Optimizing the Integration of Microwave Processing and Enzymatic Extraction to Produce Polyphenol-Rich Extracts from Olive Pomace.

The integration of green technologies such as microwave- and enzyme-assisted extraction (MEAE) has been shown to improve the extraction efficiency of bioactive compounds while reducing processing time and costs. MEAE using tannase alone (MEAE-Tan), or in combination with cellulase and pectinase (MEAE-Tan-Cel-Pec), was optimized to produce enriched phenolic and antioxidant extracts from olive pomace. The individual and integrated impact of enzyme concentration, temperature, and pomace/water ratio were determined using a central composite rotatable design. Optimal extraction conditions for MEAE-Tan (60 °C, 15 min, 2.34% of enzyme (w/w), and 1:15 pomace/water ratio) and MEAE-Tan-Cel-Pec (46 °C, 15 min, 2% of enzymes (w/w), in the proportion of 1:1:1, and 1:20 pomace/water ratio) resulted in extracts containing 7110.6 and 2938.25 mg GAE/kg, respectively. The antioxidant activity of the extracts was correlated with phenolic acid release, which was enzyme-dependent, as determined with HPLC-DAD analysis. Enzyme selection had a significant impact on the phenolic profile of extracts, with tannase releasing high concentrations of chlorogenic acid and the combined use of enzymes releasing high concentrations of hydroxytyrosol and chlorogenic and ferulic acids. The novelty of this study relies on the integration and optimization of two green technologies (microwave- and enzyme-assisted extraction) to improve the extraction efficiency of bioactive phenolics from olive pomace while reducing processing time and costs. While these techniques have been evaluated isolated, the benefits of using both processing strategies simultaneously remain largely unexplored. This study demonstrates the effectiveness of the integration and processing optimization of two environmentally friendly technologies as a promising alternative to treat agro-industrial byproducts.

Oxidative Stability of Walnut Kernel and Oil: Chemical Compositions and Sensory Aroma Compounds.

The impact of storage temperature and time on quality of two walnut cultivars (Juglans regia Chandler and Howard) were evaluated. Free fatty acids, peroxides, and oxidative stabilities exhibited significant changes. After the storage period, γ-, δ-, and α-tocopherols in Howard oil significantly reduced by 42, 56, and 100% at 5 °C, while 23 °C showed 48, 42, and 100% losses, respectively. For Chandler oil, storage at 5 °C reduced γ-, δ-, and α-tocopherols by 19, 24, and 100%, while 23 °C caused 42, 45, and 100% losses, respectively. Storage of Howard kernels, up to month four, significantly reduced total phenolics by 9 and 18%, at 23 and 5 °C, respectively, whereas Chandler also reduced by 9 and 27%, at 23 and 5 °C, respectively. Additionally, 14 phenolic compounds were profiled in kernels, where flavonoids were dominant than phenolic acids. At the end of month four, the dominant phenolic compound was gallic acid at 23 °C (981.68 and 703 mg/kg for Chandler and Howard, respectively). Additionally, positive correlations were observed between rancid sensory perceptions and oxidative volatiles. Storage conditions are crucial for maintaining the sensory and nutritional attributes of walnuts during postharvest management.

Extraction, Purification and In Vitro Antioxidant Activity Evaluation of Phenolic Compounds in California Olive Pomace.

Olive pomace (OP) is a valuable food byproduct that contains natural phenolic compounds with health benefits related to their antioxidant activities. Few investigations have been conducted on OP from the United States while many studies on European OP have been reported. OP of Arbequina, the most common cultivar from California, was collected and extracted by water, 70% methanol and 70% ethanol, followed by purification using macroporous absorbing resin. Results showed that the extractable total phenolic content (TPC) was 36-43 mg gallic acid equivalents (GAE)/g in pitted, drum-dried defatted olive pomace (DOP), with major contributions from hydroxytyrosol, oleuropein, rutin, verbascoside, 4-hydroxyphenyl acetic acid, hydroxytyrosol-glucoside and tyrosol-glucoside. Macroporous resin purification increased TPC by 4.6 times the ethanol crude extracts of DOP, while removing 37.33% total sugar. The antioxidant activities increased 3.7 times Trolox equivalents (TrE) by DPPH and 4.7 times TrE by ferric reducing antioxidant power (FRAP) in the resin purified extracts compared to the ethanol crude extracts. This study provided a new understanding of the extraction of the bioactive compounds from OP which could lead to practical applications as natural antioxidants, preservatives and antimicrobials in clean-label foods in the US.

Geographical Origin Assessment of Extra Virgin Olive Oil via NMR and MS Combined with Chemometrics as Analytical Approaches.

Geographical origin assessment of extra virgin olive oil (EVOO) is recognised worldwide as raising consumers' awareness of product authenticity and the need to protect top-quality products. The need for geographical origin assessment is also related to mandatory legislation and/or the obligations of true labelling in some countries. Nevertheless, official methods for such specific authentication of EVOOs are still missing. Among the analytical techniques useful for certification of geographical origin, nuclear magnetic resonance (NMR) and mass spectroscopy (MS), combined with chemometrics, have been widely used. This review considers published works describing the use of these analytical methods, supported by statistical protocols such as multivariate analysis (MVA), for EVOO origin assessment. The research has shown that some specific countries, generally corresponding to the main worldwide producers, are more interested than others in origin assessment and certification. Some specific producers such as Italian EVOO producers may have been focused on this area because of consumers' interest and/or intrinsic economical value, as testified also by the national concern on the topic. Both NMR- and MS-based approaches represent a mature field where a general validation method for EVOOs geographic origin assessment could be established as a reference recognised procedure.

Low Acrylamide Flatbreads Prepared from Colored Rice Flours and Relationship to Asparagine and Proximate Content of Flours and Flatbreads.

Acrylamide is a potentially toxic compound present in many plant-based foods, such as coffee, breads, and potato fries, which is reported to have carcinogenic, neurotoxic, and antifertility properties in vivo, suggesting the need to keep the acrylamide content of widely consumed food as low as possible. As pigmented rice contains bioactive phenolic and flavonoid compounds, the objective of this study was to potentially enhance the beneficial properties of flatbreads by evaluating the acrylamide content and proximate composition of 12 novel flatbreads prepared from the following commercial pigmented rice seeds: Black Japonica, Chinese Black, French Camargue, Himalayan Red, Long Grain Brown, Purple Sticky, Short Grain Brown, Wehani, Wild, Indian Brown Basmati, Organic Brown Jasmine, and Organic Jade Pearl. Although acrylamide levels ranged from 4.9 µg/kg in Long Grain Brown to 50.8 µg/kg in Chinese Black, the absolute values were all low (though statistically significantly differences existed among varieties). Acrylamide content did not correlate with its precursor asparagine. The variations in protein, carbohydrate, fat, ash, dry matter, and water content determined by proximate analysis, and the reported health benefits of colored rice cultivars used to prepare the flatbreads, might also be useful for relating composition to nutritional qualities and health properties, facilitating their use as nutritional and health-promoting functional foods.

Low Acrylamide Flatbreads from Colored Corn and Other Flours.

Dietary acrylamide formed during baking and frying of plant-based foods such as bread and other cereal products, coffee, fried potatoes, and olives is reported to induce genotoxic, carcinogenic, neurotoxic, and antifertility properties in vivo, suggesting the need to keep the acrylamide content low with respect to widely consumed heat-processed food including flatbreads. Due to the fact that pigmented corn flours contain biologically active and health-promoting phenolic and anthocyanin compounds, the objective of this study was to potentially define beneficial properties of flatbread by evaluating the acrylamide content determined by high-performance liquid chromatography/mass spectrometry (HPLC/MS) with a detection limit of 1.8 µg/kg and proximate composition by standard methods of six experimental flatbreads made from two white, two blue, one red, and one yellow corn flours obtained by milling commercial seeds. Acrylamide content was also determined in experimental flatbreads made from combinations in quinoa flour, wheat flour, and peanut meal with added broccoli or beet vegetables and of commercial flatbreads including tortillas and wraps. Proximate analysis of flatbreads showed significant differences in protein and fat but not in carbohydrate, mineral, and water content. The acrylamide content of 16 evaluated flatbreads ranged from 0 to 49.1 µg/kg, suggesting that these flatbreads have the potential to serve as low-acrylamide functional foods. The dietary significance of the results is discussed.

Walnut (Juglans regia L.) Volatile Compounds Indicate Kernel and Oil Oxidation.

Kernel oxidation susceptibility and pellicle darkening are among the biggest concerns regarding walnut quality. Monitoring oxidation is crucial to preserve quality from production to consumption. Chemical oxidation parameters (peroxide value and UV absorbances), fatty acid profile, tocopherols, phenols, and volatiles in 'Chandler' and 'Howard' kernels were studied at different time points during 28 weeks of storage to evaluate potential oxidation markers. During storage, peroxide value, UV absorbances, and volatiles concentration increased; oxidative stability, phenols, and tocopherols decreased, while fatty acid profile was unaffected. 'Chandler' had a lower peroxide value, K232, and K268; and higher kernel and oil oxidative stability compared to 'Howard'. Phenols and tocopherols decreased 1.2-fold in 'Chandler' and 1.3-fold in 'Howard'. Using multivariate analysis, samples were discriminated in three groups according with their oxidative levels. Increases of volatiles in oil and kernel were associated with higher oxidative levels. Pentanal, 2-methylpropanal, hexanal, (E)-2-pentenal, 3-octanone, octanal, (Z)-2-penten-1-ol, hexanol, (E)-2-octenal, 1-octen-3-ol, benzaldehyde, (E,E)-2,4-nonadienal, and hexanoic acid in kernels were adequate at distinguishing oxidation levels and as oxidative markers in walnuts. Kernel volatiles is a useful measurement for walnut oxidation during storage without any prior fat extraction.

Analysis and Authentication of Avocado Oil Using High Resolution NMR Spectroscopy.

Avocado oil is a food product of high commercial and nutritional value. As a result, it can be a subject of adulteration similar to other high-value edible oils, such as olive oil. For olive oil and many other foods products, NMR spectroscopy has been successfully used for authentication and quality assessment. In this study, we apply NMR analysis to avocado oil to differentiate it from other oils including olive, canola, high-oleic (HO) safflower, HO sunflower and soybean oil using commercial and lab-made samples of avocado oils. NMR allowed the rapid analysis of the fatty acid profile and detection of minor compounds, such as sterols, oxidation products, and hydrolysis products, which can be used to assess oil quality and authenticity. The NMR assignment was conducted using traditional 2D NMR and the novel NOAH super-sequences. Combining chemometrics with NMR enabled us to differentiate between avocado oil and other oils. Avocado oil has compositional similarities with other vegetable oils, such as HO sunflower and HO safflower oil, which can be used as potential adulterants. Despite these similarities, NMR-based metabolomics captured differences in the levels of certain compounds including fatty acids, terpenes, sterols, and oxidation products to detect adulteration and for quality control purposes.

Comparative Effect of Hammer Mill Screen Size and Cell Wall-Degrading Enzymes During Olive Oil Extraction.

The influence of hammer mill screen size (4.5 and 8.5 mm) and enzyme addition (control and 500 ppm) on olive fruit cell wall breakdown and its consequences in terms of oil recovery and the phenolic content of olive oil was studied at the laboratory scale for "Arbequina" and "Koroneiki" at two different maturities. Water recovery and water-soluble carbohydrates in olive paste after malaxation were measured as an indicator of cell wall breakdown. Smaller screen size and enzymes increase oil recovery for Arbequina with a maturity index of 1.6 (6.3-6.6%); and for Koroneiki at a maturity index of 0.2 (15.0-38%) and 2.6 (1.3-4.3%). For both cultivars, the increase in oil recovery is larger in green fruits compared to more ripe fruit. Water recovery and water-soluble carbohydrates increase with small screen size and the enzyme treatments, even when no increment in oil recovery is observed. The water recovery range was 143-239% for Arbequina and 150-262% for Koroneiki; water-soluble carbohydrate range was 1.8-12.7 g/kg for Arbequina and 0.5-5.4 g/kg for Koroneiki. In general, smaller hammer mill screen size and enzymes increase total phenols in the oil, with a larger difference between control and treatment for green fruit than for the ripe fruit. For Arbequina, increases in total phenol content were in the range of 45-60 and 5-20% at maturity index 1.6 and 3.3, respectively. For Koroneiki, the increases were in the range of 31-121 and 7-9% at maturity index 0.2 and 2.6, respectively. Application of cell wall-degrading enzymes improves the cell wall breakdown caused by hammer mill, leading to higher oil recovery and total phenol content. The magnitude of the effect depends on the cultivar and olive fruit maturity.

Study of the minor fraction of virgin olive oil by a multi-class GC-MS approach: Comprehensive quantitative characterization and varietal discrimination potential.

For the first time, a multi-class GC-MS method was applied to perform the quantitative-profiling of the minor fraction of VOOs (considering >40 compounds) in a single run. This comprehensive methodology has demonstrated a remarkable profiling ability on five groups of compounds (phenolic and triterpenic compounds, tocopherols, sterols and free fatty acids) with wide range of polarities/volatilities and chemical entities. After the complete analytical validation of the method, 32 VOO samples from eight different cultivars (some of them very scarcely studied before) were analyzed and the quantitative results were subjected to both non-supervised and supervised multivariate statistics for testing the capability of the determined VOO minor compounds to discriminate the varietal origin of the samples. Typical compositional profiles were defined for each cultivar and promising potential varietal markers were pointed out. The models built to discriminate Cayon and Maurino samples from the rest exhibited the best quality parameters. The relative levels of tocopherols together with characteristic concentration of luteolin, ß-sitosterol and tyrosol were, for instance, the most specific features of Cayon VOOs.

Influence of fruit canopy position and maturity on yield determinants and chemical composition of virgin olive oil.

BACKGROUND: While both maturity and light exposure are important factors determining olive fruit physiology, the relationship between maturity, canopy position and optimal harvesting time has not been well-studied. To understand the interaction of these factors, olive fruits from upper and lower layers of the canopy were harvested from September to January. Maturity, moisture and fat content of the fruit as well as the quality and minor components of the oil extracted were measured. RESULTS: Lower light interception at the lower canopy positions resulted in differences in the fruits and oil extracted between canopy layers. Upper layer presented 60% of the overall production; fruit had one unit more of maturity index, 3% less moisture and 5% more fat content. Oil extracted from the upper layers presented higher concentration of oleuropein and ligstroside aglycone. Fruits from upper layers at maturity index of two had higher fat content and more total phenols in the oil extracted when compared with fruits from lower layer with the same maturity index. CONCLUSIONS: Differences in oil composition between layers do not correlate with differences in the fruit maturity index; instead, fruit position is a determining factor for physiological processes related to fruit growth and oil composition. © 2019 Society of Chemical Industry.

Phenolics and Antioxidant Capacity of Pitted Olive Pomace Affected by Three Drying Technologies.

This study investigated drum-drying's ability to produce dried food-grade olive pomace as a potential food ingredient that is more nutritionally dense than its freeze-dried and hot-air dried counterparts. The pits and skin were removed from fresh olive pomace, and the remaining pulp was dried to <5% moisture through freeze-drying, hot-air drying, and drum-drying at two rotational speeds. The drying treatments had no significant (P ≤ 0.05) effect on the olive pomace's fat or dietary fiber contents but did increase the L* , a* , and b* color parameter values. Although all the drying treatments significantly (P ≤ 0.05) decreased the fresh olive pomace's antioxidant capacity, drum-drying preserved the olive pomace's antioxidant capacity significantly (P ≤ 0.05) better than freeze-drying and hot-air drying. The drum-dried samples had concentrations of caffeic acid and verbascoside that were significantly (P ≤ 0.05) higher than the other dried pomace samples and were not significantly (P ≤ 0.05) different from the fresh pomace. The drum-dried olive pomace contained concentrations of hydroxytyrosol, tyrosol, vanillic acid, luteolin-7-glucoside, and rutin that were not significantly (P ≤ 0.05) different from the dried sample with the highest concentration of each respective phenolic compound. No oleuropein was found in the fresh or dried olive pomace. The results of this study show that drum-drying is an energy efficient method for converting olive pomace into a stable food-grade supplement that preserves its high phenolic, antioxidant, and dietary fiber contents to potentially benefit human health when incorporated into food or supplement products. PRACTICAL APPLICATION: Pitting and drying converts the olive pomace into a stable form that is free of physical hazards and could be incorporated into food products to increase their nutritional quality through olive pomaces' high fiber, antioxidant, and phenolic contents. Drum-drying allows food-grade olive pomace to retain higher amounts of beneficial soluble phenolics and a higher antioxidant capacity than conventional drying methods, thus furthering olive pomace's potential valorization as a food ingredient.

Deep insight into the minor fraction of virgin olive oil by using LC-MS and GC-MS multi-class methodologies.

Several analytical methods are available to evaluate virgin olive oil (VOO) minor compounds; however, multi-class methodologies are yet rarely studied. Herewith, LC-MS and GC-MS platforms were used to develop two methods capable of simultaneously determine more than 40 compounds belonging to different VOO minor chemical classes within a single run. A non-selective and highly efficient liquid-liquid extraction protocol was optimized for VOO minor components isolation. The separation and detection conditions were adjusted for determining phenolic and triterpenic compounds, free fatty acids and tocopherols by LC-MS, plus sterols and hydrocarbons by GC-MS. Chromatographic analysis times were 31 and 50 min, respectively. A comparative assessment of both methods in terms of analytical performance, easiness, cost and adequacy to the analysis of each class was carried out. The emergence of this kind of multi-class analytical methodologies greatly increases throughput and reduces cost, while avoiding the complexity and redundancy of single-chemical class determinations.

Impact of industrial hammer mill rotor speed on extraction efficiency and quality of extra virgin olive oil.

Crushing is a key step during olive oil extraction. Among commercial crushers, the hammer mill is the most widely used due to its robustness and high throughput. In the present work, the impact of hammer mill rotor speed on extraction yield and overall quality of super-high-density Arbosana olive oils were assessed in an industrial facility. Our results show that increasing the rotor speed from 2400rpm to 3600rpm led to a rise in oil yield of 1.2%, while conserving quality parameters. Sensory analysis showed more pungency with increased rotation speed, while others attributes were unaffected. Volatile compounds showed little variation with the differences in crusher speed; however, total phenols content, two relevant secoiridoids, and triterpenoids levels increased with rotor speed. Hammer mill rotor speed is a processing variable that can be tuned to increase the extraction efficiency and modulate the chemical composition of extra virgin olive oil.

Oleocanthal-rich extra virgin olive oil demonstrates acute anti-platelet effects in healthy men in a randomized trial.

The phenolic profiles of extra virgin olive oils (EVOOs) may influence their cardiovascular benefits. In a randomized crossover of acute EVOO intake on platelet function, participants (n=9) consumed 40 mL of EVOO weekly. EVOOs were matched for total phenolic content and were either tyrosol-poor with 1:2 oleacein/oleocanthal (D2i0.5), or 2:1 oleacein/oleocanthal (D2i2), or predominantly tyrosol (D2i0). Ibuprofen provided a platelet inhibition control. Blood was collected pre- and 2 hr post-EVOO intake. D2i0.5 and D2i2 reduced 1 µg/mL collagen-stimulated maximum platelet aggregation (Pmax), with effects best correlated to oleocanthal intake (R=0.56, P=0.002). Total phenolic intake was independently correlated to eicosanoid production inhibition, suggesting that cyclooxygenase blockade was not responsible for the Pmax inhibition. Five participants exhibited >25% ΔPmax declines with D2i0.5 and D2i2 intake and plasma metabolomic profiles discriminated subjects by oil responsivity. Platelet responses to acute EVOO intake are associated with oil phenolic composition and may be influenced by diet.

High-performance liquid chromatography with fluorescence detection for the rapid analysis of pheophytins and pyropheophytins in virgin olive oil.

Pheophytins and pyropheophytin are degradation products of chlorophyll pigments, and their ratios can be used as a sensitive indicator of stress during the manufacturing and storage of olive oil. They increase over time depending on the storage condition and if the oil is exposed to heat treatments during the refining process. The traditional analysis method includes solvent- and time-consuming steps of solid-phase extraction followed by analysis by high-performance liquid chromatography with ultraviolet detection. We developed an improved dilute/fluorescence method where multi-step sample preparation was replaced by a simple isopropanol dilution before the high-performance liquid chromatography injection. A quaternary solvent gradient method was used to include a fourth strong solvent wash on a quaternary gradient pump, which avoided the need to premix any solvents and greatly reduced the oil residues on the column from previous analysis. This new method not only reduces analysis cost and time but shows reliability, repeatability, and improved sensitivity, especially important for low-level samples.

In-depth assessment of analytical methods for olive oil purity, safety, and quality characterization.

This paper evaluates the performance of the current analytical methods (standard and widely used otherwise) that are used in olive oil for determining fatty acids, triacylglycerols, mono- and diacylglycerols, waxes, sterols, alkyl esters, erythrodiol and uvaol, tocopherols, pigments, volatiles, and phenols. Other indices that are commonly used, such as free acidity and peroxide value, are also discussed in relation to their actual utility in assessing quality and safety and their possible alternatives. The methods have been grouped on the basis of their applications: (i) purity and authenticity; (ii) sensory quality control; and (iii) unifying methods for different applications. The speed of the analysis, advantages and disadvantages, and multiple quality parameters are assessed. Sample pretreatment, physicochemical and data analysis, and evaluation of the results have been taken into consideration. Solutions based on new chromatographic methods or spectroscopic analysis and their analytical characteristics are also presented.

Quantum chemical study of the isomerization of 24-methylenecycloartanol, a potential marker of olive oil refining.

Quantum chemical calculations on the isomerization of 24-methylenecycloartanol are described. An energetically viable mechanism, with a rate-determining protonation step, is proposed. This rearrangement may find applicability in tests for determining if an olive oil has been refined.

Biomimetic total synthesis of santalin†Y.

A biomimetic total synthesis of santalin Y, a structurally complex but racemic natural product, is described. The key step is proposed to be a (3+2) cycloaddition of a benzylstyrene to a "vinylogous oxidopyrylium", which is followed by an intramolecular Friedel-Crafts reaction. This cascade generates the unique oxafenestrane framework of the target molecule and sets its five stereocenters in one operation. Our work provides rapid access to santalin Y and clarifies its biosynthetic relationship with other colorants isolated from red sandalwood.

Linking chemical parameters to sensory panel results through neural networks to distinguish olive oil quality.

A wide variety of olive oil samples from different origins and olive types has been chemically analyzed as well as evaluated by trained sensory panelists. Six chemical parameters have been obtained for each sample (free fatty acids, peroxide value, two UV absorption parameters (K232 and K268), 1,2-diacylglycerol content, and pyropheophytins) and linked to their quality using an artificial neural network-based model. Herein, the nonlinear algorithms were used to distinguish olive oil quality. Two different methods were defined to assess the statistical performance of the model (a K-fold cross-validation (K = 6) and three different blind tests), and both of them showed around a 95-96% correct classification rate. These results support that a relationship between the chemical and the sensory analyses exists and that the mathematical tool can potentially be implemented into a device that could be employed for various useful applications.