Toward a Harmonized and Standardized Protocol for the Determination of Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil (VOO). The Pros of a Fit for the Purpose Ultra High Performance Liquid Chromatography (UHPLC) Procedure.

Τoward a harmonized and standardized procedure for the determination of total hydroxytyrosol and tyrosol content in virgin olive oil (VOO), the pros of a recently published in house validated ultra high performance liquid chromatography (UHPLC) protocol are discussed comparatively with those of other procedures that determine directly or indirectly the compounds hosted under the health claim on "olive oil polyphenols" (EC regulation 432/2012). Authentic VOOs were analyzed with five different liquid chromatographic separation protocols and 1H-NMR one in five different laboratories with expertise in VOO phenol analysis within three months. Data comparison indicated differences in absolute values. Method comparison using appropriate tools (Passing-Bablok regression and Bland Altman analyses) for all protocols vs. the UHPLC one indicated slight or statistically significant differences. The results were also discussed in terms of cost effectiveness, detection means, standard requirements and ways to calculate the total hydroxytyrosol and tyrosol content. Findings point out that the in-house validated fit for the purpose UHPLC protocol presents certain pros that should be exploited by the interested parties. These are the simplicity of sample preparation, fast elution time that increase the number of samples analyzed per day and integration of well-resolved peaks with the aid of only two commercially available external standards. Importance of correction factors in the calculations is stressed.

Italian and Spanish commercial tomato sauces for pasta dressing: study of sensory and head-space profiles by Flash Profiling and solid-phase microextraction-gas chomatography-mass spectrometry.

BACKGROUND: The sensory and head-space profiles of Italian and Spanish commercial tomato sauces were investigated. The Flash Profiling method was used to evaluate sensory characteristics. Samples within each set were ranked according to selected descriptors. One hundred volatile compounds were identified by solid-phase microextraction-gas chomatography-mass spectrometry. RESULTS: For Italian samples, the sensory notes of basil/aromatic herbs, acid and cooked tomato were among those perceived most by the assessors, whereas, in Spanish samples, the sensory attributes of garlic/onion and onion/sweet pepper and, in Italian samples, cooked tomato were among those found most frequently. Data were elaborated using multivariate statistical approaches and interesting correlations were observed among the different sensory attributes and related volatile compounds. CONCLUSION: Spanish samples were characterized by the highest content of volatiles linked to the thermal treatment of tomatoes and to raw and sautéed garlic and onion, whereas the Italian samples were characterized by terpenic compounds typical of basil and volatile molecules derived from fresh tomato. These results confirm the influence of both formulation and production processes on the aromatic profile (sensory attributes and volatile compounds) of tomato products, which is probably related to the different eating habits and culinary traditions in Italy and Spain. © 2016 Society of Chemical Industry.

Olive oil enriched in lycopene from tomato by-product through a co-milling process.

The aim of this investigation was to produce an olive oil (OO) naturally enriched with antioxidants, recovering carotenoids, in particular lycopene, using an industrial by-product of tomato seeds and skin. For this purpose, a technological process in a low-scale industrial plant to co-mill olives and tomato by-product in de-frosted or freeze-dried forms was applied and studied with respect to control samples. Preliminary results obtained from two different experiments were carried out by 40 kg of cultivar Correggiolo olives and 60 kg of olive blends from different cultivars. In both the experiments, the co-milling showed significant enrichment in carotenoids, especially in lycopene (mean values of 5.4 and 7.2 mg/kg oil from defrosted and freeze-dried by-products, respectively). The experimental results demonstrated the possibility to obtain a new functional food naturally enriched in antioxidant compounds, which might be marketed as "OO dressing enriched in lycopene" or "condiment produced using olives and tomato by-product".

Characterization of olive mill wastewater fractions treatment by integrated membrane process.

BACKGROUND: Up to now, the management of olive mill wastewaters, a three-phase mill by-product, remains an unsolved problem, in particular for those regions where huge quantities of vegetable water are produced. Olive mill wastewaters were therefore treated to evaluate the characteristics of permeate and retentate fractions produced by an integrated membrane system working at two different volume concentration factors. RESULTS: The effect of two membrane-based filtration steps (microfiltration and nanofiltration) on the content of chemical oxygen demand, dry matter, sensory quality, phenolic compounds and antioxidant activity of permeate and retentate samples was evaluated. Furthermore, the effect of two different volume concentration factors (VCF), in the nanofiltration step, were investigated. At high VCF values, the total phenolic content in the retentate fraction was found to be 3.7-fold higher than the starting one, while the reduction of chemical oxygen demand in the permeate fraction was greater than 97% also at lower VCF values. CONCLUSION: Each filtration step has provided useful information concerning the utility and appropriateness of the processes chosen, suggesting a sustainable hypothesis of 'normal industrial practice' that can be included in current processes of oil extraction, in order to purify water and recover phenolic compounds with high added value.

Organic and conventional nonflavored yogurts from the Italian market: study on sensory profiles and consumer acceptability.

BACKGROUND: The sensory properties of food products are an important success factor, especially in the organic market, where many producers and distributors of organic food claim superior taste for their products compared to the conventional alternative. For this reason consumer expectations and preferences, as well as the sensory properties of conventional and organic yogurt, have to be investigated in depth. In this work, the sensory profiling and consumer data of six nonflavored organic and conventional Italian yogurts were elaborated. Some results on the data segmentation (heavy and light users of organic food) and on the effect of information on liking (blind and labeled test) were obtained. Multivariate analysis was carried out to study how the sensory characteristics of 'natural yogurts' drive consumer liking. RESULTS: Consumers' preferences were oriented towards a creamy mouthfeel and smooth visual appearance and for a less acid and fresh taste. In particular, a conventional yogurt was the least accepted, because it was not creamy enough. This paper shows there is room to improve unflavored yogurt to better meet consumer expectations. CONCLUSION: Sensory profiling did not allow a distinction in odor/taste/texture between organic and conventional samples. However, three of four organic samples were in the region of highest consumer acceptability, fitting well consumers' preference. There was no clear tendency that heavy or light users scored differently in the blind and labeled tests regarding overall liking but, for all, the most liked conventional yogurt scored higher when labeled as organic.

Evaluation of Hemp Seed Oils Stability under Accelerated Storage Test.

The interest in hemp seed oil has recently increased, due to the latest regulations which allow its use as food. Hemp seed oil is characterized by a high content of polyunsaturated fatty acids, which are highly prone to oxidation. Accelerated thermal oxidation (60 °C, 18 days) has been applied to nine types of cold-pressed hemp seed oils to monitor the evolution of the samples during oxidative deterioration. The results showed that the only determinations of primary (peroxide value) and secondary (TBARs) oxidation products did not allow a sufficient or correct evaluation of the oxidative changes of hemp seed oils during storage. In fact, samples at the end of the test were primarily characterized by a high presence of oxidation volatile compounds and a significant decrease of antioxidants. Several volatiles identified before the accelerated storage, such as the predominant α-pinene and ß-pinene, gradually decreased during the accelerated storage period. On the other hand, aldehydes (hexanal, (E)-2-hexenal, heptanal, (E,E)-2,4-hexadienal, (E)-2-heptenal, (E,E)-2,4-heptadienal, (E,Z)-2,4-heptadienal, 2-octenal, nonanal, nonenal, 2,4-nonadienal, (E,E)- 2,4-decadienal and 2,4-decadienal), ketones (1-octen-3-one, 3-octen-2-one, (E,E)-3,5-octadien-2- one and 3,5-octadien-2-one), acids (propionic acid, pentanoic acid, hexanoic acid and heptanoic acid) and 2-pentyl-furan increased during the accelerated storage, as principal markers of oxidation.

Correction: Endrizzi et al. Relationships between Intensity and Liking for Chemosensory Stimuli in Food Models: A Large-Scale Consumer Segmentation. Foods 2022, 11, 5.

In the original publication [1], there was a mistake in "Figure 2, Figure 3, Figure 4, and Figure 5" as published [...].

Geographical authentication of virgin olive oil by GC-MS sesquiterpene hydrocarbon fingerprint: Verifying EU and single country label-declaration.

According to the last report from the European Union (EU) Food Fraud Network, olive oil tops the list of the most notified products. Current EU regulation states geographical origin as mandatory for virgin olive oils, even though an official analytical method is still lacking. Verifying the compliance of label-declared EU oils should be addressed with the highest priority level. Hence, the present work tackles this issue by developing a classification model (PLS-DA) based on the sesquiterpene hydrocarbon fingerprint of 400 samples obtained by HS-SPME-GC-MS to discriminate between EU and non-EU olive oils, obtaining an 89.6% of correct classification for the external validation (three iterations), with a sensitivity of 0.81 and a specificity of 0.95. Subsequently, multi-class discrimination models for EU and non-EU countries were developed and externally validated (with three different validation sets) with successful results (average of 92.2% of correct classification for EU and 96.0% for non-EU countries).

Food Neophobia, Odor and Taste Sensitivity, and Overall Flavor Perception in Food.

Smell, which allows us to gather information about the hedonic value of an odor, is affected by many factors. This study aimed to assess the relationship among individual factors, odor sensitivity, and enjoyment, and to evaluate how overall flavor perception and liking in actual food samples are affected by odor sensitivity. A total of 749 subjects, from four different Italian regions, participated in the study. The olfactory capabilities test on four odors (anise, banana, mint, and pine), as well as PROP (6-n-prpyl-2-thiouracil) status and food neophobia were assessed. The subjects were clustered into three groups of odor sensitivity, based on the perceived intensity of anise. The liking and intensity of the overall flavor were evaluated for four chocolate puddings with increasing sweetness (C1, C2, C3, and C4). The individual variables significantly affected the perceived intensity and liking of the odors. Even if all of the odor sensitivity groups perceived the more intensely flavored samples as the C1 and C4 chocolate puddings, the high-sensitivity group scored the global flavor of all of the samples as more intense than the low-sensitivity group. The low-sensitive subjects evaluated the liking of the sweeter samples with higher scores than the moderate-sensitive subjects, whereas the high-sensitive subjects gave intermediate scores. In conclusion, odor sensitivity plays a pivotal role in the perception and liking of real food products; this has to be taken into account in the formulation of new products, suitable for particular categories with reduced olfactory abilities.

In-House Validation of an SPE-GC-FID Method for the Detection of Free and Esterified Hydroxylated Minor Compounds in Virgin Olive Oils.

Minor compounds in vegetable oils are distributed between free and esterified forms, and the ratio of these two fractions could represent an important parameter for assessment of oil authenticity. A simple method based on offline SPE-GC-FID for the analysis of free and esterified hydroxylated minor compounds in olive and sunflower oils has been developed and in-house validated. A satisfactory repeatability relative standard deviation (<7.5%) was obtained in all cases. The method, which requires simple instrumentation, allows for reliable quantification in a single chromatographic run with the advantages of minimizing sample manipulation, use of toxic solvents and reagents, and time consumption. The analytical procedure was applied to pure oil samples, including 15 authentic extra virgin olive oils collected from different European countries (Spain, Italy, Greece, and Portugal). Finally, the proposed SPE-GC-FID methodology could detect changes in the ratio between the free and esterified forms in pure extra virgin olive oil when mixed with refined sunflower oil at different percentages of 2, 5, 10, 15, and 20% (w/w) to simulate adulteration.

Relationships between Intensity and Liking for Chemosensory Stimuli in Food Models: A Large-Scale Consumer Segmentation.

This study, which was conducted as part of the Italian Taste project, was aimed at exploring the relationship between actual liking and sensory perception in four food models. Each food model was spiked with four levels of prototypical tastant (i.e., citric acid, sucrose, sodium chloride, capsaicin) to elicit a target sensation (TS) at an increasing perceived intensity. Participants (N = 2258; 59% women, aged 18-60) provided demographic information, a stated liking for 40 different foods/beverages, and their responsiveness to tastants in water. A food-specific Pearson's coefficient was calculated individually to estimate the relationship between actual liking and TS responsiveness. Considering the relationship magnitude, consumers were grouped into four food-specific clusters, depending on whether they showed a strong negative (SNC), a weak negative (WNC), a weak positive (WPC), or a strong positive correlation (SPC). Overall, the degree of liking raised in parallel with sweetness responsiveness, fell as sourness and pungency perception increased, and showed an inverted U-shape relationship with saltiness. The SNC clusters generally perceived TSs at higher intensities, except for sourness. Clusters were validated by associating the level of stated liking towards food/beverages; however, some unexpected indications emerged: adding sugar to coffee or preferring spicy foods differentiated those presenting positive correlations from those showing negative correlations. Our findings constitute a step towards a more comprehensive understanding of food preferences.

Preliminary data on the antimicrobial effect of Cannabis sativa L. variety Futura 75 against food-borne pathogens in vitro as well as against naturally occurring microbial populations on minced meat during storage.

In the present study, the antimicrobial effect of Cannabis sativa Futura 75 was evaluated both in vitro against foodborne bacterial pathogens, and on food against naturally occurring microbial groups of minced meat stored for 8 days at 4°C. Ethanol extraction was performed on the grind of the inflorescence. After extraction, ethanol was completely evaporated and substituted by water. Serial dilutions of the extract, the grind and cannabidiol 99% were added to Nutrient Agar and spotted with Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli and Staphylococcus spp. Regarding the evaluation on food, 50 mL of extract, characterised by CBD at concentration of 322,70 µg/mL, were added to 2.5 kg of minced beef meat. Meat was divided into aliquots and stored for 8 days at 4°C. At 0, 1, 2, 3, 4, 7, and 8 days, aerobic bacteria, enterobacteria, coliforms and E. coli were enumerated. All tested products were efficient against Gram +. In particular, extract corresponding to CBD concentration of 0.017 and 0.3 mg/mL were effective against L. monocytogenes and Staphylococcus spp. respectively. After 8 days of storage at 4°C, treated minced meat showed a bright red colour in comparison to a brownish control meat. Moreover, Enterobacteriaceae and coliforms were significantly reduced of 2.3 log CFU/g and 1.6 log CFU/g respectively in treated meat in comparison to the control. Although preliminary, the present study suggests the antimicrobial properties of the extract of Cannabis sativa both in vitro and in minced meat.

Profiling versus fingerprinting analysis of sesquiterpene hydrocarbons for the geographical authentication of extra virgin olive oils.

The verification of the geographical origin of extra virgin (EVOO) and virgin olive oil (VOO) is crucial to protect consumers from misleading information. Despite the large number of studies performed, specific markers are still not available. The present study aims to evaluate sesquiterpene hydrocarbons (SHs) as markers of EVOO geographical origin and to compare the discrimination efficiency of targeted profiling and fingerprinting approaches. A prospective study was carried out on 82 EVOOs from seven countries, analyzed by Headspace Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS). Classification models were developed by Partial Least Square-Discriminant Analysis (PLS-DA) and internally validated (leave 10%-out cross-validation). The percentage of correct classification was higher for the fingerprinting (100%) than for the profiling approach (45.5-100%). These results confirm the suitability of SHs as EVOO geographical markers and establish the fingerprinting as the most efficient approach for the treatment of SH analytical data with this purpose up to date.

Development and validation of a Fast gas chromatography/mass spectrometry method for the determination of cannabinoids in Cannabis sativa L.

A routine method for determining cannabinoids in Cannabis sativa L. inflorescence, based on Fast gas chromatography coupled to mass spectrometry (Fast GC/MS), was developed and validated. To avoid the decarboxylation of carboxyl group of cannabinoids, different derivatization approaches, i.e. silylation and esterification (diazomethane-mediated), reagents and solvents (pyridine or ethyl acetate), were tested. The methylation significantly increased the signal-to-noise ratio of all carboxylic cannabinoids, except for cannabigerolic acid (CBGA). Since diazomethane is not commercially available, is considered a hazardous reactive and requires 1-day synthesis by specialized chemical staff, silylation was used along the whole validation of a routine method. The method gave a fast (total analysis time < 7.0 min) and satisfactory resolution (R > 1.1), with a good repeatability (intraday < 8.38%; interday < 11.10%) and sensitivity (LOD < 11.20 ng/mL). The Fast GC/MS method suitability for detection of cannabinoids in hemp inflorescences, was tested; a good repeatability (intraday < 9.80%; interday < 8.63%), sensitivity (LOD < 58.89 ng/mg) and robustness (<9.52%) was also obtained. In the analyzed samples, the main cannabinoid was cannabidiolic acid (CBDA, 5.19 ± 0.58 g/100 g), followed by cannabidiol (CBD, 1.56 ± 0.03 g/100 g) and CBGA (0.83 g/100 g). Δ9-tetrahydrocannabivarine (THCV) was present at trace level. Therefore, the developed routine Fast GC/MS method could be a valid alternative for a fast, robust and high sensitive determination of main cannabinoids present in hemp inflorescences.

Detection of Fumonisins in Fresh and Dehydrated Commercial Garlic.

An epidemic fungal disease caused by Fusarium proliferatum, responsible for fumonisin production (FB1, FB2, and FB3), has been reported in the main garlic-producing countries in recent years. Fumonisins are a group of structurally related toxic metabolites produced by this pathogen. The aim of this work was to establish an enzyme-linked immunosorbent assay (ELISA) procedure, mostly applied to cereals, that is suitable for fumonisin detection in garlic and compare these results to those obtained by high-performance liquid chromatography (HPLC) and screening of fresh and dehydrated garlic for toxicological risk. The results show good correlation between the two analytical methods. In fresh symptomatic garlic, fumonisin levels were higher in the basal plates than those in the portions with necrotic spots. Among the 56 commercially dehydrated garlic samples screened, three were positive by ELISA test and only one was above the limit of quantitation. The same samples analyzed by HPLC showed the presence of FB1 in trace amounts that was below the limit of quantitation; FB2 and FB3 were absent. The results are reassuring, because no substantial contamination by fumonisins was found in commercial garlic.

How the Addition of Spices and Herbs to Virgin Olive Oil to Produce Flavored Oils Affects Consumer Acceptance.

With the aim to expand the olive oil market to a larger number of consumers who are not familiar with the sensory characteristics of virgin olive oil, the use of novel products known as "flavored olive oils", obtained by adding different kind of spices and aromatic herbs, is spreading in many countries. In order to test consumer acceptability of this type of product, in a country (Tunisia) in which virgin olive oil is regularly consumed, flavored olive oils were prepared by adding aromatic extracts of thyme, oregano, a mix of herbs (used as pizza seasoning), rosemary, and basil to a monovarietal Chemlali virgin olive oil and a consumer test on 206 subjects was performed. Selected quality parameters (free acidity, peroxide number, oxidative stability, specific absorption at K232 nm and K270 nm) were also measured and no significant variations were detected. Slight differences were found concerning the content of minor compounds (chlorophylls, carotenoids and total phenols). On the other hand, notable differences were seen in the profiles of volatile compounds, which appeared to be responsible for the observed variability in consumer acceptance. Although the unflavored oil was more appreciated than the flavored ones, among the latter, thyme flavored olive oil was the most appreciated.

Agricultural by-products with bioactive effects: A multivariate approach to evaluate microbial and physicochemical changes in a fresh pork sausage enriched with phenolic compounds from olive vegetation water.

The use of phenolic compounds derived from agricultural by-products could be considered as an eco-friendly strategy for food preservation. In this study a purified phenol extract from olive vegetation water (PEOVW) was explored as a potential bioactive ingredient for meat products using Italian fresh sausage as food model. The research was developed in two steps: first, an in vitro delineation of the extract antimicrobial activities was performed, then, the PEOVW was tested in the food model to investigate the possible application in food manufacturing. The in vitro tests showed that PEOVW clearly inhibits the growth of food-borne pathogens such as Listeria monocytogenes and Staphylococcus aureus. The major part of Gram-positive strains was inhibited at the low concentrations (0.375-3mg/mL). In the production of raw sausages, two concentrates of PEOVW (L1: 0.075% and L2: 0.15%) were used taking into account both organoleptic traits and the bactericidal effects. A multivariate statistical approach allowed the definition of the microbial and physicochemical changes of sausages during the shelf life (14days). In general, the inclusion of the L2 concentration reduced the growth of several microbial targets, especially Staphylococcus spp. and LABs (2log10CFU/g reduction), while the increasing the growth of yeasts was observed. The reduction of microbial growth could be involved in the reduced lipolysis of raw sausages supplemented with PEOVW as highlighted by the lower amount of diacylglycerols. Moisture and aw had a significant effect on the variability of microbiological features, while food matrix (the sausages' environment) can mask the effects of PEOVW on other targets (e.g. Pseudomonas). Moreover, the molecular identification of the main representative taxa collected during the experimentation allowed the evaluation of the effects of phenols on the selection of bacteria. Genetic data suggested a possible strain selection based on storage time and the addition of phenol compounds especially on LABs and Staphylococcus spp. The modulation effects on lipolysis and the reduction of several microbial targets in a naturally contaminated product indicates that PEOVW may be useful as an ingredient in fresh sausages for improving food safety and quality.

Cardiac and Vascular Synergic Protective Effect of Olea europea L. Leaves and Hibiscus sabdariffa L. Flower Extracts.

This study was aimed at investigating the cardiovascular effects of an Olea europea L. leaf extract (OEE), of a Hibiscus sabdariffa L. flower extract (HSE), and of their 13 : 2 w/w mixture in order to assess their cardiac and vascular activity. Both extracts were fully characterized in their bioactive compounds by HPLC-MS/MS analysis. The study was performed using primary vascular endothelial cells (HUVECs) to investigate the antioxidant and cytoprotective effect of the extracts and their mixture and isolated guinea-pig left and right atria and aorta to evaluate the inotropic and chronotropic activities and vasorelaxant properties. In cultured HUVECs, OEE and HSE reduced intracellular reactive oxygen species formation and improved cell viability, following oxidative stress in dose-dependent manner. OEE and HSE exerted negative inotropic and vasorelaxant effects without any chronotropic property. Interestingly, the mixture exerted higher cytoprotective effects and antioxidant activities. Moreover, the mixture exerted an inotropic effect similar to each single extract, while it revealed an intrinsic negative chronotropic activity different from the single extract; its relaxant activity was higher than that of each single extract. In conclusion OEE and HSE mixture has a good potential for pharmaceutical and nutraceutical application, thanks to the synergistic effects of the single phytochemicals.

Liquid-liquid and solid-phase extractions of phenols from virgin olive oil and their separation by chromatographic and electrophoretic methods.

The high oxidative stability of virgin olive oil is related to its high monounsaturated/polyunsaturated ratio and to the presence of antioxidant compounds, such as tocopherols and phenols. In this paper, the isolation of phenolic compounds from virgin olive oil, by different methods, was tested and discussed. Particularly liquid-liquid and solid-phase extraction methods were compared, assaying, for the latter, three stationary phases (C8, C18 and Diol) and several elution mixtures. Quantification of phenolic and o-diphenolic substances in the extracts was performed by the traditional Folin-Ciocalteau method and the sodium molybdate reaction, respectively. Furthermore, the quantification of phenolic compounds in the extracts and in a standard mixture was carried out both with diode array and mass spectrometric detection and capillary zone electrophoresis.

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.