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.

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.

Biosynthesis and Conformational Properties of the Irregular Sesquiterpenoids Isothapsadiene and ß-Isothapsenol.

A carbocation cyclization/rearrangement mechanism for the biosynthesis of isothapsadiene and ß-isothapsenol is shown to be energetically viable on the basis of density functional theory (DFT) calculations. In addition, for both isothapsadiene and ß-isothapsenol, variable-temperature NMR experiments reveal two equilibrium conformers that undergo hindered exchange. The identities of these conformers, which are related by a chair-flip, are confirmed by DFT calculations on their structures, energies, 1H and 13C chemical shifts, and interconversion pathways.

Viability of dodecahedrane-forming radical polycyclizations.

The results of density functional theory calculations on thiyl radical-promoted polycyclization to form dodecahedrane are described. On the basis of these results, the proposed polycyclization is predicted to be both highly exergonic and fast. These results showcase the potential power of radical polycyclization reactions in constructing complex molecular architectures.

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.

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.

Physical and chemical characteristics of walnut (Juglans regia L.) kernels with different skin lightness.

Walnuts undergo rigorous grading before being sold to customers. There are multiple parameters used for the grading, including skin lightness. Walnuts with light skin receive superior grades while walnuts with dark skin are given poor grades or even rejected. However, information on the quality and physicochemical properties of walnuts with varying skin lightness levels is minimal. Therefore, we studied the quality of kernels of varying skin lightness from three common cultivars grown in California, USA (Chandler, Howard, and Tulare). The samples were subjected to size and weight, fat content, free fatty acid, peroxide value, oxidative stability, volatiles, tocopherols, fatty acid profile, and phenol measurements. The dark kernels had significantly lower weight and fat content, higher oxidative stability, and more volatiles than their light counterparts. The dark kernels had higher concentrations of some phenolics but low procyanidin B1 and non-existent epicatechin gallate, compared to the light kernels, indicating that these two phenolics were likely involved in an antioxidant mechanism. Oxidation and depletion of epicatechin gallate likely contributed to the darkening of walnut color.

Analysis and authentication of avocado oil by low-field benchtop NMR spectroscopy and chemometrics.

Avocado oil is a nutritious, edible oil produced from avocado fruit. It has high commercial value and is increasing in popularity, thus powerful analytical methods are needed to ensure its quality and authenticity. Recent advancements in low-field (LF) NMR spectroscopy allow for collection of high-quality data despite the use of low magnetic fields produced by non-superconductive magnets. Combined with chemometrics, LF NMR opens new opportunities in food analysis using targeted and untargeted approaches. Here, it was used to determine poly-, mono-, and saturated fatty acids in avocado oil. Although direct signal integration of LF NMR spectra was able to determine certain classes of fatty acids, it had several challenges arising from signal overlapping. Thus, we used partial least square regression and developed models with good prediction performance for fatty acid composition, with residual prediction deviation ranging 3.46-5.53 and root mean squared error of prediction CV ranging 0.46-2.48. In addition, LF NMR, combined with unsupervised and supervised methods, enabled the differentiation of avocado oil from other oils, namely, olive oil, soybean oil, canola oil, high oleic (OL) safflower oil, and high OL sunflower oil. This study showed that LF NMR can be used as an efficient alternative for the compositional analysis and authentication of avocado oil. PRACTICAL APPLICATION: Here, we describe the application of LF-NMR for fatty acid analysis and avocado oil authentication. LF-NMR can be an efficient tool for targeted and untargeted analysis, thus becoming an attractive option for companies, regulatory agencies, and quality control laboratories. This tool is especially important for organizations and entities seeking economic, user-friendly, and sustainable analysis solutions.

Analysis of mousy off-flavor compound 2-Acetyl-tetrahydropyridine using Liquid Chromatography Mass Spectrometry with Electrospray Ionization in sour beer.

Mousy off-flavor describes N-heterocycles compounds related to spoilage in the brewing industry. It has also been identified in sour beers through sensory analysis. Therefore, preventing spoilage N-heterocycles development is essential to preserve end-products and obviate economic losses. To this day, no methods or protocols have been reported to identifying mousy off-flavor compounds in a beer matrix. The main objective of this work was to develop a standardized quantification method for 2-acetyl-3,4,5,6-tetrahydropyridine (ATHP) in beer matrix, by Liquid Chromatography Mass Spectrometry with Electrospray Ionization (LC-MS-ESI). Extraction of ATHP in the samples was performed using QuEChERS (quick, easy, cheap, effective, rugged, and safe) technique. Over a dozen different potentially mousy cask-aged sour beers including other spontaneously fermented beverages were provided, based on sensory analysis, to determine the variation in ATHP levels. Results indicated ATHP was found in all the samples, ranging from 1.64 ± 0.06 to 57.96 ± 2.15 µg L-1. Herein, we described our detection method of mousy-off flavor compounds which enables future research to mitigate the occurrence of such defects in fermented beverages matrix.•ATHP content in samples varied from 1.64 ± 0.06 to 57.96 ± 2.15 µg L-1.•The recovery range of ATHP using LC-MS-ESI varied from 71% to 97%.•Basified QuEChERS salting-out procedure is applicable for ATHP extraction from beer and other fermented beverages matrices.

A Review of N-Heterocycles: Mousy Off-Flavor in Sour Beer.

Beer has over 600 flavor compounds and creates a positive tasting experience with acceptable sensory properties, which are essential for the best consumer experience. Spontaneous and mixed-culture fermentation beers, generally classified as sour beers, are gaining popularity compared to typical lager or ale styles, which have dominated in the USA for the last few decades. Unique and acceptable flavor compounds characterize sour beers, but some unfavorable aspects appear in conjunction. One such unfavorable flavor is called "mousy". This description is usually labeled as an unpleasant odor, identifying spoilage of fermented food and beverages. It is related as having the odor of mouse urine, cereal, corn tortilla chips, or freshly baked sour bread. The main compounds responsible for it are N-heterocyclic compounds: 2-acetyltetrahydropyridine, 2-acetyl-1-pyrroline, and 2-ethyltetrahydropyridine. The most common beverages associated with mousy off-flavor are identified in wines, sour beers, other grain-based beverages, and kombucha, which may contain heterofermentative lactic acid bacteria, acetic acid bacteria, and/or yeast/fungus cultures. In particular, the fungal species Brettanomyces bruxellensis are associated with mousy-off flavor occurrence in fermented beverages matrices. However, many factors for N-heterocycle formation are not well-understood. Currently, the research and development of mixed-cultured beer and non/low alcohol beverages (NABLAB) has increased to obtain the highest quality, sensory, functionality, and most notably safety standards, and also to meet consumers' demand for a balanced sourness in these beverages. This paper introduces mousy off-flavor expression in beers and beverages, which occurs in spontaneous or mixed-culture fermentations, with a focus on sour beers due to common inconsistency aspects in fermentation. We discuss and suggest possible pathways of mousy off-flavor development in the beer matrix, which also apply to other fermented beverages, including non/low alcohol drinks, e.g., kombucha and low/nonalcohol beers. Some precautions and modifications may prevent the occurrence of these off-flavor compounds in the beverage matrix: improving raw material quality, adjusting brewing processes, and using specific strains of yeast and bacteria that are less likely to produce the off-flavor. Conceivably, it is clear that spontaneous and mixed culture fermentation is gaining popularity in industrial, craft, and home brewing. The review discusses important elements to identify and understand metabolic pathways, following the prevention of spoilage targeted to off-flavor compounds development in beers and NABLABs.

Theoretical and experimental analysis of the reaction mechanism of MrTPS2, a triquinane-forming sesquiterpene synthase from chamomile.

Terpene synthases, as key enzymes of terpene biosynthesis, have garnered the attention of chemists and biologists for many years. Their carbocationic reaction mechanisms are responsible for the huge variety of terpene structures in nature. These mechanisms are amenable to study by using classical biochemical approaches as well as computational analysis, and in this study we combine quantum-chemical calculations and deuterium-labeling experiments to elucidate the reaction mechanism of a triquinane forming sesquiterpene synthase from chamomile. Our results suggest that the reaction from farnesyl diphosphate to triquinanes proceeds through caryophyllyl and presilphiperfolanyl cations and involves the protonation of a stable (-)-(E)-ß-caryophyllene intermediate. A tyrosine residue was identified that appears to be involved in the proton-transfer process.

Upcycling flavanol-rich Chardonnay and Pinot noir grape thinned clusters as potentially functional food ingredients in cocoa-based products.

In California, over 3.4 million tons of wine grapes were crushed in 2020 while every year roughly 20% of the grape mass goes unused. Grape cluster thinning at veraison, a common agricultural practice to ensure color homogeneity in wine grapes, adds to the production costs and generates substantial on-farm loss during grapevine cultivation in which the health-promoting values of thinned clusters (unripe grapes) are usually overlooked. In particular, the health-promoting properties of flavanol monomers, specifically (+)-catechin and (-)-epicatechin, and their oligomeric procyanidins, have been extensively studied in cocoa and chocolate but not so much in grape thinned clusters in recent epidemiology studies. As part of the important agricultural by-products upcycling effort, the current study compared thinned clusters from Chardonnay and Pinot noir, two premium wine grape varieties cultivated in California, to a traditionally Dutch (alkalized) cocoa powder that has been widely used in food applications. Thinned cluster fractions from Chardonnay and Pinot noir grapes grown in the North Coast of California showed much higher concentrations of flavanol monomers and procyanidins, with 208.8-763.5 times more (+)-catechin, 3.4-19.4 times more (-)-epicatechin, and 3.8-12.3 times more procyanidins (by degree of polymerization DP 1-7) than those in the traditionally Dutch cocoa powder. These flavanol-rich thinned clusters that are also considered as plant-based natural products suggested great potential to be functional ingredients in cocoa-based products-which have been ubiquitously perceived as flavanol-rich products by consumers-to enhance their overall dietary flavanol content.

Bulk and Compound-Specific Stable Isotope Analysis for the Authentication of Walnuts (Juglans regia) Origins.

Walnuts are grown in various countries, and as product origin information is becoming more important to consumers, new techniques to differentiate walnut geographical authenticity are needed. We conducted bulk stable isotope analysis (BSIA) and compound-specific stable isotope analysis (CSIA) on walnuts grown in seven countries. The BSIA consisted of δ13Cbulk, δ15Nbulk, and δ34Sbulk, and CSIA covered δ2Hfatty acid, δ13Cfatty acid, δ13Camino acid, δ15Namino acid, and δ2Hamino acid. Analysis of variance (ANOVA) and linear discriminant analysis (LDA) were used for statistical analysis to compare samples from the USA and China. Parameters that yielded significant variations are δ2HC18:1n-9, δ13CC18:2n-6, δ13CC18:3n-3, δ13CGly, δ13CLeu, δ13CVal, δ2HGlu, δ2HIle, δ2HLeu, and δ2HThr. Our findings suggested that CSIA of fatty acids and amino acids can be useful to differentiate the geographical provenance of walnuts.

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.

A Coding Basis and Three-in-One Integrated Data Visualization Method 'Ana' for the Rapid Analysis of Multidimensional Omics Dataset.

With innovations and advancements in analytical instruments and computer technology, omics studies based on statistical analysis, such as phytochemical omics, oilomics/lipidomics, proteomics, metabolomics, and glycomics, are increasingly popular in the areas of food chemistry and nutrition science. However, a remaining hurdle is the labor-intensive data process because learning coding skills and software operations are usually time-consuming for researchers without coding backgrounds. A MATLAB® coding basis and three-in-one integrated method, 'Ana', was created for data visualizations and statistical analysis in this work. The program loaded and analyzed an omics dataset from an Excel® file with 7 samples * 22 compounds as an example, and output six figures for three types of data visualization, including a 3D heatmap, heatmap hierarchical clustering analysis, and principal component analysis (PCA), in 18 s on a personal computer (PC) with a Windows 10 system and in 20 s on a Mac with a MacOS Monterey system. The code is rapid and efficient to print out high-quality figures up to 150 or 300 dpi. The output figures provide enough contrast to differentiate the omics dataset by both color code and bar size adjustments per their higher or lower values, allowing the figures to be qualified for publication and presentation purposes. It provides a rapid analysis method that would liberate researchers from labor-intensive and time-consuming manual or coding basis data analysis. A coding example with proper code annotations and completed user guidance is provided for undergraduate and postgraduate students to learn coding basis statistical data analysis and to help them utilize such techniques for their future research.

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.

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.

Chardonnay Marc as a New Model for Upcycled Co-products in the Food Industry: Concentration of Diverse Natural Products Chemistry for Consumer Health and Sensory Benefits.

Research continues to provide compelling insights into potential health benefits associated with diets rich in plant-based natural products (PBNPs). Coupled with evidence from dietary intervention trials, dietary recommendations increasingly include higher intakes of PBNPs. In addition to health benefits, PBNPs can drive flavor and sensory perceptions in foods and beverages. Chardonnay marc (pomace) is a byproduct of winemaking obtained after fruit pressing that has not undergone fermentation. Recent research has revealed that PBNP diversity within Chardonnay marc has potential relevance to human health and desirable sensory attributes in food and beverage products. This review explores the potential of Chardonnay marc as a valuable new PBNP ingredient in the food system by combining health, sensory, and environmental sustainability benefits that serves as a model for development of future ingredients within a sustainable circular bioeconomy. This includes a discussion on the potential role of computational methods, including artificial intelligence (AI), in accelerating research and development required to discover and commercialize this new source of PBNPs.

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.