination of fatty acid profiles in plasma and erythrocytes.

We have validated and compared two methods for the determination of fatty acid profiles in biological samples by capillary gas chromatography. Method I consisted of a previous lipid extraction and esterification of fatty acids using boron trifluoride-methanol. Method II was a direct method that combined extraction and esterification of freeze-dried samples in a single step, using acetyl chloride as the reagent. The two methods were applied to the analysis of plasma and erythrocyte samples. Both methods gave similar results in plasma, whereas in erythrocytes, the direct method gave significantly higher contents of total fatty acids. Precision and recovery rates were determined and the results were satisfactory. Detection and quantification limits showed that both methods had excellent sensitivity. It was concluded that the direct method has substantial advantages over the conventional method, such as higher values in erythrocytes, rapidity and less possibility of contamination or fatty acid losses. Therefore, it is preferable for the analysis of biological samples such as plasma and erythrocytes.

Administration of low doses of fish oil derived N-3 fatty acids to elderly subjects.

OBJECTIVES: To assess the incorporation of n-3 polyunsaturated fatty acids (PUFA) in plasma and erythrocyte lipids of elderly subjects after ingestion of very low doses of fish oil. The effects on alpha-tocopherol and retinol concentrations were also studied. SETTING: Municipal nursing home in Barcelona, Spain. SUBJECTS: Forty-five elderly subjects aged 60-92 y. DESIGN AND INTERVENTION: Subjects received a non-commercialized milk formula containing 1% fish oil for 15 months, which provided 0.40 g/d of n-3 PUFA. Fatty acid profiles and antioxidant concentrations were measured before and after the intervention period. RESULTS: Fish oil ingestion was associated with significant increases in total n-3 PUFA in plasma and erythrocytes by 32% and 18%, respectively. Eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid concentrations were higher after the ingestion period both in plasma and erythrocytes (P < 0.05), whereas linoleic and arachidonic acids remained unchanged. The n-6/n-3 ratio decreased by 21% in plasma and by 16% in erythrocytes (P < 0.05). Moreover, younger subjects showed a greater incorporation of EPA and DHA than older subjects. Plasma alpha-tocopherol and retinol concentrations did not vary significantly, whereas erythrocyte alpha-tocopherol was significantly higher after the intervention period. CONCLUSION: This study shows that low doses of n-3 PUFA supplemented with adequate amounts of alpha-tocopherol can be incorporated into blood lipids in elderly subjects without lowering their antioxidant concentrations.

Determination of sterol content in different food samples by capillary gas chromatography.

An accurate method for the determination of sterols by capillary gas chromatography was developed and applied to the analysis of food. The procedure includes the following steps: dichloromethane-methanol (2:1, v/v) lipid extraction, saponification at 80 degrees C and separation of the unsaponifiable matter with cyclohexane, derivatization to form trimethylsilyl ethers and gas chromatography using 5 alpha-cholestane as the internal standard. The method shows good accuracy, precision and sensitivity and is suitable for the determination of sterols in food.