Correction: Ghani et al. Multi-Dimensional Antioxidant Screening of Selected Australian Native Plants and Putative Annotation of Active Compounds. Molecules 2023, 28, 3106.

In the original publication [...].

Multi-Dimensional Antioxidant Screening of Selected Australian Native Plants and Putative Annotation of Active Compounds.

Acacia implexa, Eucalyptus rossii and Exocarpos cupressiformis are native plants of Australia, which were used by the First Peoples for medicinal purposes. In this study, 70% aqueous ethanol crude extracts were prepared from A. implexa bark and leaves, E. rossii leaves and E. cupressiformis leaves, and partitioned via sequential extraction with n-hexane, dichloromethane (DCM), ethyl acetate and ethanol. The crude extracts and fractions were screened for antioxidant activity using a novel, high-throughput lipid-based antioxidant assay, as well as the aqueous ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assay and the Folin-Ciocalteu test for total phenols. In the lipid-based assay, non-polar n-hexane and DCM fractions showed higher antioxidant activity against the formation of peroxides and thiobarbituric acid reactive substances (TBARS) than the other fractions, whereas the non-polar fractions were not effective in aqueous assays. This illustrates that the high potential of the lipid-soluble n-hexane and DCM fractions as antioxidants would have been missed if only aqueous-based assays were used. In addition, the potent antioxidant compounds were putatively annotated using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-qTOF-MS). Gallic acid, (+)-catechin, (-)-epicatechin and tannins were found in most crude extracts.

Optimization and Comparison of Microwave-Assisted Extraction, Supercritical Fluid Extraction, and Eucalyptus Oil-Assisted Extraction of Polycyclic Aromatic Hydrocarbons from Soil and Sediment.

Polycyclic aromatic hydrocarbons (PAHs) are persistent organic compounds of major concern that mainly accumulate in soils and sediments, and their extraction from environmental matrices remains a crucial step when determining the extent of contamination in soils and sediments. The objective of the present study was to compare the extraction of PAHs (phenanthrene, pyrene, chrysene, and benzo[a]pyrene) from spiked soil and sediment using supercritical fluid extraction (SFE) with ethanol as the modifier, microwave-assisted extraction (MAE), and eucalyptus oil-assisted extraction (EuAE). Recoveries of PAHs were comparable between the three methods, and >80% of applied pyrene, chrysene and benzo[a]pyrene were recovered. The most efficient method of extracting PAHs from naturally incurred soils with different levels of contamination was SFE. A longer extraction time was required for the EuAE method compared with SFE and MAE under optimized conditions. However, EuAE required lower extraction temperatures (15-20 °C) compared with SFE (80 °C) and MAE (110-120 °C), and consumed less solvent than SFE and MAE. Compared with hexane/acetone used in MAE, the use of ethanol in SFE and eucalyptus oil in EuAE can be considered as more sustainable approaches to efficiently extract PAHs from spiked/naturally contaminated soils and sediments. And, although less efficient for matrices containing higher carbon content, EuAE offered a cheap, low-tech approach to extracting PAHs. Environ Toxicol Chem 2023;42:982-994. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Eucalyptus saponin- and sophorolipid-mediated desorption of polycyclic aromatic hydrocarbons from contaminated soil and sediment.

The potential for biosurfactant-mediated desorption of polyaromatic hydrocarbons (PAHs) was evaluated using PAH-spiked soil and sediment. PAH desorption behaviors and toxicity of novel saponin biosurfactant extracted from Eucalyptus camaldulensis leaves and sophoro-lipid biosurfactant were investigated. Their PAH desorption efficiencies were compared with rhamnolipid biosurfactant and the industrial-chemical surfactant, Tween 20. Based on the emulsification indices, the salt tolerance of surfactants up to 30 g/L NaCl followed the order of saponin > Tween 20 > sophorolipid > rhamnolipid, while the thermal stability over the range of 15 to 50 °C was in the order of sophorolipid > rhamnolipid > saponin > Tween 20. The saponin biosurfactant emulsion demonstrated the highest stability under a wide range of acidic to basic pHs. PAH extraction percentages of saponin and sophorolipid under the optimized surfactant concentration, volume, and incubation time were 30-50% and 30-70%, respectively. PAH desorption capacities of saponin and sophorolipid were comparable to that of rhamnolipid and Tween 20 for all matrices. Sophorolipid more efficiently desorbed low molecular weight PAHs in soil and sediment compared to the other three surfactants. Microbial respiration was used to determine biosurfactant toxicity to the soil/sediment microbiome and indicated no inhibition of respiration during 60 days of incubation, suggesting that sophorolipid- and saponin-mediated remediation may be sustainable approaches to remove PAHs from contaminated soils and sediments.

Suppression of reductive characters in white wine by Cu fractions: Efficiency and duration of protection during bottle aging.

Cu in wine can suppress sulfidic-odours, but the active forms and duration of protection are uncertain. Additions of 0, 0.3 or 0.6 mg/L Cu(II) were made to Chardonnay and Pinot Grigio at bottling. Throughout a 12- or 14-month storage period, Cu fractions were determined by colorimetry, and sulfhydryl compounds by gas chromatography with sulfur chemiluminescence detection. After Cu(II) addition, the dominant Cu fractions were associated with Cu(II)-organic acids (fraction I) and Cu(I)-thiol complexes (fraction II), and over 8-months their concentrations gradually fell below 0.015 mg/L. During this time, a fraction of Cu, predominantly attributed to sulfide-bound Cu, increased in concentration. Suppression of free hydrogen sulfide was assured when the combined Cu fractions I and II concentrations were above 0.015 mg/L, while free methanethiol suppression required Cu fraction I concentration above 0.035 mg/L. Decay rates for Cu fractions demonstrated that the duration that Cu can actively suppress sulfidic odours is wine-dependent.

Polycyclic aromatic hydrocarbon contamination in soils and sediments: Sustainable approaches for extraction and remediation.

Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic environmental pollutants that are extremely hydrophobic in nature and resistant to biological degradation. Extraction of PAHs from environmental matrices is the first and most crucial step in PAH quantification. Extraction followed by quantification is essential to understand the extent of contamination prior to the application of remediation approaches. Due to their non-polar structures, PAHs can be adsorbed tightly to the organic matter in soils and sediments, making them more difficult to be extracted. Extraction of PAHs can be achieved by a variety of methods. Techniques such as supercritical and subcritical fluid extraction, microwave-assisted solvent extraction, plant oil-assisted extraction and some microextraction techniques provide faster PAH extraction using less organic solvents, while providing a more environmentally friendly and safer process with minimum matrix interferences. More recently, more environmentally friendly methods for soil and sediment remediation have been explored. This often involves using natural chemicals, such as biosurfactants, to solubilize PAHs in contaminated soils and sediments to allow subsequent microbial degradation. Vermiremediation and microbial enzyme-mediated remediation are emerging approaches, which require further development. The following summarises the existing literature on traditional PAH extraction and bioremediation methods and contrasts them to newer, more environmentally friendly ways.

Potential Role of Phenolic Extracts of Mentha in Managing Oxidative Stress and Alzheimer's Disease.

With an increase in the longevity and thus the proportion of the elderly, especially in developed nations, there is a rise in pathological conditions that accompany ageing, such as neurodegenerative disorders. Alzheimer's disease (AD) is a neurodegenerative disease characterized by progressive cognitive and memory decline. The pathophysiology of the disease is poorly understood, with several factors contributing to its development, such as oxidative stress, neuroinflammation, cholinergic neuronal apoptotic death, and the accumulation of abnormal proteins in the brain. Current medications are only palliative and cannot stop or reverse the progression of the disease. Recent clinical trials of synthetic compounds for the treatment of AD have failed because of their adverse effects or lack of efficacy. Thus, there is impetus behind the search for drugs from natural origins, in addition to the discovery of novel, conventional therapeutics. Mints have been used traditionally for conditions relevant to the central nervous system. Recent studies showed that mint extracts and/or their phenolic constituents have a neuroprotective potential and can target multiple events of AD. In this review, we provide evidence of the potential role of mint extracts and their derivatives as possible sources of treatments in managing AD. Some of the molecular pathways implicated in the development of AD are reviewed, with focus on apoptosis and some redox pathways, pointing to mechanisms that may be modulated for the treatment of AD, and the need for future research invoking knowledge of these pathways is highlighted.

Neuroprotective Activity of Mentha Species on Hydrogen Peroxide-Induced Apoptosis in SH-SY5Y Cells.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with an unclear cause. It appears that multiple factors participate in the process of neuronal damage including oxidative stress and accumulation of the protein amyloid ß (Aß) in the brain. The search for a treatment for this disorder is essential as current medications are limited to alleviating symptoms and palliative effects. The aim of this study is to investigate the effects of mint extracts on selected mechanisms implicated in the development of AD. To enable a thorough investigation of mechanisms, including effects on ß-secretase (the enzyme that leads to the formation of Aß), on Aß aggregation, and on oxidative stress and apoptosis pathways, a neuronal cell model, SH-SY5Y cells, was selected. Six Mentha taxa were investigated for their in vitro ß-secretase (BACE) and Aß-aggregation inhibition activities. Moreover, their neuroprotective effects on H2O2-induced oxidative stress and apoptosis in SH-SY5Y cells were evaluated through caspase activity. Real-time PCR and Western blot analysis were carried out for the two most promising extracts to determine their effects on signalling pathways in SH-SY5Y cells. All mint extracts had strong BACE inhibition activity. M. requienii extracts showed excellent inhibition of Aß-aggregation, while other extracts showed moderate inhibition. M. diemenica and M. requienii extracts lowered caspase activity. Exposure of SH-SY5Y cells to M. diemenica extracts resulted in a decrease in the expression of pro-apoptotic protein, Bax, and an elevation in the anti-apoptotic protein, Bcl-xL, potentially mediated by down-regulation of the ASK1-JNK pathway. These results indicate that mint extracts could prevent the formation of Aß and also could prevent their aggregation if they had already formed. M. diemenica and M. requienii extracts have potential to suppress apoptosis at the cellular level. Hence, mint extracts could provide a source of efficacious compounds for a therapeutic approach for AD.

Bulk and compound-specific stable isotope ratio analysis for authenticity testing of organically grown tomatoes.

Until now, there has been a lack of analytical methods that can reliably verify the authenticity of organically grown plants and derived organic food products. In this study, stable isotope ratio analysis of hydrogen (H, δ2H), carbon (C, δ13C), nitrogen (N, δ15N), oxygen (O, δ18O) and sulfur (S, δ34S) was conducted along the tomato passata production process using organic and conventionally grown tomatoes from two Italian regions over two years. A gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) based method was developed and applied for analysis of C and N isotope ratios in amino acids derived from tomatoes. Of the bulk isotope ratios, δ15N was the most significant parameter for discriminating organic from conventional products. The classification power was improved significantly by compound-specific isotope analysis regardless of the production years and regions. We conclude that isotope analysis of amino acids is a novel analytical tool for complementing existing certification and control procedures in the organic tomato sector.

Different Processing Practices and the Frying Life of Refined Canola Oil.

Refined expeller-pressed (RCanO-I and RCanO-II) and expeller-pressed and solvent-extracted blended (RCanO-III and RCanO-IV) canola oils were compared to determine the effect of processing (extraction) practice on the frying life of canola oil. Samples were from the 2016/2017 and 2017/2018 production seasons and were used to fry potato chips for 36 to 48 cycles. Frying life was assessed by the total polar compounds, retention of tocopherols, antioxidant activity, and other quality indices. RCanO-II exhibited significantly, the longest frying life as compared with the other three oils and this correlated with tocopherol retention and antioxidant activity (p < 0.05). The extraction practice influenced the frying life of canola oil, but this was dependent on other processing practices employed by the individual processors. Variations in initial oil quality dictated the rates of chemical reactions occurring in the oils during frying and influenced oil stability.

Development of a Method Suitable for High-Throughput Screening to Measure Antioxidant Activity in a Linoleic Acid Emulsion.

An improved system for measuring antioxidant activity via thiobarbituric acid reactive substances and ferric thiocyanate assays is reported, on the basis of oxidation of a linoleic acid (LA) emulsion. Oxidation times were reduced from 20 h to 5 h by increasing the reaction temperature from 37 °C to 50 °C and with an acceptable precision of <10% coefficient of variation (CV). Antioxidants varying in polarity and chemical class-250 µM Trolox, quercetin, ascorbic acid and gallic acid-were used for method optimisation. Further reductions in reaction time were investigated through the addition of catalysts, oxygen initiators or increasing temperature to 60 °C; however, antioxidant activity varied from that established at 37 °C and 20 h reaction time-the method validation conditions. Further validation of the method was achieved with catechin, epicatechin, caffeic acid and α-tocopherol, with results at 50 °C and 5 h comparable to those at 37 °C and 20 h. The improved assay has the potential to rapidly screen antioxidants of various polarities, thus making it useful in studies where large numbers of plant extracts require testing. Furthermore, as this assay involves protection of a lipid, the assay is likely to provide complementary information to well-established tests, such as the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay.

Differentiation of wood-derived vanillin from synthetic vanillin in distillates using gas chromatography/combustion/isotope ratio mass spectrometry for δ13 C analysis.

RATIONALE: Typical storage in oak barrels releases in distillates different degradation products such as vanillin, which play an important role in flavour and aroma. The addition of vanillin, as well as other aroma compounds, of different origin is prohibited by European laws. As vanillin samples from different sources have different δ13 C values, the δ13 C value could be used to determine whether the vanillin is authentic (lignin-derived), or if it has been added from another source (e.g. synthetic). METHODS: The δ13 C values for vanillin derived from different sources, including natural, synthetic and tannins, were measured by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS), after diethyl ether addition and/or ethanol dilution. A method for analysing vanillin in distillates after dichloromethane extraction was developed. Tests were undertaken to prove the reliability, reproducibility and accuracy of the method with standards and samples. Distillate samples were run to measure the δ13 C values of vanillin and to compare them with values for other sources of vanillin. RESULTS: δ13 C values were determined for: natural vanillin extracts (-21.0 to -19.3‰, 16 samples); vanillin ex-lignin (-28.2‰, 1 sample); and synthetic vanillin (-32.6 to -29.3‰, 7 samples). Seventeen tannin samples were found to have δ13 C values of -29.5 to -26.7‰, which were significantly different (p < 0.05) from those of the natural and synthetic vanillins. The vanillin δ13 C values measured in distillates (-28.9 to -25.7‰) were mainly in the tannin range, although one spirit (-32.5‰) was found to contain synthetic vanillin. CONCLUSIONS: The results show that synthetic vanillin added to a distillate could be differentiated from vanillin derived from oak barrels by their respective δ13 C values. The GC/C/IRMS method could be a useful tool in the determination of adulteration of distillates.

Dietary Effects on Stable Carbon Isotope Composition of Fatty Acids in Polar and Neutral Fractions of Intramuscular Fat of Lambs.

In this study we measured δ13C values of the main fatty acids (FA) present in neutral and polar intramuscular lipids of meat samples from 24 lambs, fed with four different diets supplemented with sunflower and linseed oil and the tanniferous shrub Cistus ladanifer L. The objective was to understand if the increase in intramuscular fat observed in lambs fed simultaneously C. ladanifer and oil was explained mostly by incorporation of diet derived FA or by increased de novo FA synthesis. De novo FA synthesis was evaluated by 13C enrichment (‰) of 16:0 in intramuscular lipids compared to bulk diet or compared to dietary 16:0. Oil reduced 13C enrichment of 16:0 in muscle lipid but had no effect when the diet included C. ladanifer (P value <0.01). Thus, dietary C. ladanifer blocked the inhibitory effects of lipid supplementation on de novo FA synthesis.

Measurement of antioxidant activity with the thiobarbituric acid reactive substances assay.

The thiobarbituric acid reactive substances (TBARS) assay is widely used to measure lipid oxidation and antioxidant activity in food and physiological systems. However, there has been no review (to our knowledge) that focuses exclusively on this test. This review presents an overview of the current use of the TBARS test in food and physiological systems, before looking at the various ways in which the assay is used in studies on antioxidant activity. As an antioxidant assay, the TBARS test may lack acceptable reproducibility, and long reaction times may preclude its adoption as a rapid screening method. Despite these potential limitations, there are features of the TBARS test that make it useful as a complement to popular screening tests such as Trolox equivalent antioxidant capacity. This review concludes with proposals for development of the TBARS test so that it can be used as a rapid and robust antioxidant assay.

A rapid method for the simultaneous quantification of the major tocopherols, carotenoids, free and esterified sterols in canola (Brassica napus) oil using normal phase liquid chromatography.

A normal phase high performance liquid chromatography (HPLC) method was developed to simultaneously quantify several prominent bioactive compounds in canola oil vis. α-tocopherol, γ-tocopherol, δ-tocopherol, ß-carotene, lutein, ß-sitosterol, campesterol and brassicasterol. The use of sequential diode array detection (DAD) and tandem mass spectrometry (MS/MS) allowed direct injection of oils, diluted in hexane without derivatisation or saponification, greatly reducing sample preparation time, and permitting the quantification of both free sterols and intact sterol esters. Further advantages over existing methods included increased analytical selectivity, and a chromatographic run time substantially less than other reported normal phase methods. The HPLC-DAD-MS/MS method was applied to freshly extracted canola oil samples as well as commercially available canola, palm fruit, sunflower and olive oils.

The decay of ascorbic acid in a model wine system at low oxygen concentration.

The present study investigated the impact of temperature on the degradation of ascorbic acid in low oxygen conditions in a model white wine. The concentrations of ascorbic acid, furfural, sulfur dioxide and phenolic-type products were monitored in a model white wine stored under non-oxidative conditions at 45.0, 36.5 and 24.0 °C for up to 693 days. The concentrations of both ascorbic acid and sulfur dioxide decreased over the analysis period while furfural and other colourless phenolic products increased in concentration, despite the presence of residual sulfur dioxide. The decay of ascorbic acid in the low oxygen conditions followed first-order kinetics and the rate constants were determined to be (3.5±0.2)×10(-8), (1.02±0.07)×10(-8), and (0.184±0.009)×10(-8) s(-1) for 45.0, 36.5 and 24.0 °C (n=5, standard error), respectively, and the activation energy was 110±3 kJ/mol (n=3, standard error). Importantly, these data allow more accurate prediction of the temperature-induced loss of ascorbic acid in low oxygen conditions during transport or storage of wine.

Total phenolic content, antioxidant activity, and cross-cultural consumer rejection threshold in white and red wines functionally enhanced with catechin-rich extracts.

White and red wines spiked with catechin-rich green tea extract and grape seed extract were assessed for phenolic content, antioxidant activity, and cross-cultural consumer rejection thresholds in relation to wine as a functional food. Health functionality is an important factor in functional foods, and spiking pure compounds or plant extracts is an effective method to increase or control functionality. The total phenolic content and antioxidant activity were measured in wines spiked to different extract concentrations, namely, control and 50, 100, 200, 400, and 800 mg/L, to confirm the dose-response curves in both white and red wines. Consumer rejection thresholds (CRTs) were established for spiked wines in a Korean and in an Australian population. Our results showed that the green tea extract and grape seed extract increased the antioxidant activity dose dependently, and the CRTs varied considerably between the Korean and the Australian groups, with Koreans preferring wines spiked with green tea extract and Australians showing a preference for wines spiked with grape seed extract. These results have implications for producing wine products that are enhanced in phenolic compounds and targeted to different cultural groups.

Bioprospecting traditional Pakistani medicinal plants for potent antioxidants.

Antioxidant potential of four methanol extracts from three selected plant species, namely Salvia nubicola (Lamiaceae), Acer oblongifolium (Aceraceae) and Hedera nepalensis (Araliaceae) was measured using assays in aqueous and lipid systems. Antioxidant activities were investigated in aqueous systems by using DPPH radical-scavenging assay, ABTS radical-scavenging assay and DNA protection assay, while antioxidant activity in a lipid system was determined by using the thiobarbituric acid-reactive substances (TBARS) assay. Additionally, the Folin-Ciocalteu method was used to measure total phenolic content. Methanol extracts of leaves and flowers of S. nubicola showed the highest Trolox equivalent (TE) values in the case of the DPPH assay, 2484±4.9mmol TE/g extract, as well as total phenolic content, 139±0.2mg gallic acid equivalents/g extract. Three fractions (A-C) of the methanol extract of S. nubicola leaves and flowers were produced by semi-preparative HPLC. Fraction B was found to be the most active in the DPPH radical-scavenging assay and had the highest total phenol content. HPLC-DAD and LC-MS revealed rosmarinic acid in S. nubicola extracts and chlorogenic acid and rutin in H. nepalensis extracts as the main phenolic antioxidants.

Ascorbic acid: a review of its chemistry and reactivity in relation to a wine environment.

Extensive reviews of research are available on the use of ascorbic acid, and its consequent degradation pathways, in physiological conditions or food matrices. However, very little information can be found for wine-related systems. This review highlights the relevant chemistry and reactivity of ascorbic acid with a focus on its behavior and potential behavior in a wine environment. The review describes the use of ascorbic acid as a complementary antioxidant preservative to sulfur dioxide along with the metal-catalyzed and radical-dependent manner by which it achieves this role. The relevant degradation products of ascorbic acid in aerobic and anaerobic conditions are presented as well as the interaction of these degradation products with sulfur dioxide and other wine-relevant sulfur compounds. Limitations in existing knowledge, especially regarding the crossover between the antioxidant and pro-oxidant roles of ascorbic acid, are identified.

Antioxidant action of glutathione and the ascorbic acid/glutathione pair in a model white wine.

Glutathione was assessed individually, and in combination with ascorbic acid, for its ability to act as an antioxidant with respect to color development in an oxidizing model white wine system. Glutathione was utilized at concentrations normally found in wine (30 mg/L), as well as at concentrations 20-fold higher (860 mg/L), the latter to afford ascorbic acid (500 mg/L) to glutathione ratios of 1:1. The model wine systems were stored at 45 °C without sulfur dioxide and at saturated oxygen levels, thereby in conditions highly conducive to oxidation. Under these conditions the results demonstrated the higher concentration of glutathione could initially provide protection against oxidative coloration, but eventually induced color formation. In the period during which glutathione offered a protective effect, the production of xanthylium cation pigment precursors and o-quinone-derived phenolic compounds was limited. When glutathione induced coloration, polymeric pigments were formed, but these were different from those found in model wine solutions without glutathione. In the presence of ascorbic acid, high concentrations of glutathione were able to delay the decay in ascorbic acid and inhibit the reaction of ascorbic acid degradation products with the wine flavanol compound (+)-catechin. However, on depletion, the glutathione again induced the production of a range of different polymeric pigments. These results highlight new mechanisms through which glutathione can offer both protection and spoilage during the oxidative coloration of a model wine.