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Introduction: Accurately identifying and quantifying polar metabolites using untargeted metabolomics has proven challenging in comparison to mid to non-polar metabolites. Hydrophilic interaction chromatography and gas chromatography–mass spectrometry are predominantly used to target polar metabolites. Objectives: This study aims to demonstrate a simple one-step extraction combined with liquid chromatography–mass spectrometry (LC–MS) that reliably retains polar metabolites. Methods: The method involves a MilliQ + 10% trichloroacetic acid extraction from 6 healthy individuals serum, combined with porous graphitic carbon liquid chromatography–mass spectrometry (LC–MS). The coefficient of variation (CV) assessed retention reliability of polar metabolites with logP as low as − 9. QreSS (Quantification, Retention, and System Suitability) internal standards determined the method's consistency and recovery efficiency. Results: The method demonstrated reliable retention (CV < 0.30) of polar metabolites within a logP range of − 9.1 to 5.6. QreSS internal standards confirmed consistent performance (CV < 0.16) and effective recovery (70–130%) of polar to mid-polar metabolites. Quality control dilution series demonstrated that ~ 80% of annotated metabolites could be accurately quantified (Pearson's correlation coefficient > 0.80) within their concentration range. Repeatability was demonstrated through clustering of repeated extractions from a single sample. Conclusion: This LC–MS method is better suited to covering the polar segment of the metabolome than current methods, offering a reliable and efficient approach for accurate quantification of polar metabolites in untargeted metabolomics. [ABSTRACT FROM AUTHOR]

As obesity develops, metabolic changes increase the risk of non-communicable diseases such as type 2 diabetes (T2D). Weight loss is crucial for improving health in T2D and cardiometabolic conditions. However, weight loss rates vary between individuals, even with identical diets or energy restrictions, highlighting the need to identify markers or predictors of weight loss success to enhance intervention outcomes. Using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics, we investigated the change in serum polar metabolites in 28 women with overweight or obesity and prediabetes who completed an 8-week low-energy diet (LED) as part of the PREVIEW (PREVention of diabetes through lifestyle intervention and population studies in Europe and around the World) clinical trial. We aimed to characterize the metabolic shift in substrate oxidation under fixed energy intake (~4 MJ/day) and its relation to weight loss success. Nine of the thirty-four serum metabolites identified significantly changed during the LED phase: 3-hydroxybutyrate, O-acetylcarnitine, 2-hydroxybutyrate, mannose, dimethyl sulfone and isobutyrate increased, whilst choline, creatine and tyrosine decreased. These results confirmed a shift towards lipid oxidation, but no metabolites predicted the response to the LED-induced weight loss. Further studies in larger populations are required to validate these metabolites as biomarkers of diet exposure. [ABSTRACT FROM AUTHOR]

Depression will be the disease with the highest incidence worldwide by 2030. Data indicate that postmenopausal women have a higher incidence of mood disorders, and this high vulnerability seems to be related to hormonal changes and weight gain. Although research evaluating the profile of metabolites in mood disorders is advancing, further research, maintaining consistent methodology, is necessary to reach a consensus. Therefore, the objective of the present study was to carry out an exploratory analysis of the plasma polar metabolites of pre- and postmenopausal women to explore whether the profile is affected by depression. The plasma analysis of 50 polar metabolites was carried out in a total of 67 postmenopausal women, aged between 50 and 65 years, either without depression (n = 25) or with depression symptoms (n = 42), which had spontaneous onset of menopause and were not in use of hormone replacement therapy, insulin, or antidepressants; and in 42 healthy premenopausal women (21 without depression and 21 with depression symptoms), aged between 40 and 50 years and who were not in use of contraceptives, insulin, or antidepressants. Ten metabolites were significantly affected by depression symptoms postmenopause, including adenosine (FDR = 3.778 × 10−14), guanosine (FDR = 3.001 × 10−14), proline (FDR = 1.430 × 10−6), citrulline (FDR = 0.0001), lysine (FDR = 0.0004), and carnitine (FDR = 0.0331), which were down-regulated, and dimethylglycine (FDR = 0.0022), glutathione (FDR = 0.0048), creatine (FDR = 0.0286), and methionine (FDR = 0.0484) that were up-regulated. In premenopausal women with depression, oxidized glutathione (FDR = 0.0137) was down-regulated, and dimethylglycine (FDR = 0.0406) and 4-hydroxyproline (FDR = 0.0433) were up-regulated. The present study provided new data concerning the consequences of depression on plasma polar metabolites before and after the establishment of menopause. The results demonstrated that the postmenopausal condition presented more alterations than the premenopausal period and may indicate future measures to treat the disturbances involved in both menopause and depression. [ABSTRACT FROM AUTHOR]

Lung cancer is a common malignancy that is frequently associated with systemic metabolic disorders. Early detection is pivotal to survival improvement. Although blood biomarkers have been used in its early diagnosis, missed diagnosis and misdiagnosis still exist due to the heterogeneity of lung cancer. Integration of multiple biomarkers or trans-omics results can improve the accuracy and reliability for lung cancer diagnosis. As metabolic reprogramming is a hallmark of lung cancer, metabolites, specifically lipids might be useful for lung cancer detection, yet systematic characterizations of metabolites in lung cancer are still incipient. The present study profiled the polar metabolome and lipidome in the plasma of lung cancer patients to construct an inclusive metabolomic atlas of lung cancer. A comprehensive analysis of lung cancer was also conducted combining metabolomics with clinical phenotypes. Furthermore, the differences in plasma lipid metabolites were compared and analyzed among different lung cancer subtypes. Alcohols, amides, and peptide metabolites were significantly increased in lung cancer, while carboxylic acids, hydrocarbons, and fatty acids were remarkably decreased. Lipid profiling revealed a significant increase in plasma levels of CER, PE, SM, and TAG in individuals with lung cancer as compared to those in healthy controls. Correlation analysis confirmed the association between a panel of metabolites and TAGs. Clinical trans-omics studies elucidated the complex correlations between lipidomic data and clinical phenotypes. The present study emphasized the clinical importance of lipidomics in lung cancer, which involves the correlation between metabolites and the expressions of other omics, ultimately influencing clinical phenotypes. This novel trans-omics network approach would facilitate the development of precision therapy for lung cancer. 1. Integrating multiple biomarkers or trans-omics results improves diagnostic accuracy and reliability in heterogeneous lung cancer. 2. Metabolomics and lipidomics, along with clinical phenotypes, construct a comprehensive metabolic profile of lung cancer patients. 3. TAG expression shows strong positive correlation with polar metabolites, potentially impacting clinical phenotypic changes in lung cancer patients. [ABSTRACT FROM AUTHOR]

As obesity develops, metabolic changes increase the risk of non-communicable diseases such as type 2 diabetes (T2D). Weight loss is crucial for improving health in T2D and cardiometabolic conditions. However, weight loss rates vary between individuals, even with identical diets or energy restrictions, highlighting the need to identify markers or predictors of weight loss success to enhance intervention outcomes. Using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics, we investigated the change in serum polar metabolites in 28 women with overweight or obesity and prediabetes who completed an 8-week low-energy diet (LED) as part of the PREVIEW (PREVention of diabetes through lifestyle intervention and population studies in Europe and around the World) clinical trial. We aimed to characterize the metabolic shift in substrate oxidation under fixed energy intake (~4 MJ/day) and its relation to weight loss success. Nine of the thirty-four serum metabolites identified significantly changed during the LED phase: 3-hydroxybutyrate, O-acetylcarnitine, 2-hydroxybutyrate, mannose, dimethyl sulfone and isobutyrate increased, whilst choline, creatine and tyrosine decreased. These results confirmed a shift towards lipid oxidation, but no metabolites predicted the response to the LED-induced weight loss. Further studies in larger populations are required to validate these metabolites as biomarkers of diet exposure.

An untargeted Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics approach was applied as a first attempt to explore the metabolome of pigs treated with antibiotics. The final goal was to investigate the possibility of discriminating between antibiotic-treated (TX group) and untreated pigs (CTRL group), with the further perspective of identifying the authentication tools for antibiotic-free pork supply chains. In particular, 41 samples of pig liver were subjected to a biphasic extraction to recover both the polar and the non-polar metabolites, and the 1H NMR spectroscopy analysis was performed on the two separate extracts. Unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate statistical analysis of 1H NMR spectra data in the range 0–9 ppm provided metabolomic fingerprinting useful for the discrimination of pig livers based on the antibiotic treatment to which they were exposed. Moreover, within the signature patterns, significant discriminating metabolites were identified among carbohydrates, choline and derivatives, amino acids and some lipid-class molecules. The encouraging findings of this exploratory study showed the feasibility of the untargeted metabolomic approach as a novel strategy in the authentication framework of pork supply chains and open a new horizon for a more in-depth investigation. [ABSTRACT FROM AUTHOR]

Seed-to-seedling transition plays a crucial role in plant vegetation. However, changes in the metabolome of crop seedlings during seed germination and early seedling development are mostly unknown and require a deeper explanation. The present study attempted to compare qualitative and quantitative changes in polar metabolites during the seed germination and early development of seedlings of three different and important crop types: pea, cucumber, and wheat. The application of gas chromatography coupled with a flame ionization detector, as well as gas chromatography coupled with mass spectrometry, identified 51 polar metabolites. During seed imbibition/germination, the rapid degradation of raffinose family oligosaccharides (RFOs) preceded a dramatic increase in the concentrations of intermediates of glycolysis and the TCA cycle in embryonic axes (of pea and cucumber) or embryos (of wheat), confirming the important role of RFOs in the resumption of respiration and seed-to-seedling transition. After germination, the metabolic profiles of the growing roots, epicotyl/hypocotyl/coleoptile, and cotyledons/endosperm changed according to fluctuations in the concentrations of soluble carbohydrates, amino acids, and organic acids along the timeline of seedling growth. Moreover, the early increase in species-specific metabolites justified their role in seedling development owing to their participation in nitrogen metabolism (homoserine in pea), carbon translocation (galactinol, raffinose, and stachyose), and transitory carbon accumulation (1-kestose in wheat). The obtained metabolic profiles may constitute an important basis for further research on seedling reactions to stress conditions, including identification of metabolic markers of stress resistance. [ABSTRACT FROM AUTHOR]

Depression will be the disease with the highest incidence worldwide by 2030. Data indicate that postmenopausal women have a higher incidence of mood disorders, and this high vulnerability seems to be related to hormonal changes and weight gain. Although research evaluating the profile of metabolites in mood disorders is advancing, further research, maintaining consistent methodology, is necessary to reach a consensus. Therefore, the objective of the present study was to carry out an exploratory analysis of the plasma polar metabolites of pre- and postmenopausal women to explore whether the profile is affected by depression. The plasma analysis of 50 polar metabolites was carried out in a total of 67 postmenopausal women, aged between 50 and 65 years, either without depression (n = 25) or with depression symptoms (n = 42), which had spontaneous onset of menopause and were not in use of hormone replacement therapy, insulin, or antidepressants; and in 42 healthy premenopausal women (21 without depression and 21 with depression symptoms), aged between 40 and 50 years and who were not in use of contraceptives, insulin, or antidepressants. Ten metabolites were significantly affected by depression symptoms postmenopause, including adenosine (FDR = 3.778 × 10−14), guanosine (FDR = 3.001 × 10−14), proline (FDR = 1.430 × 10−6), citrulline (FDR = 0.0001), lysine (FDR = 0.0004), and carnitine (FDR = 0.0331), which were down-regulated, and dimethylglycine (FDR = 0.0022), glutathione (FDR = 0.0048), creatine (FDR = 0.0286), and methionine (FDR = 0.0484) that were up-regulated. In premenopausal women with depression, oxidized glutathione (FDR = 0.0137) was down-regulated, and dimethylglycine (FDR = 0.0406) and 4-hydroxyproline (FDR = 0.0433) were up-regulated. The present study provided new data concerning the consequences of depression on plasma polar metabolites before and after the establishment of menopause. The results demonstrated that the postmenopausal condition presented more alterations than the premenopausal period and may indicate future measures to treat the disturbances involved in both menopause and depression.

Mammalian hibernators undergo substantial changes in metabolic function throughout the seasonal hibernation cycle. We report here the polar metabolomic profile of white adipose tissue isolated from active and hibernating thirteen-lined ground squirrels (Ictidomys tridecemlineatus). Polar compounds in white adipose tissue were extracted from five groups representing different timepoints throughout the seasonal activity-torpor cycle and analyzed using hydrophilic interaction liquid chromatography-mass spectrometry in both the positive and negative ion modes. A total of 224 compounds out of 660 features detected after curation were annotated. Unsupervised clustering using principal component analysis revealed discrete clusters representing the different seasonal timepoints throughout hibernation. One-way analysis of variance and feature intensity heatmaps revealed metabolites that varied in abundance between active and torpid timepoints. Pathway analysis compared against the KEGG database demonstrated enrichment of amino acid metabolism, purine metabolism, glycerophospholipid metabolism, and coenzyme A biosynthetic pathways among our identified compounds. Numerous carnitine derivatives and a ketone that serves as an alternate fuel source, beta-hydroxybutyrate (BHB), were among molecules found to be elevated during torpor. Elevated levels of the BHB-carnitine conjugate during torpor suggests the synthesis of beta-hydroxybutyrate in white adipose mitochondria, which may contribute directly to elevated levels of circulating BHB during hibernation. [ABSTRACT FROM AUTHOR]

Understanding the chemical nature of wine aroma demands accurate quantitative determinations of different odor-active compounds. Quantitative determinations of enolones (maltol, furaneol, homofuraneol, and sotolon) and vanillin derivatives (vanillin, methyl vanillate, ethyl vanillate, and acetovanillone) at low concentrations are complicated due to their high polarity. For this reason, this paper presents an improved and automated version for the accurate measure of these common trace wine polar compounds (enolones and vanillin derivatives). As a result, a faster and more user-friendly method with a reduction of organic solvents and resins was developed and validated. The optimization of some stages of the solid phase extraction (SPE) process, such as washing with an aqueous solution containing 1% NaHCO3 at pH 8, led to cleaner extracts and solved interference problems. Due to the polarity of these type of compounds, an optimization of the large volume injection was also carried out. Finally, a programmable temperature vaporization (PTV) quartz glass inlet liner without wool was used. The injector temperature was raised to 300 °C in addition to applying a pressure pulse of 180 kPa for 4 min. Matrix effects were solved by the use of adequate internal standards, such as ethyl maltol and 3′,4′-(methylenedioxy)acetophenone. Method figures of merit were highly satisfactory: good linearity (r2 > 0.98), precision (relative standard deviation, RSD < 10%), high recovery (RSD > 89%), and low detection limits (<0.7 μg/L). Enolones and vanillin derivatives are associated with wine aging. For this reason, the methodology was successfully applied to the quantification of these compounds in 16 Spanish red wines and 12 mistelles. Odor activity values (OAV) indicate that furaneol should be considered an aroma impact odorant in red wines and mistelles (OAV > 1) while homofuraneol and sotolon could also produce changes in their aroma perceptions (0.1 < OAV < 1). [ABSTRACT FROM AUTHOR]

Harvested avocado fruits can be potentially exposed both to pre- and post-harvest factors that may influence the susceptibility of fruit to quality loss. Regular air (RA) storage consists of approximately 78% nitrogen, 21% oxygen, 0.3% carbon dioxide, and smaller amounts of some other gasses. A controlled atmosphere (CA) is a changed atmosphere that is used to delay respiration and reduce fungal and physiological deterioration, and it is an appropriate and practical technology for prolonging the shelf life and maintaining the quality of fruits on a large scale. An experiment was conducted by collecting fruits from two different avocado orchards (Bartolillo and Quilhuica) during two harvest seasons based on dry matter content and two subsequent years. The current study was designed to investigate the changes in polar metabolites, phenolic compounds, and fatty acids in the skin of Hass avocados stored under two distinct conditions. Bartolillo orchard fruits were mostly correlated to linoleic and oleic acid as important variables. For Quilhuica, fruits were mostly correlated with palmitoleic, palmitic, and oleic acids. Phenolic content increased at the beginning of storage and decreased at the end of storage for one orchard and contrarily for others, indicating that the result was dependent on the orchard and storage condition. Serine, glutaric acid, xylitol and D-mannitol were the polar metabolites that most correlated with fruits of the Quilhuica orchard while ß-sitosterol and gluconic were related to fruits of the Bartolillo orchard. The differences in fatty acids, polar metabolites and phenolics were dependent on orchard and storage conditions. [ABSTRACT FROM AUTHOR]

The choice of rootstock has a significant impact, not only on fruit growth and development, but also on avocado fruit quality and postharvest performance. The objective of this study was to evaluate and compare attributes related to the postharvest quality of Hass avocados from "Mexicola" and "Duke 7" rootstocks grown under similar conditions. This study included two harvests: early (23–26% dry matter) and middle (>26–30% dry matter) per season for the 2016/2017 and 2017/2018 seasons and two storage conditions (regular air (RA) at 5 °C and controlled atmosphere (CA) at 4 kPa O2 and 6 kPa CO2 at 5 °C) for 55 days. The results showed significant differences in firmness, color and vascular and flesh browning between storage conditions; in addition to these last three attributes, the rootstock played an important role. The fatty acid profile did not reveal significant differences between fruit from both rootstocks. Finally, the polar metabolite profiles revealed differences only for the storage condition, not associated to the rootstock, which could affect the postharvest performance of Hass avocado fruit. This study is one of the few available showing the interaction of rootstock/Hass cultivar on fruit quality and postharvest performance. [ABSTRACT FROM AUTHOR]

Mercury (Hg) is a dangerous and persistent trace element. Its organic and highly toxic form, methylmercury (MeHg), easily crosses biological membranes and accumulates in biota. Nevertheless, understanding the mechanisms of dietary MeHg toxicity in fish remains a challenge. A time-course experiment was conducted with juvenile white seabreams, Diplodus sargus (Linnaeus, 1758), exposed to realistic levels of MeHg in feed (8.7 μg g−1, dry weight), comprising exposure (E; 7 and 14 days) and post-exposure (PE; 28 days) periods. Total Hg levels increased with time in gills and liver during E and decreased significantly in PE (though levels of control fish were reached only for gills), with liver exhibiting higher levels (2.7 times) than gills. Nuclear magnetic resonance (NMR)-based metabolomics revealed multiple and often differential metabolic changes between fish organs. Gills exhibited protein catabolism, disturbances in cholinergic neurotransmission, and changes in osmoregulation and lipid and energy metabolism. However, dietary MeHg exposure provoked altered protein metabolism in the liver with decreased amino acids, likely for activation of defensive strategies. PE allowed for the partial recovery of both organs, even if with occurrence of oxidative stress and changes of energy metabolism. Overall, these findings support organ-specific responses according to their sensitivity to Hg exposure, pointing out that indications obtained in biomonitoring studies may depend also on the selected organ. [ABSTRACT FROM AUTHOR]

Accumulation of anthocyanins in detached leaves and in excised stems of Kalanchoë blossfeldiana kept under natural light conditions in the presence or absence of methyl jasmonate (JA-Me) was investigated. When the abaxial surface of detached leaves was held lower than the adaxial surface (the normal or natural position) under natural light conditions, anthocyanins were not accumulated on the abaxial side of the leaves. In contrast, when the adaxial surface of detached leaves was held lower than the abaxial surface (inverted position), anthocyanins were highly accumulated on the abaxial side of the leaves. These phenomena were independent of the growth stage of K. blossfeldiana as well as photoperiod. Application of JA-Me in lanolin paste significantly inhibited anthocyanin accumulation induced on the abaxial side of detached leaves held in an inverted position in a dose-dependent manner. Anthocyanin accumulation in the excised stem in response to natural light was also significantly inhibited by JA-Me in lanolin paste. Possible mechanisms of anthocyanin accumulation on the abaxial side of detached K. blossfeldiana leaves held in an inverted position under natural light conditions and the inhibitory effect of JA-Me on this process are described. The accompanying changes in the content of primary metabolites and histological analyses were also described. [ABSTRACT FROM AUTHOR]

Mammalian hibernators undergo substantial changes in metabolic function throughout the seasonal hibernation cycle. We report here the polar metabolomic profile of white adipose tissue isolated from active and hibernating thirteen-lined ground squirrels (Ictidomys tridecemlineatus). Polar compounds in white adipose tissue were extracted from five groups representing different timepoints throughout the seasonal activity-torpor cycle and analyzed using hydrophilic interaction liquid chromatography-mass spectrometry in both the positive and negative ion modes. A total of 224 compounds out of 660 features detected after curation were annotated. Unsupervised clustering using principal component analysis revealed discrete clusters representing the different seasonal timepoints throughout hibernation. One-way analysis of variance and feature intensity heatmaps revealed metabolites that varied in abundance between active and torpid timepoints. Pathway analysis compared against the KEGG database demonstrated enrichment of amino acid metabolism, purine metabolism, glycerophospholipid metabolism, and coenzyme A biosynthetic pathways among our identified compounds. Numerous carnitine derivatives and a ketone that serves as an alternate fuel source, beta-hydroxybutyrate (BHB), were among molecules found to be elevated during torpor. Elevated levels of the BHB-carnitine conjugate during torpor suggests the synthesis of beta-hydroxybutyrate in white adipose mitochondria, which may contribute directly to elevated levels of circulating BHB during hibernation.

Seed-to-seedling transition plays a crucial role in plant vegetation. However, changes in the metabolome of crop seedlings during seed germination and early seedling development are mostly unknown and require a deeper explanation. The present study attempted to compare qualitative and quantitative changes in polar metabolites during the seed germination and early development of seedlings of three different and important crop types: pea, cucumber, and wheat. The application of gas chromatography coupled with a flame ionization detector, as well as gas chromatography coupled with mass spectrometry, identified 51 polar metabolites. During seed imbibition/germination, the rapid degradation of raffinose family oligosaccharides (RFOs) preceded a dramatic increase in the concentrations of intermediates of glycolysis and the TCA cycle in embryonic axes (of pea and cucumber) or embryos (of wheat), confirming the important role of RFOs in the resumption of respiration and seed-to-seedling transition. After germination, the metabolic profiles of the growing roots, epicotyl/hypocotyl/coleoptile, and cotyledons/endosperm changed according to fluctuations in the concentrations of soluble carbohydrates, amino acids, and organic acids along the timeline of seedling growth. Moreover, the early increase in species-specific metabolites justified their role in seedling development owing to their participation in nitrogen metabolism (homoserine in pea), carbon translocation (galactinol, raffinose, and stachyose), and transitory carbon accumulation (1-kestose in wheat). The obtained metabolic profiles may constitute an important basis for further research on seedling reactions to stress conditions, including identification of metabolic markers of stress resistance.

An untargeted Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics approach was applied as a first attempt to explore the metabolome of pigs treated with antibiotics. The final goal was to investigate the possibility of discriminating between antibiotic-treated (TX group) and untreated pigs (CTRL group), with the further perspective of identifying the authentication tools for antibiotic-free pork supply chains. In particular, 41 samples of pig liver were subjected to a biphasic extraction to recover both the polar and the non-polar metabolites, and the 1H NMR spectroscopy analysis was performed on the two separate extracts. Unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate statistical analysis of 1H NMR spectra data in the range 0–9 ppm provided metabolomic fingerprinting useful for the discrimination of pig livers based on the antibiotic treatment to which they were exposed. Moreover, within the signature patterns, significant discriminating metabolites were identified among carbohydrates, choline and derivatives, amino acids and some lipid-class molecules. The encouraging findings of this exploratory study showed the feasibility of the untargeted metabolomic approach as a novel strategy in the authentication framework of pork supply chains and open a new horizon for a more in-depth investigation.

Fuel spills are a major source of contamination in terrestrial environments in Antarctica. Little is known of the effects of hydrocarbon contaminants in fuels on Antarctic terrestrial biota, and how these change as fuel ages within soil. In this study we investigate the sensitivity of juveniles of the endemic Antarctic nematode Plectus murrayi to diesel-spiked soil. Toxicity tests were conducted on soil elutriates, and changes in concentrations of hydrocarbons, polar compounds and PAHs were assessed as the spiked soil was artificially aged at 3 °C over a 45-week period, representing multiple summer seasons of fuel degradation. Nematodes were most sensitive to elutriates made from freshly spiked soils (LC50 419 μg/L TPH and 156 μg/L TPH-SG), with a subsequent decline in toxicity observed in the first 6 weeks of laboratory ageing (LC50 2945 μg/L TPH and 694 μg/L TPH-SG). Effects were still evident up to 45 weeks (lowest observed effect concentration 2123 μg/L TPH) despite hydrocarbons being depleted from soils with ageing (84 % loss) and elutriates becoming dominated by polar metabolites (95 % polar). Nematode sensitivity throughout the ageing period showed evidence of a relationship between LC50 and the proportions of the lighter carbon range fraction of TPH in elutriates, the F2 fraction (C10–14). This study is the first to estimate the sensitivity of Antarctic terrestrial fauna to diesel and provides novel data on the dynamics of fuel chemistry under Antarctic conditions and how this influences toxicity. Findings contribute to predicting ecological risk at existing diesel fuel spill sites in Antarctica, to the derivation of site-specific remediation targets, and to environmental guidelines to assess ecosystem health.

Summary: The goal of this study was to use GC‐MS and UHPLC‐UV‐VIS‐based metabolomic tools combined with chemometric analysis to discriminate between fruit with blackspot physiological disorder independently of storage condition (regular air or controlled atmosphere) and without blackspot (hereafter referred to as 'Control'). The study looked at polar metabolites, enzymes, fatty acids and oxidative stress markers. Fruit with high levels of lenticel damage are more likely to develop blackspot, which appears to be exacerbated by H2O2 and peroxidation of lipids. Lower catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO), superoxide dismutase (SOD) and phenylalanine‐ammonia lyase (PAL) activities together with lower levels of polyphenols (syringic acid, epigallocatechin, quercetin and vanillic) and fatty acids (linolenic, linoleic and palmitic acid), as well as ß‐sitosterol, methyl‐3‐hydroxybutyrate, glyceric acid, hydroxyl butanoic acid, L‐threonic acid, hydroxyl glutaric acid and xylose contributed to a higher prevalence of blackspot disorder in stored Hass avocados, which was accompanied by upregulation of soluble proteins. Finally, chemometric analysis revealed a positive relationship between blackspot disorder with gluconic, palmitoleic, galactose oxime, glyceric, hydroxyglutaric, MDA, syringic and oleic acid, as well as a negative relationship with catechin, epigallocatechin, SOD, CAT and tagatose. The study concluded that oxidative stress is the main driver of 'Blackspot' physiological disorder. [ABSTRACT FROM AUTHOR]

In aquatic environments the concurrent exposure of molluscs to microplastics (MPs) and estrogens is common, as these pollutants are frequently released by wastewater treatment plants into estuaries. Therefore, this study aimed to evaluate the independent and co-exposure impacts of polyethylene microplastics (PE-MPs) and estrogenic endocrine-disrupting chemicals (EEDCs) at environmentally relevant concentrations on polar metabolites and morphological parameters of the Sydney rock oyster. A seven-day acute exposure revealed no discernible differences in morphology; however, significant variations in polar metabolites were observed across oyster tissues. The altered metabolites were mostly amino acids, carbohydrates and intermediates of the Kreb's cycle. The perturbation of metabolites were tissue and sex-specific. All treatments generally showed an increase of metabolites relative to controls – a possible stimulatory and/or a potential hormetic response. The presence of MPs impeded the exposure of adsorbed and free EEDCs potentially due to the selective feeding behaviour of oysters to microplastics, favouring algae over similar-sized PE-MPs, and the formation of an eco/bio-corona involving faeces, pseudo-faeces, natural organic matter, and algae. [Display omitted] • Effects of microplastics and estrogens are sex and tissue specific. • Single and mixed exposure similarly affect overall metabolic status. • At environmentally realistic concentrations, contaminants exerted a stimulatory effect. • Exposure to low concentrations of both contaminants may present a hormetic response. [ABSTRACT FROM AUTHOR]

The metabolic profiling of a wide range of chemical classes relevant to understanding sarcopenia under conditions in which sample availability is limited, e.g., from mouse models, small muscles, or muscle biopsies, is desired. Several existing metabolomics platforms that include diverse classes of signaling lipids, energy metabolites, and amino acids and amines would be informative for suspected biochemical pathways involved in sarcopenia. The sample limitation requires an optimized sample preparation method with minimal losses during isolation and handling and maximal accuracy and reproducibility. Here, two developed sample preparation methods, BuOH-MTBE-Water (BMW) and BuOH-MTBE-More-Water (BMMW), were evaluated and compared with previously reported methods, Bligh-Dyer (BD) and BuOH-MTBE-Citrate (BMC), for their suitability for these classes. The most optimal extraction was found to be the BMMW method, with the highest extraction recovery of 63% for the signaling lipids and 81% for polar metabolites, and an acceptable matrix effect (close to 1.0) for all metabolites of interest. The BMMW method was applied on muscle tissues as small as 5 mg (dry weight) from the well-characterized, prematurely aging, DNA repair-deficient Ercc1∆/− mouse mutant exhibiting multiple–morbidities, including sarcopenia. We successfully detected 109 lipids and 62 polar targeted metabolites. We further investigated whether fast muscle tissue isolation is necessary for mouse sarcopenia studies. A muscle isolation procedure involving 15 min at room temperature revealed a subset of metabolites to be unstable; hence, fast sample isolation is critical, especially for more oxidative muscles. Therefore, BMMW and fast muscle tissue isolation are recommended for future sarcopenia studies. This research provides a sensitive sample preparation method for the simultaneous extraction of non-polar and polar metabolites from limited amounts of muscle tissue, supplies a stable mouse muscle tissue collection method, and methodologically supports future metabolomic mechanistic studies of sarcopenia. [ABSTRACT FROM AUTHOR]

Understanding the chemical nature of wine aroma demands accurate quantitative determinations of different odor-active compounds. Quantitative determinations of enolones (maltol, furaneol, homofuraneol, and sotolon) and vanillin derivatives (vanillin, methyl vanillate, ethyl vanillate, and acetovanillone) at low concentrations are complicated due to their high polarity. For this reason, this paper presents an improved and automated version for the accurate measure of these common trace wine polar compounds (enolones and vanillin derivatives). As a result, a faster and more user-friendly method with a reduction of organic solvents and resins was developed and validated. The optimization of some stages of the solid phase extraction (SPE) process, such as washing with an aqueous solution containing 1% NaHCO3 at pH 8, led to cleaner extracts and solved interference problems. Due to the polarity of these type of compounds, an optimization of the large volume injection was also carried out. Finally, a programmable temperature vaporization (PTV) quartz glass inlet liner without wool was used. The injector temperature was raised to 300 °C in addition to applying a pressure pulse of 180 kPa for 4 min. Matrix effects were solved by the use of adequate internal standards, such as ethyl maltol and 3′,4′-(methylenedioxy)acetophenone. Method figures of merit were highly satisfactory: good linearity (r2 > 0.98), precision (relative standard deviation, RSD < 10%), high recovery (RSD > 89%), and low detection limits ( 1) while homofuraneol and sotolon could also produce changes in their aroma perceptions (0.1 < OAV < 1).

In epidemiological studies, samples are often collected long before disease onset or outcome assessment. Understanding the long-term stability of biomarkers measured in these samples is crucial. We estimated within-person stability over 10 years of metabolites and metabolite features (n = 5938) in the Nurses' Health Study (NHS): the primary dataset included 1880 women with 1184 repeated samples donated 10 years apart while the secondary dataset included 1456 women with 488 repeated samples donated 10 years apart. We quantified plasma metabolomics using two liquid chromatography mass spectrometry platforms (lipids and polar metabolites) at the Broad Institute (Cambridge, MA, USA). Intra-class correlations (ICC) were used to estimate long-term (10 years) within-person stability of metabolites and were calculated as the proportion of the total variability (within-person + between-person) attributable to between-person variability. Within-person variability was estimated among participants who donated two blood samples approximately 10 years apart while between-person variability was estimated among all participants. In the primary dataset, the median ICC was 0.43 (1st quartile (Q1): 0.36; 3rd quartile (Q3): 0.50) among known metabolites and 0.41 (Q1: 0.34; Q3: 0.48) among unknown metabolite features. The three most stable metabolites were N6,N6-dimethyllysine (ICC = 0.82), dimethylguanidino valerate (ICC = 0.72), and N-acetylornithine (ICC = 0.72). The three least stable metabolites were palmitoylethanolamide (ICC = 0.05), ectoine (ICC = 0.09), and trimethylamine-N-oxide (ICC = 0.16). Results in the secondary dataset were similar (Spearman correlation = 0.87) to corresponding results in the primary dataset. Within-person stability over 10 years is reasonable for lipid, lipid-related, and polar metabolites, and varies by metabolite class. Additional studies are required to estimate within-person stability over 10 years of other metabolites groups. [ABSTRACT FROM AUTHOR]

Rapid diagnosis of diseases is one of the challenging areas in clinical research. From the analytical chemist's perspective, the main challenges are isolating the compounds from the bio-specimen and lengthy analysis times. In this regard, solid phase microextraction offers a platform to address the abovementioned challenges. Moreover, its sharp tip-thin film geometry, known as coated blade spray (CBS), can enhance the extraction and act as an ionization source in direct mass spectrometric analysis. In this study, a new CBS device specifically designed for polar analytes was prepared and optimized to determine urinary metabolites. For this purpose, polyacrylonitrile (PAN) was selected as a base polymer as it can be electrospun to form a nanofibrous structure, and it can be modified with weak ion exchange moieties to interact with polar analytes. Following the electrospinning of PAN, hydrolysis was optimized, and conditions leading to sufficient extraction enhancement without dissolving the polymer were obtained when probes were treated with 5.0 M of NaOH for 2.5 h. Using the coated blades prepared as explained, the evaluation of various extraction conditions showed that 5 min is sufficient for equilibrium extraction. In addition, the solution's ionic strength and pH significantly affect the extraction. Optimum sorption was obtained at no salt added and pH 7.0 conditions. The CBS-MS optimization showed that 10.0 µL of ACN/MeOH/H 2 O (40:40:20, v/v/v) with formic acid kept for 15 seconds on the blade before voltage application leads to the highest signal. The limits of quantification of the analytes are between 50 and 100 ng/mL. [Display omitted] • Hydrolyzed electrospun PAN can be used for extraction of polar metabolites. • New electrospun based extractive phase provides fast extraction kinetics. • The CBS-MS method is only 6 min and can be used as a fast diagnostic tool. [ABSTRACT FROM AUTHOR]

The choice of rootstock has a significant impact, not only on fruit growth and development, but also on avocado fruit quality and postharvest performance. The objective of this study was to evaluate and compare attributes related to the postharvest quality of Hass avocados from “Mexicola” and “Duke 7” rootstocks grown under similar conditions. This study included two harvests: early (23–26% dry matter) and middle (>26–30% dry matter) per season for the 2016/2017 and 2017/2018 seasons and two storage conditions (regular air (RA) at 5 °C and controlled atmosphere (CA) at 4 kPa O2 and 6 kPa CO2 at 5 °C) for 55 days. The results showed significant differences in firmness, color and vascular and flesh browning between storage conditions; in addition to these last three attributes, the rootstock played an important role. The fatty acid profile did not reveal significant differences between fruit from both rootstocks. Finally, the polar metabolite profiles revealed differences only for the storage condition, not associated to the rootstock, which could affect the postharvest performance of Hass avocado fruit. This study is one of the few available showing the interaction of rootstock/Hass cultivar on fruit quality and postharvest performance.

Harvested avocado fruits can be potentially exposed both to pre- and post-harvest factors that may influence the susceptibility of fruit to quality loss. Regular air (RA) storage consists of approximately 78% nitrogen, 21% oxygen, 0.3% carbon dioxide, and smaller amounts of some other gasses. A controlled atmosphere (CA) is a changed atmosphere that is used to delay respiration and reduce fungal and physiological deterioration, and it is an appropriate and practical technology for prolonging the shelf life and maintaining the quality of fruits on a large scale. An experiment was conducted by collecting fruits from two different avocado orchards (Bartolillo and Quilhuica) during two harvest seasons based on dry matter content and two subsequent years. The current study was designed to investigate the changes in polar metabolites, phenolic compounds, and fatty acids in the skin of Hass avocados stored under two distinct conditions. Bartolillo orchard fruits were mostly correlated to linoleic and oleic acid as important variables. For Quilhuica, fruits were mostly correlated with palmitoleic, palmitic, and oleic acids. Phenolic content increased at the beginning of storage and decreased at the end of storage for one orchard and contrarily for others, indicating that the result was dependent on the orchard and storage condition. Serine, glutaric acid, xylitol and D-mannitol were the polar metabolites that most correlated with fruits of the Quilhuica orchard while ß-sitosterol and gluconic were related to fruits of the Bartolillo orchard. The differences in fatty acids, polar metabolites and phenolics were dependent on orchard and storage conditions.

The antibacterial activity of methanolic extracts of eight Mediterranean brown algae belonging to the genera Cystoseira sensu lato, Dictyopteris and Padina was investigated. Extracts from Sargassaceae showed antibacterial activity. Specifically, Carpodesmia crinita and Carpodesmia brachycarpa extracts inhibited the growth of the Gram-positive strain Kocuria rhizophila whereas Cystoseira compressa and Carpodesmia amentacea extracts showed antibacterial activity against the Gram-positive strains, Kocuria rhizophila and Staphylococcus aureus. None of them inhibited the growth of the Gram-negative bacterium Escherichia coli. The extracts of Cystoseira sp. pl. and Treptacantha ballesterosii did not show any antibacterial activity. HPLC-MS analysis of the extracts allowed identification of polar compounds. Taxa of the genus Cystoseira sensu lato showed a higher number of compounds than Dictyopteris polypodioides and Padina pavonica. Some differences found among and within taxa can be attributed to the different depths where these algae live, to thallus morphology and herbivore pressure. Particularly interesting were the phenol compounds, which showed differences among the genera but also within the genus Cystoseira sensu lato. Noteworthy, the almost exclusive presence of the fukiik acid isomer and of bromo-phloroglucinol in Carpodesmia amentacea, and the presence of exifone, able to bind iron metal ions, in Carpodesmia amentacea, Cystoseira compressa and Treptacantha ballesterosii. [ABSTRACT FROM AUTHOR]

Recently, wheat has attracted attention as a functional food, rather than a simple dietary energy source. Accordingly, whole-grain intake increases with an understanding of bioactive phytochemicals in bran. The development of colored wheat has drawn more attention to the value of bran owing to its nutritional quality, as well as the antioxidant properties of the colorant. The present 1H NMR-based chemometric study evaluated the compositional improvement of radiation-induced mutants in purple wheat by focusing on the predominant metabolites with high polarity. A total of 33 metabolites, including three choline derivatives, three sugar alcohols, four sugars, 13 amino acids, eight organic acids, and two nucleosides, were identified throughout the 1H NMR spectra, and quantification data were obtained for the identified metabolites via peak shape-based quantification. Principal component and hierarchical cluster analyses were conducted for performing multivariate analyses. The colored original wheat was found to exhibit improvements compared to yellow wheat in terms of the contents of primary metabolites, thus highlighting the importance of conducting investigations of polar metabolites. The chemometrics studies further revealed mutant lines with a compositional enhancement for metabolites, including lysine, proline, acetate, and glycerol.

Accumulation of anthocyanins in detached leaves and in excised stems of Kalanchoë blossfeldiana kept under natural light conditions in the presence or absence of methyl jasmonate (JA-Me) was investigated. When the abaxial surface of detached leaves was held lower than the adaxial surface (the normal or natural position) under natural light conditions, anthocyanins were not accumulated on the abaxial side of the leaves. In contrast, when the adaxial surface of detached leaves was held lower than the abaxial surface (inverted position), anthocyanins were highly accumulated on the abaxial side of the leaves. These phenomena were independent of the growth stage of K. blossfeldiana as well as photoperiod. Application of JA-Me in lanolin paste significantly inhibited anthocyanin accumulation induced on the abaxial side of detached leaves held in an inverted position in a dose-dependent manner. Anthocyanin accumulation in the excised stem in response to natural light was also significantly inhibited by JA-Me in lanolin paste. Possible mechanisms of anthocyanin accumulation on the abaxial side of detached K. blossfeldiana leaves held in an inverted position under natural light conditions and the inhibitory effect of JA-Me on this process are described. The accompanying changes in the content of primary metabolites and histological analyses were also described.

The metabolic profiling of a wide range of chemical classes relevant to understanding sarcopenia under conditions in which sample availability is limited, e.g., from mouse models, small muscles, or muscle biopsies, is desired. Several existing metabolomics platforms that include diverse classes of signaling lipids, energy metabolites, and amino acids and amines would be informative for suspected biochemical pathways involved in sarcopenia. The sample limitation requires an optimized sample preparation method with minimal losses during isolation and handling and maximal accuracy and reproducibility. Here, two developed sample preparation methods, BuOH-MTBE-Water (BMW) and BuOH-MTBE-More-Water (BMMW), were evaluated and compared with previously reported methods, Bligh-Dyer (BD) and BuOH-MTBE-Citrate (BMC), for their suitability for these classes. The most optimal extraction was found to be the BMMW method, with the highest extraction recovery of 63% for the signaling lipids and 81% for polar metabolites, and an acceptable matrix effect (close to 1.0) for all metabolites of interest. The BMMW method was applied on muscle tissues as small as 5 mg (dry weight) from the well-characterized, prematurely aging, DNA repair-deficient Ercc1∆/− mouse mutant exhibiting multiple–morbidities, including sarcopenia. We successfully detected 109 lipids and 62 polar targeted metabolites. We further investigated whether fast muscle tissue isolation is necessary for mouse sarcopenia studies. A muscle isolation procedure involving 15 min at room temperature revealed a subset of metabolites to be unstable; hence, fast sample isolation is critical, especially for more oxidative muscles. Therefore, BMMW and fast muscle tissue isolation are recommended for future sarcopenia studies. This research provides a sensitive sample preparation method for the simultaneous extraction of non-polar and polar metabolites from limited amounts of muscle tissue, supplies a stable mouse muscle tissue collection method, and methodologically supports future metabolomic mechanistic studies of sarcopenia.

• The metabolic phenotyping of the polar metabolites excreted in urine is challenging. • Urinalysis using HILIC is currently the method of choice for polar metabolites. • Ion mobility-enabled analysis (HILIC-IM-MS) significantly increased peak detection. • A cyclic IM-MS instrument gave enhanced peak detection compared to a linear IM-MS configuration. • HILIC-IM-MS improved both coverage of the polar urinary metabolome and MS data. The use of HILIC-based separations for the analysis of polar metabolites in metabolic phenotyping studies is well established. Here, we demonstrate the increased coverage of the polar metabolome obtained by travelling wave (TW) ion mobility (IM) instruments combined with HILIC and mass spectrometry (MS) for metabotyping rat and mouse urine samples. Profiling was performed using either a linear TW IM-MS based instrument with a path length of 40 cm or an instrument with a cyclic travelling wave analyser (cIM) with a path length of 95 cm. Due to the added resolution afforded by using both the linear and cyclic IM geometries with MS detection (IM-MS) significant increases in feature count (m/z -t R pairs) were generally obtained compared to HILIC-MS alone. In addition, the use of both linear and cyclic IM-MS improved the quality of the mass spectra obtained as a result of the separation of co-eluting analytes. As would be expected from the increased path length of the cyclic IM-MS instrument compared to the linear device, the largest gains in feature detection were obtained for the HILIC-cIM-MS combination. By increasing the resolution of coeluting components, the cyclic IM-MS instrumentation also provided the largest improvement in the quality of the mass spectral data obtained. When applied to mouse urines obtained from both control and gefitinib-dosed mice, time-related changes were detected in those obtained from the treated animals that were not seen in the controls. Polar metabolites affected by drug administration included, but were not limited to, hypoxanthine, 1,3-dimethyluracil and acetylcarnitine. The changes seen in the relative concentrations of these endogenous metabolites appeared to be related to drug concentrations in the plasma and urine suggesting a pharmacometabodynamic link. [ABSTRACT FROM AUTHOR]

In epidemiological studies, samples are often collected long before disease onset or outcome assessment. Understanding the long-term stability of biomarkers measured in these samples is crucial. We estimated within-person stability over 10 years of metabolites and metabolite features (n = 5938) in the Nurses’ Health Study (NHS): the primary dataset included 1880 women with 1184 repeated samples donated 10 years apart while the secondary dataset included 1456 women with 488 repeated samples donated 10 years apart. We quantified plasma metabolomics using two liquid chromatography mass spectrometry platforms (lipids and polar metabolites) at the Broad Institute (Cambridge, MA, USA). Intra-class correlations (ICC) were used to estimate long-term (10 years) within-person stability of metabolites and were calculated as the proportion of the total variability (within-person + between-person) attributable to between-person variability. Within-person variability was estimated among participants who donated two blood samples approximately 10 years apart while between-person variability was estimated among all participants. In the primary dataset, the median ICC was 0.43 (1st quartile (Q1): 0.36; 3rd quartile (Q3): 0.50) among known metabolites and 0.41 (Q1: 0.34; Q3: 0.48) among unknown metabolite features. The three most stable metabolites were N6,N6-dimethyllysine (ICC = 0.82), dimethylguanidino valerate (ICC = 0.72), and N-acetylornithine (ICC = 0.72). The three least stable metabolites were palmitoylethanolamide (ICC = 0.05), ectoine (ICC = 0.09), and trimethylamine-N-oxide (ICC = 0.16). Results in the secondary dataset were similar (Spearman correlation = 0.87) to corresponding results in the primary dataset. Within-person stability over 10 years is reasonable for lipid, lipid-related, and polar metabolites, and varies by metabolite class. Additional studies are required to estimate within-person stability over 10 years of other metabolites groups.

Polar hydrophilic metabolites have been identified as important actors in many biochemical pathways. Despite continuous improvement and refinement of hydrophilic interaction liquid chromatography (HILIC) platforms, its application in global polar metabolomics has been underutilized. In this study, we aimed to systematically evaluate polar stationary phases for untargeted metabolomics by using HILIC columns (neutral and zwitterionic) that have been exploited widely in targeted approaches. To do so, high-resolution mass spectrometry was applied to thoroughly investigate selectivity, repeatability and matrix effect at three pH conditions for 9 classes of polar compounds using 54 authentic standards and plasma matrix. The column performance for utilization in untargeted metabolomics was assessed using plasma samples with diverse phenotypes. Our results indicate that the ZIC-c HILIC column operated at neutral pH exhibited several advantages, including superior performance for different classes of compounds, better isomer separation, repeatability and high metabolic coverage. Regardless of the column type, the retention of inorganic ions in plasma leads to extensive adduct formation and co-elution with analytes, which results in ion-suppression as part of the overall plasma matrix effect. In ZIC-c HILIC, the sodium chloride ion effect was particularly observed for amino acids and amine classes. Successful performance of HILIC for separation of plasma samples with different phenotypes highlights this mode of separation as a valuable approach in global profiling of plasma sample and discovering the metabolic changes associated with health and disease.

Metabolomics is a powerful approach in evaluating the health status of organisms in ecotoxicological studies. However, metabolomics data reflect metabolic variations that are attributable to factors intrinsic to the environment and organism, and it is thus crucial to accurately evaluate the metabolome of the tissue/organ examined when it is exposed to no stressor. The metabolomes of the liver and gills of wild golden grey mullet (Chelon auratus) from a reference area were analyzed and compared by proton nuclear magnetic resonance (1H NMR)-based metabolomics. Both organs were characterized by amino acids, carbohydrates, osmolytes, nucleosides and their derivatives, and miscellaneous metabolites. However, similarities and differences were revealed in their metabolite profile and related to organ-specific functions. Taurine was predominant in both organs due to its involvement in osmoregulation in gills, and detoxification and antioxidant protective processes in liver. Environmental exposure to mercury (Hg) triggered multiple and often differential metabolic alterations in fish organs. Disturbances in ion-osmoregulatory processes were highlighted in the gills, whereas differential impairments between fish organs were pointed out in energy-producing metabolic pathways, protein catabolism, membrane stabilization processes, and antioxidant defense system, reflecting the induction of organ-specific adaptive and defensive strategies. Overall, a strict correlation between metabolites and organ-specific functions of fish gills and liver were discerned in this study, as well as organ-specific cytotoxicity mechanisms of Hg in fish.

The potential toxicity to human and aquatic receptors of petroleum fuel biodegradation metabolites (oxygen‐containing organic compounds [OCOCs]) in groundwater has been investigated as part of a multi‐year research program. Whole mixtures collected from locations upgradient and downgradient of multiple fuel release sites were tested using: 1) in vitro screening assays for human genotoxicity (the gamma‐H2AX assay) and estrogenic effects (estrogen receptor transcriptional activation assay), and 2) chronic aquatic toxicity tests in 3 species (Ceriodaphnia dubia, Raphidocelis subcapitata, and Pimephales promelas). In vitro screening assay results demonstrated that the mixtures did not cause genotoxic or estrogenic effects. No OCOC‐related aquatic toxicity was observed and when aquatic toxicity did occur, upgradient samples typically had the same response as samples downgradient of the release, indicating that background water quality was impacting the results. This information provides additional support for previous work that focused on the individual compounds and, taken together, indicates that OCOCs from petroleum degradation at fuel release sites are unlikely to cause toxicity to human or freshwater receptors at the concentrations present. Environ Toxicol Chem 2020;39:1634–1645. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. [ABSTRACT FROM AUTHOR]

Abstract Background Recent advances in the mapping of biochemical traits have been reported in Lolium perenne. Although the mapped traits, including individual sugars and fatty acids, contribute greatly towards ruminant productivity, organic acids and amino acids have been largely understudied despite their influence on the ruminal microbiome. Results In this study, we used a targeted gas-chromatography mass spectrometry (GC-MS) approach to profile the levels of 25 polar metabolites from different classes (sugars, amino acids, phenolic acids, organic acids and other nitrogen-containing compounds) present in a L. perenne F2 population consisting of 325 individuals. A quantitative trait (QTL) mapping approach was applied and successfully identified QTLs regulating seven of those polar metabolites (L-serine, L-leucine, glucose, fructose, myo-inositol, citric acid and 2, 3-hydroxypropanoic acid).Two QTL mapping approaches were carried out using SNP markers on about half of the population only and an imputation approach using SNP and DArT markers on the entire population. The imputation approach confirmed the four QTLs found in the SNP-only analysis and identified a further seven QTLs. Conclusions These results highlight the potential of utilising molecular assisted breeding in perennial ryegrass to modulate a range of biochemical quality traits with downstream effects in livestock productivity and ruminal digestion.

Hydrocarbon polar metabolites are gaining interest from industry and the remediation community due to their ubiquity and uncertainty around their toxicity. In this study, we used headspace-gas chromatography/mass spectrometry (HS-GC/MS) to characterize polar metabolites present in elutriates derived from uncontaminated, freshly hydrocarbon contaminated and partially remediated Antarctic soils. Elutriates represent the bioavailable fraction and may be used as a proxy for leachate runoff in environmental risk assessments. Control and contaminated soil elutriates were analysed for the presence of 12 aldehydes and two ketones, which cover a broad spectrum of metabolites, ranging from nC 2 – nC 12 carbon chain length. A total of nine aldehydes were detected in the soil elutriates. Types of aldehydes present in uncontaminated and hydrocarbon contaminated elutriates were similar. Among the polar metabolites measured in elutriates, acetaldehyde was most abundant in partially remediated soils. Microtox assays were used to determine the potential toxicity of elutriates. In addition, three aldehydes that were present at the highest concentrations in the contaminated and partially remediated soil elutriates (acetaldehyde, octanal and undecanal) were tested as single compounds. Contaminated soil elutriates tested were found to be toxic, with partially remediated elutriates less toxic than freshly contaminated elutriates. None of the three aldehydes tested separately were toxic at levels at which they were measured in elutriates. We infer that high levels of acetaldehyde in partially remediated soil due to hydrocarbon degradation highlight the potential of this metabolite as a useful chemical marker for hydrocarbon degradation under certain conditions. Microtox was sensitive to metabolites and provided a useful initial screening tool for elutriates. Unlabelled Image • Polar metabolites have a higher solubility than their parent hydrocarbons and are potentially bioavailable. • Nine aldehydes ranging from nC 2 -nC 12 were found in hydrocarbon contaminated soil elutriates. • Aldehydes could be potential chemical markers for hydrocarbon degradation. [ABSTRACT FROM AUTHOR]

Background and objectives: The changes in polar metabolites and volatiles in sesame seeds during roasting were analyzed to provide new information for the formation mechanism of sesame oil aroma. Findings: The results showed that the typical sesame aroma components, heterocyclic compounds, increased with the extension of the roasting time and reached 43.78% at 12 min. However, the proportion of aliphatics and terpenoids decreased with the roasting time. In addition, the main polar metabolites in sesame seeds, free amino acids and monosaccharides, continually decreased from 497.14 μg/g and 268.85 μg/g in raw seeds to 144.59 μg/g and 7.52 μg/g in 12‐min roasted seeds, respectively. Conclusions: Based on the changes in specific polar metabolites and volatile compounds, it could be inferred that pyrazines mainly generated through Maillard reaction with Ala and glucose as the possible primary substrates. While S‐heterocycles might be mainly formed through the thermal degradation of proteins and subsequent reactions. Significance and novelty: The results of this study provide important information and new details for the aroma formation process of sesame seeds during roasting and would provide a reference for the study and optimization of sesame roasting and oil extraction processes, as well as the development of sesame seed products. In addition, relevant information can also provide clues for the artificial synthesis of sesame seed aroma components. [ABSTRACT FROM AUTHOR]

Extraction of polar endogenous compounds remains an important issue in bioanalysis although different techniques have been evaluated. Among them, electromembrane extraction (EME) is a relevant approach but supported liquid membranes (SLMs) dedicated to polar molecules are still lacking. In this study 22 organic solvents were evaluated as SLMs on a set of 45 polar basic metabolites (log P from −5.7 to 1.5) from various biochemical families. To investigate a large variety of organic solvents, a parallel electromembrane extraction device was used and a constant current approach was applied to circumvent the heterogeneous conductivities of the different SLMs. Among the tested organic solvents, 2-nitrophenyl pentyl ether (NPPE) appeared the most efficient SLM with the extraction of a large variety of polar cationic metabolites, high extraction yields, and low extraction variabilities. The applied current and the composition of the acceptor and donor solutions were also evaluated and 300 μA per well and acetic acid 1% (v/v), both as acceptor and donor compartments, were the most efficient conditions. The new SLM and the optimized experimental parameters were successfully applied to the extraction of precipitated plasma samples. Although the extraction recovery decreased for most compounds in the biological matrix, process efficiency (PE) up to 90% and low extraction variability (RSD between 2 and 18%) were obtained for several very polar compounds such as choline or acetylcholine, emphasizing the potential of EME for polar compounds. [ABSTRACT FROM AUTHOR]

A simple and effective method was developed for analyzing dinotefuran and its three metabolites (MNG, UF, and DN) in plum using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Due to the polarity and high water miscibility, dinotefuran and some of its metabolites (especially DN) have some limitations to be extracted with acetonitrile and salt following the “QuEChERS” sample preparation methodology. Alternatively, the samples were extracted with methanol, and purified with dispersive-solid phase extraction procedure (d-SPE) using primary secondary amine (PSA) and C 18 sorbents after filtration, and mass up. Due to the suppression effect originated from plum matrix, matrix-matched calibration curves, which provided good linearity with coefficient of determination ( R 2 ) ≥ 0.998, were used for quantification of all analytes. Blank plum samples fortified with 2 spiking levels (10 × LOQ and 50 × LOQ) yielded satisfactory recoveries for all tested analytes in the range of 83.01 to 110.18% with relative standard deviation (RSD) ≤ 8.91. The method was successfully applied to field-incurred plum samples and dinotefuran and all metabolites were positively detected and quantified. In conclusion, we suggest that the method can be expanded to polar compounds having solvent and partitioning problems in any of the versions of QuEChERS. [ABSTRACT FROM AUTHOR]

A HILIC-UHPLC–MS/MS untargeted urinary metabonomic method combined with quantitative analysis of five potential polar biomarkers in rat urine was developed and validated, to further understand the anti-osteoporosis effect of Gushudan(GSD) and its mechanism on prednisolone-induced osteoporosis(OP) rats in this study. The metabolites were separated and identified on Waters BEH HILIC (2.1 mm × 100 mm, 1.7 μm) column using the Waters ACQUITY™ ultra performance liquid chromatography system (Waters Corporation, Milford, USA) coupled with a Micromass Quattro Micro™ API mass spectrometer (Waters Corp, Milford, MA, USA). Principal component analysis (PCA) was used to identify potential biomarkers. Primary potential polar biomarkers including creatinine, taurine, betaine, hypoxanthine and cytosine, which were related to energy metabolism, lipid metabolism and amino acid metabolism, were found in the untargeted metabonomic research. Moreover, these targeted biomarkers were further separated and quantified in multiple-reaction monitoring (MRM) with positive ionization mode, using tinidazole as internal standard (I.S.). Good linearities (r > 0.99) were obtained for all the analytes with the low limit of quantification from 1.00 to 12.8 μg/mL. The relative standard deviation (RSD) of the intra-day and inter-day precisions were within 15.0% and the accuracy ranged from −14.3% to 13.5%. The recovery was more than 85.0%. And the validated method was successfully applied to investigate the urine samples of the control group, prednisolone-induced osteoporosis model group and Gushudan-treatment group in rats. Compared to the control group, the level of creatinine, taurine, betaine, hypoxanthine and cytosine in the model group revealed a significant decrease trend (p < 0.05), while the Gushudan-treatment group showed no statistically differences by an independent sample t -test. This paper provided a better understanding of the therapeutic effect and mechanism of GSD on prednisolone-induced osteoporosis rats. [ABSTRACT FROM AUTHOR]

Background: Recent advances in the mapping of biochemical traits have been reported in Lolium perenne. Although the mapped traits, including individual sugars and fatty acids, contribute greatly towards ruminant productivity, organic acids and amino acids have been largely understudied despite their influence on the ruminal microbiome. Results: In this study, we used a targeted gas-chromatography mass spectrometry (GC-MS) approach to profile the levels of 25 polar metabolites from different classes (sugars, amino acids, phenolic acids, organic acids and other nitrogen-containing compounds) present in a L. perenne F2 population consisting of 325 individuals. A quantitative trait (QTL) mapping approach was applied and successfully identified QTLs regulating seven of those polar metabolites (L-serine, L-leucine, glucose, fructose, myo-inositol, citric acid and 2, 3-hydroxypropanoic acid). Two QTL mapping approaches were carried out using SNP markers on about half of the population only and an imputation approach using SNP and DArT markers on the entire population. The imputation approach confirmed the four QTLs found in the SNP-only analysis and identified a further seven QTLs. Conclusions: These results highlight the potential of utilising molecular assisted breeding in perennial ryegrass to modulate a range of biochemical quality traits with downstream effects in livestock productivity and ruminal digestion. [ABSTRACT FROM AUTHOR]

A hydrophilic interaction liquid chromatography (HILIC)−ultra high-pressure liquid chromatography (UHPLC) coupled with tandem mass spectrometry (MS/MS) method was developed and applied to profile metabolite changes in human Huh-7 cells exposed to the potent aryl hydrocarbon receptor (AHR) ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Comparisons of sensitivity (limit of detection as low as 0.01 µM) and reproducibility (84% of compounds had an interday relative standard deviation (RSD) less than 10.0%; 83% of compounds had an intraday RSD less than 15.0%) were assessed for all the metabolites. The exposure of Huh-7 cells to the hepatotoxic carcinogen TCDD at low doses (1 nM and 10 nM for 4 h and 24 h, respectively) was reflected by the disturbance of amino acid metabolism, energy metabolism (glycolysis, TCA cycle), and nucleic acid metabolism. TCDD caused a significant decrease in amino acids such as serine, alanine, and proline while promoting an increase in arginine levels with 24 h treatment. Energy metabolism intermediates such as phosphoenolpyruvate and acetyl−CoA and nucleosides such as UMP, XMP, and CMP were also markedly decreased. These results support the application of HILIC−UHPLC−MS/MS for robust and reliable analysis of the cellular response to environmentally relevant toxicants at lower doses.

Fuel spills are a major source of contamination in terrestrial environments in Antarctica. Little is known of the effects of hydrocarbon contaminants in fuels on Antarctic terrestrial biota, and how these change as fuel ages within soil. In this study we investigate the sensitivity of juveniles of the endemic Antarctic nematode Plectus murrayi to diesel-spiked soil. Toxicity tests were conducted on soil elutriates, and changes in concentrations of hydrocarbons, polar compounds and PAHs were assessed as the spiked soil was artificially aged at 3 °C over a 45-week period, representing multiple summer seasons of fuel degradation. Nematodes were most sensitive to elutriates made from freshly spiked soils (LC50 419 μg/L TPH and 156 μg/L TPH-SG), with a subsequent decline in toxicity observed in the first 6 weeks of laboratory ageing (LC50 2945 μg/L TPH and 694 μg/L TPH-SG). Effects were still evident up to 45 weeks (lowest observed effect concentration 2123 μg/L TPH) despite hydrocarbons being depleted from soils with ageing (84 % loss) and elutriates becoming dominated by polar metabolites (95 % polar). Nematode sensitivity throughout the ageing period showed evidence of a relationship between LC50 and the proportions of the lighter carbon range fraction of TPH in elutriates, the F2 fraction (C 10–14). This study is the first to estimate the sensitivity of Antarctic terrestrial fauna to diesel and provides novel data on the dynamics of fuel chemistry under Antarctic conditions and how this influences toxicity. Findings contribute to predicting ecological risk at existing diesel fuel spill sites in Antarctica, to the derivation of site-specific remediation targets, and to environmental guidelines to assess ecosystem health. [Display omitted] • First estimates of sensitivity of Antarctic terrestrial fauna to diesel fuel. • Fresh spiked soil elutriate most toxic, with toxicity persisting during ageing. • Hydrocarbons depleted in soil with ageing and polar metabolites dominated elutriates. • Nematode sensitivity correlated with F2 fraction (C 10–14) of TPH in elutriate. • Critical data for the prediction of ecological risk of Antarctic diesel fuel spills. [ABSTRACT FROM AUTHOR]