Determination of coenzyme Q10 and Q9 in vegetable oils

A new sensitive and selective method has been developed for the quantification of the total coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) concentration in vegetable oil samples. The coenzyme Q fraction is isolated by solid-phase extraction (SPE) on amino phase eluting with a mixture of heptane:ethyl ether. The organic solvent is evaporated under nitrogen, and the residue is dissolved in a mixture of acetonitrile:tetrahydrofuran and finally is analyzed by reverse-phase high-performance liquid chromatography with a mass detector. The sensitivity of the method is based on the high efficient formation of the radical anions [M (-.)] of CoQ9 and CoQ10 by negative atmospheric pressure ionization. Interferences are minimized by using mass detection of the [M (-.)] ions ( m/ z = 797.5 for CoQ9 and m/ z = 862.5 for CoQ10) in selective reaction monitoring mode ( m/ z = 797.5 --> m/ z = 779.5 and m/ z = 862.5 --> m/ z = 847.5) using a triple-quadrupole mass spectrometer. The method was successfully applied to sunflower, soybean, and rapeseed oils, with a limit of quantification of 0.025 mg/kg for both compounds.     

Antioxidants and total peroxyl radical-trapping ability of olive and seed oils.

The presence of the oxidized and reduced forms of ubiquinones Q(9) and Q(10) was determined in commercial extra virgin olive and seed oils, where the amounts of alpha- and gamma-tocopherols and beta-carotene were also quantitated. Very high concentrations of ubiquinones were found in soybean and corn oils. Furthermore, the total antioxidant capability of each oil was evaluated by measuring total radical-trapping antioxidant parameters (TRAP) in tert-butyl alcohol and using egg lecithin as the oxidizable substrate. These values decreased in the order sunflower > corn > peanut > olive; the highest TRAP, which was found in sunflower oil, was related to the very high amount of alpha-tocopherol. Olive oil, because of the low content of alpha-tocopherol, exhibited a TRAP value approximately one-third that of sunflower oil. TRAP values of corn and soybean oils, in which low amounts of alpha-tocopherol but very high contents of gamma-tocopherol and reduced ubiquinones were present, were intermediate. gamma-Tocopherol exhibited a poor ability of trapping peroxyl radicals in tert-butyl alcohol. This behavior was probably due to the effects of the solvent on the rate of hydrogen abstraction from this phenol.     

复配亚麻籽油和辅酶Q10乳液的制备及表征

亚麻籽油和辅酶Q10都具有水中溶解度低、稳定性差、生物利用度低等缺点。将亚麻籽油和辅酶Q10(coenzyme Q10,CoQ10)同时负载于乳液中,可解决两者的应用瓶颈。使用阿拉伯胶为乳化剂,采用高压均质法制备复配亚麻籽油和CoQ10乳液。采用动态光散射、透射电子显微镜、体外模拟消化、体外释放、稀释稳定性、冻融稳定性、离子强度稳定性、光稳定性和加速氧化稳定性方法对所制备乳液的理化性质进行表征。结果显示,制备的乳液平均粒径为(284±5.6) nm,多分散指数(polydispersity index,PDI)为0.112±0.025,为均匀分散的球形液滴。制备的乳液在模拟小肠液中消化,和亚麻籽油、CoQ10混悬液相比,乳化后亚麻籽油的消化速率和CoQ10的生物可给率明显提高。乳液中CoQ10的释放表现出缓释效果。制备的乳液具有较好的稀释和冻融稳定性。Na^+和Ca^2+会造成乳液Zeta电位的下降,对乳液稳定性影响较大。乳液载体化后CoQ10的光稳定性得到了提高。CoQ10对亚麻籽油具有较好的保护作用。     

Antioxidant effect of two virgin olive oils depends on the concentration and composition of minor polar compounds

In vitro studies show that some individual minor polar phenolic compounds (MPC) present in virgin olive oil prevent oxidation of human low-density lipoproteins (LDL), but few data are available on the antioxidant effect of whole oil extract. Thus, whole virgin olive extracts were studied to determine whether they maintain the antioxidant activity and whether this last is linked to MPC composition of a single virgin oil. Using HPLC-DAD the MPC content in Taggiasca and Seggianese virgin olive oils was measured. Taggiasca oil was less rich in total MPC (208.5 mg/L) than Seggianese oil (441.9 mg/L). In addition, the major compounds of Taggiasca oil were lignan derivatives, whereas the major compounds in Seggianese oils were secoiridoid derivatives. Moreover, Taggiasca oil was practically free of 5-hydroxytyrosol and 5-hydroxytyrosol derivatives, deacetoxy-oleuropein aglycone and oleuropein aglycone. The antioxidant activity of the oils on human LDL was evaluated by measuring malondialdehyde and conjugate diene generation induced by copper ions. In both tests, the oil extracts dose-dependently reduced malondialdehyde and conjugate diene generation. Moreover, antioxidant potency correlated with total MPC; thus, Seggianese extract was more active. The two oils differed quantitatively and qualitatively, and these differences influenced their biological activities; thus clinical trials focused on studying the effects of olive oils should specify the oils used.     

Comparison of volatile aldehydes present in the cooking fumes of extra virgin olive, olive, and canola oils

Emissions of low molecular weight aldehydes (LMWAs) from deep-frying of extra virgin olive oil, olive oil, and canola oil (control) were investigated at two temperatures, 180 and 240 degrees C, for 15 and 7 h, respectively. The oil fumes were collected in Tedlar bags and then analyzed by gas chromatography-mass spectrometry. Seven alkanals (C2-C7 and C9), eight 2-alkenals (C3-C10), and 2,4-heptadienal were found in the fumes of all three cooking oils. The generation rates of these aldehydes were found to be dependent on heating temperature, showing significant increases with increases in temperature. The LMWA emissions from both kinds of olive oils were very similar and were lower than those observed from canola oil under similar conditions. These results suggest that frying in any type of olive oil, independent of its commercial category, will effectively decrease the generation of volatile aldehydes in the exhaust. This fact is important because less expensive refined olive oil is usually used for deep-frying operations, whereas extra virgin olive oil is usually used as salad dressing.     

Coenzyme Q9 provides cardioprotection after converting into coenzyme Q10

Coenzyme Q10 (CoQ10) has been extensively studied as adjunctive therapy for ischemic heart disease, and its cardioprotective ability is well-established. The mitochondrial respiratory chain contains several coenzymes, including CoQ1, CoQ2, CoQ4, CoQ6, CoQ7, CoQ8, CoQ9, and CoQ10. It is not known whether other CoQs, especially CoQ9, is equally cardioprotective as CoQ10. The present study was designed to determine if CoQ 9 could protect guinea pig hearts from ischemia reperfusion injury. Guinea pigs were randomly divided into three groups: groups I and II were fed CoQ 9 and CoQ10, respectively, for 30 days while group III served as control. After 30 days, the guinea pigs were sacrificed and isolated hearts were perfused via working mode were subjected to 30 min ischemia followed by 2 h of reperfusion. Cardioprotection was assessed by evaluating left ventricular function, ventricular arrhythmias, myocardial infarct size, and cardiomyocyte apoptosis. Samples of hearts were examined for the presence of CoQ9 and CoQ10. The results demonstrated that both CoQ9 and CoQ10 were equally cardioprotective, as evidenced by their abilities to improve left ventricular performance and to reduce myocardial infarct size and cardiomyocyte apoptosis. High performance liquid chromatographic (HPLC) analysis revealed that a substantial portion of CoQ9 had been converted into CoQ10. The results indicate that CoQ9 by itself, or after being converted into CoQ10, reduced myocardial ischemia/reperfusion-induced injury.     

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (β-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.     

Rapid quantitative assessment of the adulteration of virgin olive oils with hazelnut oils using Raman spectroscopy and chemometrics

The authentication of extra virgin olive oil and its adulteration with lower-priced oils are serious problems in the olive oil industry. In addition to the obvious effect on producer profits, adulteration can also cause severe health and safety problems. A number of techniques, including chromatographic and spectroscopic methods, have recently been employed to assess the purity of olive oils. In this study Raman spectroscopy together with multivariate and evolutionary computational-based methods have been employed to assess the ability of Raman spectroscopy to discriminate between chemically very closely related oils. Additionally, the levels of hazelnut oils used to adulterate extra virgin olive oil were successfully quantified using partial least squares and genetic programming.     

Effect of different olive oils on bile excretion in rats fed cholesterol-containing and cholesterol-free diets

The mechanism of the hypocholesterolemic effect of olive oils was investigated in 60 Wistar rats adapted to cholesterol-containing and cholesterol-free diets. The rats were divided in six diet groups of 10. The control group was fed only basal diet (BD), which contained wheat starch, casein, cellulose, and mineral and vitamin mixtures. For the five other groups, 10 g/100 g virgin (virgin group) or lampante (lampante group) olive oils, 1 g/100 g cholesterol (chol group), or both cholesterol and oil (chol/virgin and chol/lampante groups) were added to the BD. The experiment lasted 4 weeks. Before and after the experiment the bile was collected, and its flow and biliary bile acids and cholesterol concentrations were registered. Plasma lipids, liver cholesterol, plasma antioxidative potential (TRAP), fecal output, fecal bile acids, and fecal cholesterol excretion were measured. Groups did not differ before the experiment. After the experiment significant hypocholesterolemic and antioxidant effects were registered mainly in groups of rats fed cholesterol-containing diets supplemented with both olive oils (chol/virgin and chol/lampante). Significant increases in the bile flow and in the bile cholesterol and bile acids concentrations were observed (19.2% and 16.9%, 30.5% and 18.2%, and 79.6% and 45.6% for the chol/virgin and chol/lampante groups, respectively). Also, significant increases of the fecal output and fecal excretion of bile acids and cholesterol in rats of these groups were found. In conclusion, olive oils positively affect plasma lipid metabolism. The hypocholesterolemic effect of olive oils is genuine and is most likely mediated through increases in bile flow and biliary cholesterol and bile acids concentrations and subsequent increases in their fecal excretion.     

Determination of the diglyceride content in greek virgin olive oils and some commercial olive oils by employing (31)P NMR spectroscopy

In this study, the diglyceride contents of 96 samples of virgin olive oils from the regions of Crete, Lesvos, Messinia, Pilion, Zakynthos, Halkidiki, and Ilia, 15 samples of commercial extra virgin and pure olive oils, and 3 samples each of refined olive oils and pomace oils were determined by a facile method introduced in a previous publication. This method is based on the phosphitylation of the free hydroxyls of the diglycerides with 2-chloro-4,4,5,5-tetramethyldioxaphospholane and the integration of the appropriate peaks in the (31)P NMR spectra. This preliminary study showed interesting trends in the diglyceride content of the virgin olive oils from the various regions of Greece that can be used as simple criteria to assess the olive oil characteristics. Analysis of variance has been carried out for the diglyceride content of each region in an attempt to detect possible differences in the diglyceride levels among the various regions. Finally, the relationship between the ratio of 1,2-diglycerides to the total amount of diglycerides and the total amount of diglycerides has been used to monitor the quality of virgin olive oils, commercial olive oils, refined olive oils, and pomace oils.     

Antithrombotic lipid minor constituents from vegetable oils. Comparison between olive oils and others

Many epidemiological studies suggest that vegetable oils and especially olive oil present a protective effect against atherosclerosis. In this study, total lipids (TL) of Greek olive oils and seed oils of four kinds, namely, soybean, corn, sunflower, and sesame oil, were separated into total polar lipids (TPL) and total neutral lipids (TNL) via a novel extraction procedure. TPL and TNL of olive oil were fractionated by HPLC for further study. Each lipid fraction from HPLC separation along with TL, TPL, and TNL lipid samples from oils were tested in vitro for their capacity to induce or to inhibit washed rabbit platelet aggregation. Comparison between olive and seed oils supports the superiority of olive oil as high levels of platelet activating factor (PAF) antagonists have been detected, mainly in TPL. In addition, the structure of the most active fraction from olive oil was elucidated, as a glycerol-glycolipid. Because it has already been reported that PAF plays a pivotal role in atherogenesis, the existence of PAF agonists and antagonists in vegetable oils may explain their protective role against atherosclerosis.     

Optimization in the preparation of coenzyme Q10 nanoliposomes

The optimal formulation of coenzyme Q10 (CoQ10) nanoliposomes and the feasibility of production in a pilot scale were investigated. The nanoliposomes were prepared by ethanol injection and sonication techniques for a desired vesicle size in the laboratory. Optimization of formulation in the preparation of CoQ10 nanoliposomes was achieved by an orthogonal array design. The best formulation was found to be phospholipid/CoQ10/cholesterol/Tween 80 (2.5:1.2:0.4:1.8, w/w) with phosphate buffer solution (pH 7.4, 0.01 M) as the hydration media. The z-average diameter (D(z)) was about 68 nm. The encapsulation efficiency was greater than 95% with a retention ratio higher than 90% and a particle size change lower than 10% after storage at 4 degrees C in the dark for 90 days. CoQ10 incorporation resulted in a dramatic increase of the microviscosity of nanoliposomes and inhibited the peroxidation of phospholipid. The D(z) of CoQ10 nanoliposomes produced in a pilot scale was about 67 nm. Results suggest that the technology developed by this investigation is practical to produce the CoQ10 nanoliposomes with the expected encapsulation quality and stability not only in the laboratory but also in a pilot scale.     

Interpretation of Fourier transform Raman spectra of the unsaponifiable matter in a selection of edible oils.

The unsaponifiable matter of edible oils is a source of information for their characterization and authentication. FT-Raman spectroscopy has been applied with success to the determination of the spectra of the unsaponifiable matter of varietal olive oils as well as other refined and crude edible oils. The spectra of the major unsaponifiable series of compounds (squalene, sterolic, and terpenic alcoholic fractions), together with beta-carotene and lutein, have been used to explain the most prominent bands found in the spectra of the unsaponifiable matter of 15 edible oil samples. The order of the scattering intensities of the varietal olive oils agrees with the results obtained by chromatography. An unsupervised multivariate statistical analysis of selected bands points out differences between olive oils and the other seed oils and also among varietal virgin olive oils.     

Comparison of the antioxidant activities of extra virgin olive oils

The phenol content and antioxidant activity of extra virgin olive oils (EVOOs) differing in their origins and degradation degrees were studied. The o-diphenolic compounds typical of olive oil, namely, the oleuropein derivatives hydroxytyrosol (3',4'-dihydroxyphenylethanol, 3',4'-DHPEA), the dialdehydic form of elenolic acid linked to 3',4'-DHPEA (3',4'-DHPEA-EDA), and an isomer of oleuropein aglycon (3',4'-DHPEA-EA), were analyzed by HPLC. The antioxidant activity was studied by (a) the xanthine oxidase (XOD)/xanthine system, which generates superoxide radical and hydrogen peroxide; (b) the diaphorase (DIA)/NADH/juglone system, which generates superoxide radical and semiquinonic radical; and (c) the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) test. Results showed that EVOOs with a low degradation level (as evaluated by acidity, peroxide number, and spectroscopic indices K(232), K(270), and deltaK according to the EU Regulation) had a higher content of 3',4'-DHPEA-EDA and a lower content of 3',4'-DHPEA than oils having intermediate and advanced degradation levels. EVOOs with a low degradation degree were 3-5 times more efficient as DPPH scavengers and 2 times more efficient as inhibitors of the XOD-catalyzed reaction than oils with intermediate and advanced degradation levels. The DIA-catalyzed reaction was inhibited by EVOOs having low or intermediate degradation levels but not by the most degraded oils.     

Alkyl esters of fatty acids a useful tool to detect soft deodorized olive oils

Fatty acid alkyl esters (FAAEs) are a family of natural neutral lipids present in olive oils and formed by esterification of free fatty acids (FFAs) with low molecular alcohols. Inappropriate practices during the olive oil extraction process and bad quality of the olive fruits promote their formation. Quantification can be done by isolation with a silica gel solid phase extraction cartridge followed by analysis on a gas chromatograph equipped with a programmed temperature vaporizer injector using a polar capillary column. The application of the method to more than 100 Spanish olive oils from different categories, varieties, and geographical origin allowed for establishing the average content of FAAEs and distinguishing the Spanish protected denomination of origin (PDO) and extra virgin olive oils from other categories of olive oils. Those other categories of oils can be subjected to a mild refining process, which leads to blending with extra virgin olive oils. Studies on low quality oils subjected to mild refining showed that FAAEs remain after that process. Thereby, blends of extra virgin olive and mildly refined low quality olive oils can be detected by their alkyl ester concentrations.     

Comparison of static headspace, headspace solid phase microextraction, headspace sorptive extraction, and direct thermal desorption techniques on chemical composition of French olive oils

Static headspace (SHS), headspace solid phase microextraction (HS-SPME), headspace sorptive extraction (HSSE), and direct thermal desorption (DTD) were applied to the analysis of four French virgin olive oils from Corsica. More than 60 compounds were isolated and characterized by GC-RI and GC-MS. SHS was not suited to the characterization of olive oil volatile compounds because of low sensitivity. The SPME and HSSE techniques were successfully applied to olive oil headspace analysis. Both methods allow the characterization of volatile compounds (mainly C(6) aldehydes and alcohols), which contribute significantly to the "green" flavor note of virgin olive oils. The PDMS stir bar showed a higher concentration capacity than a DVB/CAR/PDMS SPME fiber due to the higher volume of polymeric coating. DTD was a very good tool for extracting volatile and especially semivolatile compounds, such as sesquiterpenes, but requires a significant investment like that for HSSE. Finally, SPME may be a more appropriate technique for routine quality control due to its operational simplicity, repeatability, and low cost.     

Qualitative and semiquantitative analysis of phenolic compounds in extra virgin olive oils as a function of the ripening degree of olive fruits by different analytical techniques

Capillary electrophoresis (CE) can be effectively used as a fast screening tool to obtain qualitative and semiquantitative information about simple and complex phenolic compounds of extra virgin olive oil. Three simple phenols (tyrosol, hydroxytyrosol, and vanillic acid), a secoiridoid derivative (deacetoxy oleuropein aglycon), and two lignans (pinoresinol and acetoxypinoresinol) were detected as the main compounds in extra virgin olive oils by high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE). Spectrophotometric indices, radical scavenging activity, and oxidative stability of extra virgin olive oil samples obtained from olives hand-picked at different ripening degrees were statistically correlated with the CZE and HPLC quantification. The concentration of phenols in extra virgin olive oil decreased with ripeness of olive fruits. The high correlations found between CZE and the other analytical results indicate that CE can be applied as a rapid and reliable tool to routinely determine phenolic compounds in extra virgin olive oils.     

Comparison of the radical scavenging potential of polar and lipidic fractions of olive oil and other vegetable oils under normal conditions and after thermal treatment

The antioxidant activity (IC(50)) of extra virgin olive oil (EVOO), commercial olive oil, and other vegetable oils (soybean, sunflower, and corn oil) was determined by UV-vis and by electron paramagnetic resonance (EPR) spectroscopy of the stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). Also, we studied the antioxidant activity of the methanol soluble phase (methanolic, MF) and the nonsoluble phase (lipidic, LF) of oils by the same methods. Similarly, we studied the effect of heating on the antioxidant activity at 160 and 190 degrees C. Also, the MF, containing the polyphenolic substances, was used for measurements of the radical scavenging capacity toward the most important oxygen free radicals, superoxide anion (O(2)(*)(-)) and hydroxyl (HO(*)) radicals. Results showed that soybean oil and EVOO had the highest antioxidant potential and thermal stability. In the case of soybean oil, the antioxidant capacity is the result of its high content of gamma- and delta-tocopherols (with the highest antioxidant capacity and thermostabilities), whereas in EVOO, the antioxidant potential is the result of the combination of specific antioxidant polyphenols, which are acting additionally as effective stabilizers of alpha-tocopherol. The high content of EVOO in tyrosol, hydrotyrosol, and oleuropein and other polyphenolics with radical scavenging abilities toward superoxide anion and hydroxyl radical suggests that olive oil possesses biological properties that could partially account for the observed beneficial health effects of the Mediterranean diet.     

Influence of deep frying on the unsaponifiable fraction of vegetable edible oils enriched with natural antioxidants

The influence of deep frying, mimicked by 20 heating cycles at 180 °C (each cycle from ambient temperature to 180 °C maintained for 5 min), on the unsaponifiable fraction of vegetable edible oils represented by three characteristic families of compounds (namely, phytosterols, aliphatic alcohols, and triterpenic compounds) has been studied. The target oils were extra virgin olive oil (with intrinsic content of phenolic antioxidants), refined sunflower oil enriched with antioxidant phenolic compounds isolated from olive pomace, refined sunflower oil enriched with an autoxidation inhibitor (dimethylpolysiloxane), and refined sunflower oil without enrichment. Monitoring of the target analytes as a function of both heating cycle and the presence of natural antioxidants was also evaluated by comparison of the profiles after each heating cycle. Identification and quantitation of the target compounds were performed by gas cromatography-mass spectrometry in single ion monitoring mode. Analysis of the heated oils revealed that the addition of natural antioxidants could be an excellent strategy to decrease degradation of lipidic components of the unsaponifiable fraction with the consequent improvement of stability.     

Protective effect of the phenolic fraction from virgin olive oils against oxidative stress in human cells

This paper reports the protective effect of the phenolic fraction extracted from extra virgin olive oils (OOPEs) against the cytotoxic effects of reactive oxygen species in human erythrocytes and Caco-2 cells, employed as model systems. Pretreatment of cells with various OOPEs, indeed, provides a remarkable protection against oxidative damages: this effect was strictly dependent on the o-diphenolic content of the extracts. Moreover, the protective effects observable in cellular systems were compared with in vitro antioxidant properties, measured by using the FRAP (ferric reducing/antioxidant power) assay; the reducing ability of OOPEs strictly parallels their o-phenolic content. The linear relationship demonstrated between biological effects and antioxidant capacity measured by the FRAP assay allows us to propose the use of this rapid colorimetric method in assessing and certifying the antioxidant power of extra virgin olive oil.