Alegría Carrasco-Pancorbo

Phenolic Molecules in Virgin Olive Oils: a Survey of Their Sensory Properties, Health Effects, Antioxidant Activity and Analytical Methods. An Overview of the Last Decade Alessandra

Among vegetable oils, virgin olive oil (VOO) has nutritional and sensory characteristics that to make it unique and a basic component of the Mediterranean diet. The importance of VOO is mainly attributed both to its high content of oleic acid a balanced contribution quantity of polyunsaturated fatty acids and its richness in phenolic compounds, which act as natural antioxidants and may contribute to the prevention of several human diseases. The polar phenolic compounds of VOO belong to different classes: phenolic acids, phenyl ethyl alcohols, hydroxy-isochromans, flavonoids, lignans and secoiridoids. This latter family of compounds is characteristic of Oleaceae plants and secoiridoids are the main compounds of the phenolic fraction. Many agronomical and technological factors can affect the presence of phenols in VOO. Its shelf life is higher than other vegetable oils, mainly due to the presence of phenolic molecules having a catechol group, such as hydroxytyrosol and its secoiridoid derivatives. Several assays have been used to establish the antioxidant activity of these isolated phenolic compounds. Typical sensory gustative properties of VOO, such as bitterness and pungency, have been attributed to secoiridoid molecules. Considering the importance of the phenolic fraction of VOO, high performance analytical methods have been developed to characterize its complex phenolic pattern. The aim of this review is to realize a survey on phenolic compounds of virgin olive oils bearing in mind their chemical-analytical, healthy and sensory aspects. In particular, starting from the basic studies, the results of researches developed in the last ten years will be focused.

High Capacity Capillary Electrophoresis-Electrospray Ionization Mass Spectrometry: Coupling a Porous Sheathless Interface with Transient-Isotachophoresis

A sheathless interface making use of a porous tip has been used for coupling capillary electrophoresis and electrospray ionization mass spectrometry. First, effective flow rates using the interface have been characterized. It was found that the interface is capable of generating a stable spray with flow rates ranging from below 10 nL/min to >340 nL/min, enabling its use in either the mass or concentration-sensitive region of the electrospray process. Subsequently, by analyzing peptide mixtures of increasing complexity, we have demonstrated that this platform provides exquisite sensitivity enabling the detection of very low amounts of materials with very high resolving power. Transient isotachophoresis (t-ITP) can also be integrated with this setup to increase the mass loading of the system while maintaining peak efficiency and resolution. Concentration limits of detection in the subnanomolar or nanomolar range can be achieved with or without t-ITP, respectively. The application of a vacuum at the inlet of the separation capillary further allowed the peak capacity of the system to be improved while also enhancing its efficiency. As a final step in this study, it was demonstrated that the intrinsic properties of the interface allows the use of coated noncharged surfaces so that very high peak capacities can be achieved.

Characterization and quantification of phenolic compounds of extra-virgin olive oils with anticancer properties by a rapid and resolutive LC-ESI-TOF MS method.

The characterization and quantification of extra-virgin olive oil (EVOO) phenolic compounds by a rapid resolution liquid chromatography (RRLC) method coupled to diode-array and time of flight mass spectrometry (TOF) detection systems was developed. The RRLC method transferred from a conventional HPLC one achieved better performance with shorter analysis times. The phenolic compounds were separated with a C18 column (150 mm x 4.6mm, 1.8 microm) using water with 0.5% acetic acid and acetonitrile as mobile phases. Good peak resolution was obtained and 19 different phenols were identified in less than 20 min providing a new level of information about the samples in shorter time. The applicability of this analytical approach was confirmed by the successful analysis of three different EVOO varieties (Picual, Hojiblanca, and Arbequina) obtained from different trademarks. Besides identification of the most important phenolic compounds and their quantification in three different ways (RRLC-UV, RRLC-MS and a new approach using the total polyphenol content obtained with Folin Ciocalteau, the relative areas and the response factors), we also described the occurrence of correlations between the phenolic composition of EVOO-derived crude phenolic extracts and their anti-proliferative abilities toward human breast cancer-derived cell lines. When compared with lignans-rich EVOO varieties, secoiridoids-rich EVOO had a significantly strong ability to alter cell viability in four different types of human breast carcinoma cells.

tabAnti-HER2 (erbB-2) oncogene effects of phenolic compounds directly isolated from commercial Extra-Virgin Olive Oil (EVOO)

BackgroundThe effects of the olive oil-rich Mediterranean diet on breast cancer risk might be underestimated when HER2 (ERBB2) oncogene-positive and HER2-negative breast carcinomas are considered together. We here investigated the anti-HER2 effects of phenolic fractions directly extracted from Extra Virgin Olive Oil (EVOO) in cultured human breast cancer cell lines.MethodsSolid phase extraction followed by semi-preparative high-performance liquid chromatography (HPLC) was used to isolate phenolic fractions from commercial EVOO. Analytical capillary electrophoresis coupled to mass spectrometry was performed to check for the composition and to confirm the identity of the isolated fractions. EVOO polyphenolic fractions were tested on their tumoricidal ability against HER2-negative and HER2-positive breast cancer in vitro models using MTT, crystal violet staining, and Cell Death ELISA assays. The effects of EVOO polyphenolic fractions on the expression and activation status of HER2 oncoprotein were evaluated using HER2-specific ELISAs and immunoblotting procedures, respectively.ResultsAmong the fractions mainly containing the single phenols hydroxytyrosol and tyrosol, the polyphenol acid elenolic acid, the lignans (+)-pinoresinol and 1-(+)-acetoxypinoresinol, and the secoiridoids deacetoxy oleuropein aglycone, ligstroside aglycone, and oleuropein aglycone, all the major EVOO polyphenols (i.e. secoiridoids and lignans) were found to induce strong tumoricidal effects within a micromolar range by selectively triggering high levels of apoptotic cell death in HER2-overexpressors. Small interfering RNA-induced depletion of HER2 protein and lapatinib-induced blockade of HER2 tyrosine kinase activity both significantly prevented EVOO polyphenols-induced cytotoxicity. EVOO polyphenols drastically depleted HER2 protein and reduced HER2 tyrosine autophosphorylation in a dose- and time-dependent manner. EVOO polyphenols-induced HER2 downregulation occurred regardless the molecular mechanism contributing to HER2 overexpression (i.e. naturally by gene amplification and ectopically driven by a viral promoter). Pre-treatment with the proteasome inhibitor MG132 prevented EVOO polyphenols-induced HER2 depletion.ConclusionThe ability of EVOO-derived polyphenols to inhibit HER2 activity by promoting the proteasomal degradation of the HER2 protein itself, together with the fact that humans have safely been ingesting secoiridoids and lignans as long as they have been consuming olives and OO, support the notion that the stereochemistry of these phytochemicals might provide an excellent and safe platform for the design of new HER2-targeting agents.

Application and potential of capillary electroseparation methods to determine antioxidant phenolic compounds from plant food material.

Antioxidants are one of the most common active ingredients of nutritionally functional foods which can play an important role in the prevention of oxidation and cellular damage inhibiting or delaying the oxidative processes. In recent years there has been an increased interest in the application of antioxidants to medical treatment as information is constantly gathered linking the development of human diseases to oxidative stress. Within antioxidants, phenolic molecules are an important category of compounds, commonly present in a wide variety of plant food materials. Their correct determination is pivotal nowadays and involves their extraction from the sample, analytical separation, identification, quantification and interpretation of the data. The aim of this review is to provide an overview about all the necessary steps of any analytical procedure to achieve the determination of phenolic compounds from plant matrices, paying particular attention to the application and potential of capillary electroseparation methods. Since it is quite complicated to establish a classification of plant food material, and to structure the current review, we will group the different matrices as follows: fruits, vegetables, herbs, spices and medicinal plants, beverages, vegetable oils, cereals, legumes and nuts and other matrices (including cocoa beans and bee products). At the end of the overview, we include two sections to explain the usefulness of the data about phenols provided by capillary electrophoresis and the newest trends.

Effect of olive ripeness on chemical properties and phenolic composition of chetoui virgin olive oil

BACKGROUND The aim of the present work was to investigate the influence of fruit ripening on oil quality in an attempt to establish an optimum harvesting time for Chétoui olives, the second main olive variety cultivated in Tunisia. RESULTS Our results showed that many analytical parameters, i.e., peroxide value, UV absorbance at 232 and 270 nm, chlorophyll pigments, carotenoids and oleic acid contents decreased during ripening, whilst oil content and linolenic acid increased. Free acidity remained practically stable with a very slight rise at the highest maturity index. The trend of oxidative stability, total phenols and o-diphenols, showed an increase at the early stages followed by a reduction at more advanced stages of maturity. The major phenolic compounds, such as hydroxytyrosol, ligstroside aglycon, elenolic acid, acetoxy-pinoresinol and oleuropein aglycon, seemed to have the same behaviour. In the case of tyrosol, a strong decrease was observed directly related with the ripening progress. CONCLUSION On the basis of the evolution of the analytical parameters studied, the best stage of Chétoui olive fruits for oil processing seems to be at ripeness index higher than 2.0 and lower than 3.0.

Gas chromatography/atmospheric pressure chemical ionization-time of flight mass spectrometry: analytical validation and applicability to metabolic profiling.

Gas Chromatography (GC)-Mass Spectrometry (MS) with Atmospheric Pressure (AP) interface was introduced more than 30 years ago but never became a mainstream technique, mainly because of technical difficulties and cost of instrumentation. A recently introduced multipurpose AP source created the opportunity to reconsider the importance of AP ionization for GC. Here, we present an analytical evaluation of GC/APCI-MS showing the benefits of soft atmospheric pressure chemical ionization for GC in combination with a Time of Flight (TOF) mass analyzer. During this study, the complete analytical procedure was optimized and evaluated with respect to characteristic analytical parameters, such as repeatability, reproducibility, linearity, and detection limits. Limits of detection (LOD) were found within the range from 11.8 to 72.5 nM depending on the type of compound. The intraday and interday repeatability tests demonstrate relative standard deviations (RSDs) of peak areas between 0.7%-2.1% and 3.8%-6.4% correspondingly. Finally, we applied the developed method to the analysis of human cerebrospinal fluid (CSF) samples to check the potential of this new analytical combination for metabolic profiling.

Gas chromatography-atmospheric pressure chemical ionization-time of flight mass spectrometry for profiling of phenolic compounds in extra virgin olive oil.

A new analytical approach based on gas chromatography coupled to atmospheric pressure chemical ionization-time of flight mass spectrometry was evaluated for its applicability for the analysis of phenolic compounds from extra-virgin olive oil. Both chromatographic and MS parameters were optimized in order to improve the sensitivity and to maximize the number of phenolic compounds detected. We performed a complete analytical validation of the method with respect to its linearity, sensitivity, precision, accuracy and possible matrix effects. The LODs ranged from 0.13 to 1.05ppm for the different tested compounds depending on their properties. The RSDs for repeatability test did not exceed 6.07% and the accuracy ranged from 95.4% to 101.5%. To demonstrate the feasibility of our method for analysis of real samples, we analyzed the extracts of three different commercial extra-virgin olive oils. We have identified unequivocally a number of phenolic compounds and obtained quantitative information for 21 of them. In general, our results show that GC-APCI-TOF MS is a flexible platform which can be considered as an interesting tool for screening, structural assignment and quantitative determination of phenolic compounds from virgin olive oil.

Reversed-phase high-performance liquid chromatography coupled to ultraviolet and electrospray time-of-flight mass spectrometry on-line detection for the separation of eight tetracyclines in honey samples

Eight antibiotics, chlortetracycline, demeclocycline, doxycycline, methacycline, minocycline, oxytetracycline, tetracycline and rolitetracycline, were separated and quantified in Spanish honey extracts of different floral origin using a commercial RP-C18 HPLC column and two different on-line detectors (diode array and electrospray time-of-flight mass spectrometry (ESI-TOF-MS) systems). Operating a linear gradient at a flow of 2 ml min(-1) the HPLC separation of the eight antibiotics was obtained within 10 min with good peak symmetry and an acceptable resolution (2.1) for the critical band pair rolitetracycline and oxytetracycline. Values of the numbers of theoretical plates (N) were comprised between 2328 and 19448 while the limits of detection in honey were within 0.02-1.03 microg kg(-1) in the case of UV detection and 0.05-0.76 microg kg(-1) for ESI-TOF-MS detection (operating in negative mode). A recovery study was carried out by preparing some quality control samples at four levels of concentration (10, 25, 50 and 100 microg kg(-1)) and percentages between 72% and 98% were attained.

Multi-component analysis (sterols, tocopherols and triterpenic dialcohols) of the unsaponifiable fraction of vegetable oils by liquid chromatography-atmospheric pressure chemical ionization-ion trap mass spectrometry.

A simple and sensitive method for the analysis of sterols, tocopherols and triterpenic dialcohols from the unsaponifiable fraction from oil samples in a single analytical run using liquid chromatography coupled to mass spectrometry was developed. With this method, the compounds could be detected directly after dissolving the unsaponifiable fraction in acetonitrile without necessity of time-consuming sample pre-treatment or derivatization. Separation of the analytes was carried out at room temperature, by using a C18 column (5 microm i.d. 3.0 mm x 250 mm) with a linear gradient of acetonitrile/water (0.01% acetic acid) at a flow rate of 1.5 mL/min. The full scan mass spectra of the investigated compounds were measured by an ion trap mass spectrometer equipped with an APCI ion source. The optimized methodology was suitable for the identification of 23 compounds belonging to different families present in olive oil and other kinds of oils, as well as for the quantification of 15 analytes (vs. their commercial standards).