Co-Microencapsulation of Cushuro (Nostoc sphaericum) Polysaccharide with Sacha Inchi Oil (Plukenetia huayllabambana) and Natural Antioxidant Extracts.

Cushuro (Nostoc sphaericum) polysaccharide was used to co-microencapsulate sacha inchi oil, natural antioxidant extracts from the oleoresin of charapita chili peppers (Capsicum frutescens L.) and grape orujo (Vitis vinifera L.). Encapsulation efficiency, moisture, particle size, morphology, oxidative stability, shelf-life, solubility, essential fatty acid profile, sterol content and antioxidant capacity were evaluated. The formulations with grape orujo extract showed higher oxidative stability (4908 ± 184 h), antioxidant capacity (4835.33 ± 40.02 µg Trolox/g ms), higher phenolic contents (960.11 ± 53.59 µg AGE/g ms) and a smaller particle size (7.55 µm) than the other formulations, as well as good solubility and a low moisture content. Therefore, grape orujo extracts can be used as natural antioxidants. The fatty acid composition (ω-3) remained quite stable in all the formulations carried out, which also occurred for sterols and tocopherols. In combination with gum arabic, grape orujo extract offered oxidative protection to sacha inchi oil during the first week of storage.

Co-Microencapsulation of Sacha Inchi (Plukenetia huayllabambana) Oil with Natural Antioxidants Extracts.

Sacha inchi (Plukenetia huayllabambana) oil was co-microencapsulated with natural antioxidant extracts (NAE), such as camu-camu (Myrciaria dubia (HBK) Mc Vaugh) fruit, Añil variety Andean potato (Solanum tuberosum andigenum, and elderberry fruit (Sambucus peruviana). Gum Arabic and the ternary combination of gum Arabic (GA) + maltodextrin (MD) + whey protein isolate (WPI) at different formulations were used as coating materials for the encapsulation process using spray-drying. The moisture content, particle size distribution and morphology, total phenolic content, antioxidant activity, fatty acid and sterol composition, oxidative stability, and shelf-life were evaluated. Co-microcapsules of sacha inchi (P. huayllabambana) oil with camu camu skin extract (CCSE) at 200 ppm encapsulated with GA + MD + WPI had the highest total polyphenol content (4239.80 µg GAE/g powder), antioxidant activity (12,454.00 µg trolox/g powder), omega-3 content (56.03%), ß-sitosterol (62.5%), greater oxidative stability (Oxidation Onset temperature of 189 °C), higher shelf-life (3116 h), and smaller particle sizes (6.42 µm). This research enhances the knowledge to obtain microcapsules containing sacha inchi (P. huayllabambana) oil with natural antioxidant extracts that could be used for the development of functional foods. Further research is needed to study the potential interactions and their influence between the bioactive components of the microcapsules and the challenges that may occur during scale-up to industrial production.

The Mineral Oil Hydrocarbon Paradox in Olive Pomace Oils.

The aim of this work was to understand the actual content of mineral oil hydrocarbons (MOH) in olive pomace oil in order to contribute to the monitoring requested by EFSA for the food groups making a relevant impact on human background exposure. Such information will complement both the information inferred from the limits established by the EU and the interpretation of the coming toxicological risk assessment. At the same time, the origin of such a group of compounds is discussed. From the raw material to the commercial product, olive pomace oils were sampled and analyzed at different points and/or conditions. Through the ultimate online HPLC-GC-FID system, we gathered information on the MOH concentrations and molecular mass profiles (C-fractions), and through GCxGC-TOF/MS, we identified the key structures that prove the innocuousness of the mineral oil aromatic hydrocarbon (MOAH) fraction. Our approaches provided chromatographic signals on the C10-C50 range, rendering 33-205 mg/kg mineral oil saturated hydrocarbon (MOSH) and 2-55 mg/kg MOAH in the commercial product. The results confirmed that the C25-C35 cut is the main fraction to which humans are exposed via olive pomace oil, showing concentrations highly dependent on the extraction process. Moreover, the identification of the main MOAH groups showed that in olive pomace oil, mainly 1- and 2-ring species were present, being virtually free of the carcinogenic 3-7 ring aromatics.

Fatty acid ethyl esters (FAEE) in virgin olive oil: A shorter and full validated approach as an alternative to the EU Official Method.

In this work a SPE/GC-FID method, incorporating the use of a 1-g silica cartridge, for the determination of FAEE in olive oils is presented. The procedure has been fully validated, initially 'in-house' and subsequently by an international validation study involving sixteen laboratories from Europe, the United States of America, and China. Key performance parameters of the method are: (1) Linearity in the 10-134 mg/kg range (R2 > 0.999), (2) LOD and LOQ < 0.5 mg/kg, (3) RSDr < 10%, (4) RSDR < 20% (for 4 out of 5 test materials). In addition, the method has been demonstrated to provide equivalent results to the Official Method (Commission Regulation 2568/91) while providing advantages in terms of reductions in time and solvents and ease of automation. In fact, the proposed protocol requires 30 mL solvents and takes 1.5 h per determination instead of the 350 mL and 6 h needed in the UE Official Method.

Fatty Acid Ethyl Esters in Virgin Olive Oils: In-House Validation of a Revised Method.

The content of fatty acid ethyl esters (FAEEs) is one of the quality parameters to define if an olive oil can be classified as extra virgin as these compounds are considered markers for virgin olive oils obtained from poor-quality olives. In addition, FAEEs can also be indirect markers to detect soft deodorization treatment. In this study, an off-line HPLC-GC-FID method for determination of FAEEs is presented, revising the preparative step and the GC injector required by the official method (EU Reg. 61/2011). After optimization, the method was validated in-house by analyzing several parameters (linearity, limit of detection LOD, limit of quantification LOQ, robustness, recovery, precision, and accuracy) to determine its effectiveness. Linearity was measured in the 2.5-50 mg/L range; furthermore, intra-day and inter-day precision values were lower than 15%, while the LOD and LOQ were lower than 1 and 1.5 mg/kg, respectively, for all compounds considered. The main advantages of this revised protocol are: (i) significant reduction in time and solvents needed for each analytical determination; (ii) application of HPLC as an alternative to traditional LC, carried with manually packed glass columns, thus simplifying the separation step.

Pyropheophytin a in Soft Deodorized Olive Oils.

Mild refined olive oil obtained by neutralization and/or by soft deodorization at a low temperature and its blending with extra virgin olive oil (EVOO) is not allowed and is difficult to detect. Chlorophyll derivatives, pheophytins and pyropheophytin, and their relative proportions were proposed as parameters to detect such processes. The objective of this study is to determine changes in EVOO, in terms of pheophytins and pyropheophytin, occurring after several well-controlled mild refining processes. The changes on those chlorophyll pigments due to the processes depend on the temperature, stripping gas, acidity and oil nature. The data obtained show that, at temperatures below 100 °C, the rate at which pyropheophytin a is formed (Ra) is lower than the rate at which pheophytins a+a' disappear (Ra+a'). As a consequence, the Ra+a' and Ra ratios are considered to be directly linked to pheophytins a+a' decrease instead of to pyropheophytin a formation. Stripping gas very slightly affects the transformation of the chlorophyll pigments; actually both acidity and N2 enhance the increment in the Ra+a' and Ra ratios. In relation to the oil nature, the higher the initial pheophytin a+a' content, the higher the increase in the Ra+a' and Ra relations.

Olive oil mixtures. Part one: Decisional trees or how to verify the olive oil percentage in declared blends.

The commercialization of declared blends of olive oil and seed oil is something long approved by the European Union. There, the olive oil percentage must be at least 50% if the producer aims to advertise its presence on the front label, i.e., somewhere other than in the ingredients list. However, the Regulation did not propose any method to verify such proportion. For this purpose, we recommend the use of decisional trees, being the parameters under study those in which the greatest differences between olive and seed oils are shown: triacylglycerols, acyclic saturated hydrocarbons, free sterols, and tocopherols. In this way, to guarantee the presence of olive oil at 50%: i) palmitodiolein must be above 11-15%; ii) the ß/γ-tocopherol ratio must be below 2.4; iii) the alkane sum C21-C25 should be higher than 3.5-6%; and iv) the total sterol content cannot surpass 2400 mg/kg.

Saturated hydrocarbon content in olive fruits and crude olive pomace oils.

Olive fruits contain an n-alkane series of saturated hydrocarbons mainly in the pulp. Lower amounts of a complex mixture of paraffins, unresolved by gas chromatography (UCM--unresolved complex mixture), have been found in cuticle, stone (woody shell and seed), olive leaves, and talc used as an aid to olive oil extraction. The amounts of both kinds of hydrocarbons are related to the olive cultivar and are transferred to oils in a proportion depending on the oil-obtaining process (centrifugation or solvent extraction). In olive oil obtained by centrifugation, only n-alkanes were detected. However, in olive oil extracted by second centrifugation, small amounts of UCM paraffins were detected together with the n-alkanes. Olive pomace oils showed a very variable content of both types of hydrocarbons according to the different obtaining process, such as double centrifugation, solvent extraction or centrifugation followed by solvent extraction. 'White mineral oil' used in oil extraction machinery is the source of the high concentrations of UCM paraffins found in some olive and olive pomace oils. In the case of second centrifugation olive oil, a maximum limit of 50 mg kg(-1) of UCM is suggested, whereas in the case of crude olive pomace oil, it amounts to 250 mg kg(-1) plus an additional minimum of 1.0 for the n-alkanes/UCM ratio.

Comparative study of phytosterol derivatives in monovarietal olive oils.

Plant sterols and their derivatives are minor compounds that have been extensively studied in vegetable oils, mainly in olive oil, where they are closely related with its identity. The objective of this work is to determine the content of free and esterified steryl glucosides and their profiles in olive oil in relation to different geographical situation of olive orchards, cultivar, farming modality, and sampling time. The orchards under study were located in the outer ring of the submetropolitan area of Madrid (Spain), where olives from Cornicabra, Manzanilla Cacereña, Manzanilla Castellana, and Picual varieties were grown under traditional and organic modes, and harvested in four different samplings. Conclusions state that cultivar, farming mode, and light exposure do not have outstanding effects, whereas pedoclimate might affect the steryl glucoside presence in a substantial way. Further studies are being carried out presently in order to confirm such statement. Also glucoside derivative profiles are discussed, and reasons for differences with results in previous studies pointed out.

Analysis of methanol and ethanol in virgin olive oil.

This work provides a short and easy protocol that allows the analysis of both methanol and ethanol in the static headspace of olive oil. The procedure avoids any kind of sample pre-treatment beyond that of heating the oil to allow a maximum volatile concentration in the headspace of the vials. The method's LOD is 0.55 mg kg(-1) and its LOQ is 0.59 mg kg(-1). Advantages of this method are:•Simultaneous determination of methanol and ethanol (the pre-existing Spanish specification UNE-EN 14110 only analyses methanol).•No need of equipment modifications (standard split injectors work perfectly). Use of a highly polar capillary GC column, leading in most cases to chromatograms in which only three dominant peaks are present - methanol, ethanol, and propanol (that is extremely positive for easy interpretation of results).•Use of an internal standard (1-propanol) to determine the concentration of the analytes, reducing the presence of error sources.

On the glucoside analysis: simultaneous determination of free and esterified steryl glucosides in olive oil. Detailed analysis of standards as compulsory first step.

This work covers two important gaps in the field of micronutrient databases: herein we describe a short and easy protocol that allows the analysis of both free and esterified steryl gulcosides in olive oil. By utilising accurate quantitative methods we achieve a better understanding of olive oil composition and health promoting properties. The procedure consists of isolating the fraction of interest through solid phase extraction, and using gas chromatography-flame ionisation detection for both identification and quantification of the derivatised species. Additionally, mass-spectrometry detection has been utilised for confirming the identity of the individual esterified steryl glucosides in some cases. The method's limit of detection has been set at 0.37mg/kg for each free steryl glucoside and 0.20mg/kg for each esterified steryl glucoside, whereas the recoveries are around 96% and 77%, respectively. Finally, we provide a complete analysis of the commercial standard for esterified steryl glucosides, since such information was not yet available.

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.

Sources of contamination by polycyclic aromatic hydrocarbons in Spanish virgin olive oils.

The presence of polycyclic aromatic hydrocarbons (PAHs) in virgin olive oils results from contamination on olive skins and the oil itself during processing. Determination of nine PAHs was carried out by isolation of the hydrocarbon fraction and subsequent clean-up by solid phase extraction, followed by RP-HPLC analysis using a programmable fluorescence detector. Contamination of olive skins depends directly on environmental pollution levels and inversely on fruit size. In the oil mill, PAHs levels were increased by contamination from combustion fumes during the extraction process. Other procedures, such as washing or talc addition during extraction, did not affect PAHs levels. High concentrations of PAHs were only found as a consequence of accidental exposure to contamination, such as direct contact of olives with diesel exhaust and oil extraction in a polluted environment.