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Advances in metabolomics analysis and data treatment increase the knowledge of complex biological systems. One of the most used methodologies is gas chromatography-mass spectrometry (GC-MS) due to its robustness, high separation efficiency, and reliable peak identification through curated databases. However, methodologies are not standardized, and the derivatization steps in GC-MS can introduce experimental errors and take considerable time, exposing the samples to degradation. Here, we propose the injection-port derivatization (IPD) methodology to increase the throughput in plasma metabolomics analysis by GC-MS. The IPD method was evaluated and optimized for different families of metabolites (organic acids, amino acids, fatty acids, sugars, sugar phosphates, etc.) in terms of residence time, injection-port temperature, and sample/derivatization reagent ratio. Finally, the method's usefulness was validated in a study consisting of a cohort of obese patients with or without nonalcoholic steatohepatitis. Our results show a fast, reproducible, precise, and reliable method for the analysis of biological samples by GC-MS. Raw data are publicly available at MetaboLights with Study Identifier MTBLS5151.

Electro-Fenton (EF) and peroxyelectrocoagulation (PEC) processes were investigated to mineralize 10 mg L−1 erythromycin from ultrapure water, evaluating the influence of the anode material (BDD and Fe), current density (janode) (5 mA cm−2 and 10 mA cm−2), oxygen flowrate injected to the cathode (0.8 L min−1 O2 and 2.0 L min−1 O2) and pH (2.8, 5.0 and 7.0) on the process efficiency and the electricity costs. 70% mineralization was reached after applying 0.32 A h L−1 under the best operational conditions: PEC treatment at 5 mA cm−2, 2.0 L min−1 O2 and pH 2.8. The electricity consumption of the electrochemical cell under these conditions was approximately 0.3 kWh m−3. Early-stage intermediates produced from erythromycin degradation were identified and quantified throughout the treatment and a potential erythromycin degradation pathway was proposed. The most appropriate operational conditions tested with synthetic solutions were applied to treat a real effluent from the tertiary treatment of an urban wastewater treatment plant. All emerging compounds listed in the EU Decision 2018/840 (Watch List 2018) were determined before and after the PEC treatment. All listed pollutants were degraded below their quantification limit, except estrone and 17-α-ethinylestradiol which were 99% removed from water. Electricity consumption of the electrochemical cell was 0.4 kWh m−3. Whilst awaiting future results that demonstrate the innocuity of the generated byproducts, the results of this investigation (high removal yields for emerging pollutants together with the low electricity consumption of the cell) indicate the promising high potential of PEC treatment as a water treatment/remediation/regeneration technology.

Hepatic biopsy is the gold standard for staging nonalcoholic fatty liver disease (NAFLD). Unfortunately, accessing the liver is invasive, requires a multidisciplinary team and is too expensive to be conducted on large segments of the population. NAFLD starts quietly and can progress until liver damage is irreversible. Given this complex situation, the search for noninvasive alternatives is clinically important. A hallmark of NAFLD progression is the dysregulation in lipid metabolism. In this context, recent advances in the area of machine learning have increased the interest in evaluating whether multi-omics data analysis performed on peripheral blood can enhance human interpretation. In the present review, we show how the use of machine learning can identify sets of lipids as predictive biomarkers of NAFLD progression. This approach could potentially help clinicians to improve the diagnosis accuracy and predict the future risk of the disease. While NAFLD has no effective treatment yet, the key to slowing the progression of the disease may lie in predictive robust biomarkers. Hence, to detect this disease as soon as possible, the use of computational science can help us to make a more accurate and reliable diagnosis. We aimed to provide a general overview for all readers interested in implementing these methods.

Nowadays, there is a strong interest in analytical approaches for assessing organic farming practices. Here, we propose that oxylipins, a group of oxidised metabolites derived from various polyunsaturated fatty acids, could be promising biomarkers for organic milk assessment because their biosynthesis is modulated by both precursor fatty acid availability and physiological or pathological status. Thus, we determined 31 fatty acids, 53 triacylglycerols and 37 oxylipins in one hundred commercial UHT milks by chromatographic methods coupled to mass spectrometry. Of these, 52 milks were conventional (34 whole milk, 11 semi-skimmed milk and 7 skimmed milk) and 48 were organic (31 whole milk, 11 semi-skimmed milk and 6 skimmed milk). Several oxylipins (8-HEPE, 5-HEPE, 11-HEPE, 9-HEPE, 18-HEPE, 9-HOTrE, 13-HOTrE, 12,13-DiHODE and 15,16-DiHODE) could distinguish between organic and conventional milks. Within these oxylipins, arachidonic and linoleic acid derived do not correlate with their fatty acid precursors; therefore these oxylipins could be promising as not only diet-dependent biomarkers for organic milk assessment.

Here we analysed the content of primary and secondary metabolites in nine types of industrially processed fibres derived from the juice industry. Specifically, we examined fibre from: apple, peach, and pear, as non-citrus fruits; the peel and flesh of orange and tangerine, and lemon flesh, as citrus fruits; and carrot, as vegetable. Regarding primary metabolites, the sugar content ranged from 21.6 mg/g in lemon to 290 mg/g in orange peel and lower mass organic acid content ranged from 25.0 mg/g in pear to 250 mg/g in lemon. The content of fatty acids were constant during fibre processing, ranging from 0.5 to 1.46%. Furthermore, the fatty acid profile was not affect for the processing. Concerning secondary metabolites, industrial processing did not decrease the sterols content, which ranged from 0.51 to 1.66 μg/g. Regarding carotenoids, of note was the presence of epoxycarotenoids, which may reflect the quality of the industrial process, thus giving added value to the by-product.

Colostrum and milk are the first nutrient sources for newborn piglets. In addition, n-3 fatty acids (FAs) and their oxygenated derivatives (oxylipins) have the capacity to modulate immune components. The aim of the current study was to include a fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in sow diets to promote an increase of anti-inflammatory molecules in colostrum and milk to benefit piglets. Thirty-six sows were randomly assigned from insemination to the end of lactation to either a control diet with animal fat (15 g/kg in gestation and 30 g/kg in lactation) or an n-3 diet in which animal fat was totally (gestation) or half (lactation) replaced by an equivalent amount of solid fish oil. Performance of sows and piglets was monitored during the study. Colostrum and milk samples were obtained after the birth of the first piglet and at weaning, respectively. From all samples (n = 18 per treatment), FAs were quantified by gas chromatography and immunoglobulins and cytokines by ELISA. Three samples per treatment were randomly selected to analyse oxylipin composition by liquid chromatography-tandem mass spectrometry. In colostrum and in milk, the n-3 FA (P = 0.020 and P < 0.001), particularly EPA (P < 0.001 and P < 0.001) and DHA (P < 0.001 and P < 0.001), and also their oxygenated derivatives were increased in samples from sows fed n-3 diet. Fish oil had no effect on immunoglobulin concentrations, but reduced tumour necrosis factor α (TNFα) (P = 0.011) and a tendency to reduce interleukin 10 (IL10) (P = 0.059) were observed in milk. In conclusion, fish oil in sow diets increased n-3 FA, particularly EPA and DHA, and their oxygenated derivatives in colostrum and milk, reducing TNFα and IL10 in milk. info:eu-repo/semantics/publishedVersion

Polyamines are involved in the regulation of many cellular functions and are promising biomarkers of numerous physiological conditions. Since the concentrations of these compounds in biological fluids are low, sample extraction is one of the most critical steps of their analysis. Here, we developed a comprehensive, sensitive, robust, and high-throughput LC-MS/MS stable-isotope dilution method for the simultaneous determination of 19 metabolites related to polyamine metabolism, including polyamines, acetylated and diacetylated polyamines, precursors, and catabolites from liquid biopsies. The sample extraction was optimized to remove interfering compounds and to reduce matrix effects, thus being useful for large clinical studies. The method consists of two-step liquid-liquid extraction with a Folch extraction and ethyl acetate partitioning combined with dansyl chloride derivatization. The developed method was applied to a small gender-related trial concerning human serum and urine samples from 40 obese subjects. Sex differences were found for cadaverine, putrescine, 1,3-diaminopropane, &gamma, aminobutyric acid, N8-acetylspermidine, and N-acetylcadaverine in urine, N1-acetylspermine in serum, and spermine in both serum and urine. The results demonstrate that the developed method can be used to analyze biological samples for the study of polyamine metabolism and its association with human diseases.

Here we authenticated single-varietal peach purees and pear juices on the basis of primary metabolite and phenolic compound analysis by Proton Nuclear Magnetic Resonance (1H-NMR) and Ultra Performance Liquid Chromatography coupled to Photodiode Array and Tandem Mass Spectrometry (UPLC-PDA-MS/MS), respectively. After suitable preprocessing, the 1H-NMR and chromatographic data were evaluated by principal component analysis (PCA). The PCA combining data from primary metabolites and phenolic compounds allowed the separation of the clusters in all cases, allowing discrimination of processed and unprocessed peach purees, both separately and pooled. The PCA of primary metabolites allowed the cluster separation of purees of distinct peach varieties but not between processed and non-processed purees. The PCA of phenolic compounds allowed better cluster separation than of primary metabolites. For pear juices, both PCA approaches allowed satisfactory discrimination of Alejandrina, Conference, and Blanquilla cultivars. These approaches may help to better control cultivar authenticity in fruit products. It could therefore contribute to the development of a process to achieve products characterized by a quality characteristic of a given cultivar. This research was funded by the Catalan Government, grant number [2017 SGR 828].

Objective: Polyamines are naturally occurring cationic molecules present in all living cells. Dysregulation of circulating polyamines has been reported in several conditions, but little is known about the levels of serum polyamines in chronic metabolic disorders such as type 2 diabetes (T2D). Therefore, the aim of this study was to evaluate the polyamine-related metabolome in a cohort of metabolic syndrome individuals with and without T2D. Design and methods: This was a nested case–control study within the PREDIMED-Plus trial that included 44 patients with T2D and 70 patients without T2D. We measured serum levels of arginine, ornithine, polyamines, and acetyl polyamines with an ultra-high performance liquid chromatography tandem mass spectrometry platform. Results: Our results showed that serum putrescine, directly generated from ornithine by the catalytic action of the biosynthetic enzyme ornithine decarboxylase, was significantly elevated in patients with T2D compared to those without T2D, and that it significantly correlated with the levels of glycosylated hemoglobin (HbA1c). Correlation analysis revealed a significantly positive association between fasting insulin levels and spermine. Multiple logistic regression analysis (adjusted for age, gender and body weight index) revealed that serum putrescine and spermine levels were associated with a higher risk of T2D. Conclusions: Our study suggests that polyamine metabolism is dysregulated in T2D, and that serum levels of putrescine and spermine are associated with glycemic control and circulating insulin levels, respectively.

Research is increasingly focused on identifying alternatives to the use of chemicals for the management of pests in stored grains. In this context, we tested the insecticidal and antifungal activities of methanolic extracts of Artemisia absinthium L. (Asteraceae), and two Cupressaceae species, namely Juniperus phoenicea L. and Tetraclinis articulata (Vahl) Mast., collected in the north of Algeria, against Sitophilus oryzae L. (Coleoptera, Curculionidea)—a pest of primary infestation of stored grains—and Fusarium sp., mycotoxigenic fungi. The extracts were analyzed by UPLC-PDA-MS. The extraction yield was in the range of 11–21 %. UPLC chromatographic data showed the presence of several phenolic acids, flavonoid glycosides, and flavonoid aglycones. Insecticidal activity was evaluated by means of contact toxicity test, which showed that all three extracts were highly effective. Pest mortality was recorded after 24 h for A. absinthium. The LC50 results revealed that the most toxic extract was that of A. absinthium (15.53 mg/cm2), followed by that of J. phoenicea (22.14 mg/cm2), and T. articulata (24.05 mg/cm2). The antifungal activities of these extracts were evaluated using the poisonous medium technique on potato dextrose agar medium. All three extracts reduced the growth of Fusarium culmorum and F. graminearum under in vitro conditions. T. articulata had the greatest inhibitory effect (55.8 %) on mycelial growth of the former. Our results reveal the potential of plant methanolic extracts to control pests common to stored grains.

The history of mass spectrometry started at the beginning of the 20th century. Since then, it has undergone an enormous expansion in terms of improvements to technical and methodological innovations, which has benefited the growth of its application in chemistry, biochemistry, pharmacology and medicine. Mass spectrometry is an analytical technique that allows molecular mass determination and structural elucidation for both known and unknown molecules. This chapter presents a general introduction to mass spectrometry instrumentation and the basic ‘know-how’ of operating principles of ionization systems and mass analysers for the study of biomolecules.

We have developed an analytical method that allows the simultaneous determination of epoxycarotenoids, hydroxycarotenoids and carotenes in monovarietal fresh homemade and industrially processed fruit products. Analyses were carried out using ultra performance liquid chromatography (UPLC). The extraction method was optimized using methanol as the first extraction solvent for lyophilized samples followed by a saponification step. Recoveries ranged between 75% and 104% depending on the compound. Repeatability was better than 10% for all compounds (%RSD, n = 3). The chromatographic analysis takes less than 17 min. In this short period, up to 27 carotenoids were identified in apple, peach and pear products. The developed method allowed us to differentiate juice from six varieties of apple by their carotenoid profile. Moreover, the methodology allows us to differentiate the carotenoid profiles from commercial juices and homemade fresh peach and pear juices, as well as to study the rearrangements of 5,6- to 5,8-epoxycarotenoids.

A fast and reliable method for the quantification of patulin using ultra high performance liquid chromatography coupled to a photodiode array detector was developed and validated for the analysis of several apple-based foodstuffs. Sample preparation was based on the QuEChERS procedure. Samples were extracted with acetonitrile and partitioned with a mixture of sodium citrate, NaCl, and Mg2SO4. The cleanup step was performed using dispersive SPE mixed with Mg2SO4 and PSA. This step was carried out twice in order to improve chromatographic results. The method was validated in spiked cloudy apple juice, apple puree, apple yogurt, beer with apple juice, and cider and applied to 24 commercial samples. The limits of detection and limits of quantification were ≥0.4 and ≥2 ng/g, respectively. Both were below the maximum legal limit established by the European Union. Recoveries for all the matrices were between 78.4 and 94.7 % while relative standard deviations were between 3.8 and 10.4 %. Patulin was detected on four apple juices from concentrate, one cloudy apple juice from an eco-store, and one beer. Nevertheless, the highest concentration found was 25.4 ng/g.