Acute inflammatory responses are protective, yet without timely resolution can lead to chronic inflammation and organ fibrosis. A systems approach to investigate self-limited (self-resolving) ...inflammatory exudates in mice and structural elucidation uncovered novel resolution phase mediators in vivo that stimulate endogenous resolution mechanisms in inflammation. Resolving inflammatory exudates and human leukocytes utilize DHA and other n−3 EFA to produce three structurally distinct families of potent di- and trihydroxy-containing products, with several stereospecific potent mediators in each family. Given their potent and stereoselective picogram actions, specific members of these new families of mediators from the DHA metabolome were named D-series resolvins (Resolvin D1 to Resolvin D6), protectins (including protectin D1–neuroprotectin D1), and maresins (MaR1 and MaR2). In this review, we focus on a) biosynthesis of protectins and maresins as anti-inflammatory–pro-resolving mediators; b) their complete stereochemical assignments and actions in vivo in disease models. Each pathway involves the biosynthesis of epoxide-containing intermediates produced from hydroperoxy-containing precursors from human leukocytes and within exudates. Also, aspirin triggers an endogenous DHA metabolome that biosynthesizes potent products in inflammatory exudates and human leukocytes, namely aspirin-triggered Neuroprotectin D1/Protectin D1 AT-(NPD1/PD1). Identification and structural elucidation of these new families of bioactive mediators in resolution has opened the possibility of diverse patho-physiologic actions in several processes including infection, inflammatory pain, tissue regeneration, neuroprotection-neurodegenerative disorders, wound healing, and others. This article is part of a Special Issue entitled “Oxygenated metabolism of PUFA: analysis and biological relevance”.
•Proresolving mediators are biosynthesized during resolution phase of acute inflammation.•Resolution is a biosynthetically active process.•Protectins and maresins are two structurally distinct families of potent local mediators.•NPD1/PD1 reduces neural inflammation, stimulates resolution and reduces pain.•MaR1 stimulates tissue regeneration and resolution mechanisms, is anti-inflammatory and reduces pain.
Polyphenols are natural phytochemicals recognized as one of the largest and most widespread classes of plant constituents occurring throughout the plant kingdom. Over the years, polyphenols have ...attracted the attention of the scientific community due to their potential health-promoting effects. Owing to their several interesting properties, polyphenols have aroused considerable interest, shedding light on their chemistry, reactivity, biological properties and their beneficial effects on both plants and humans. As a result of these investigations, a huge number of research works have been reported on polyphenols. However, despite the voluminous literature studying polyphenols and their impact on human health, there are still many issues waiting to be explored in order to explain and elucidate the positive effects of polyphenols on human health. This Special Issue was intended to gather up-to-date knowledge related to the cutting-edge research in the broad scientific area of polyphenols, providing an opportunity to show the latest advances in the exploration of polyphenols and their health beneficial effects. The special issue was a great success with more than 50 published manuscripts covering various aspects related to polyphenols and their role in human health. We are confident that this Special Issue will be of great interest to the scientific community, and will contribute to the improvement of knowledge regarding the area of polyphenols, leading to a better understanding of their role in human health.
A tryptic digest generated from Xenopus laevis fertilized embryos was fractionated by RPLC. One set of 30 fractions was analyzed by 100‐min CZE‐ESI‐MS/MS separations (50 h total instrument time), and ...a second set of 15 fractions was analyzed by 3‐h UPLC‐ESI‐MS/MS separations (45 h total instrument time). CZE‐MS/MS produced 70% as many protein IDs (4134 versus 5787) and 60% as many peptide IDs (22 535 versus 36 848) as UPLC‐MS/MS with similar instrument time (50 h versus 45 h) but with 50 times smaller total consumed sample amount (1.5 μg versus 75 μg). Surprisingly, CZE generated peaks that were 25% more intense than UPLC for peptides that were identified by both techniques, despite the 50‐fold lower loading amount; this high sensitivity reflects the efficient ionization produced by the electrokinetically pumped nanospray interface used in CZE. This report is the first comparison of CZE‐MS/MS and UPLC‐MS/MS for large‐scale eukaryotic proteomic analysis. The numbers of protein and peptide identifications produced by CZE‐ESI‐MS/MS approach those produced by UPLC‐MS/MS, but with nearly two orders of magnitude lower sample amounts.
Percolator is a widely used software tool that increases yield in shotgun proteomics experiments and assigns reliable statistical confidence measures, such as
q
values and posterior error ...probabilities, to peptides and peptide-spectrum matches (PSMs) from such experiments. Percolator’s processing speed has been sufficient for typical data sets consisting of hundreds of thousands of PSMs. With our new scalable approach, we can now also analyze millions of PSMs in a matter of minutes on a commodity computer. Furthermore, with the increasing awareness for the need for reliable statistics on the protein level, we compared several easy-to-understand protein inference methods and implemented the best-performing method—grouping proteins by their corresponding sets of theoretical peptides and then considering only the best-scoring peptide for each protein—in the Percolator package. We used Percolator 3.0 to analyze the data from a recent study of the draft human proteome containing 25 million spectra (PM:24870542). The source code and Ubuntu, Windows, MacOS, and Fedora binary packages are available from
http://percolator.ms/
under an Apache 2.0 license.
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► First comparison of UHPLC–MS/MS and capillary LC–MS for the analysis of PFCs in water. ► UHPLC–MS/MS provides the best linearity and sensitivity and shortest analysis time. ► Capillary LC–MS ...attains the best precision and comparable sensitivity. ► UHPLC–MS/MS and capillary LC–MS possess a short running time. ► Both methods were successfully applied to real samples obtaining comparable results.
In this work, the performance of ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) and capillary liquid chromatography–mass spectrometry (CLC–MS) has been studied for the analysis of eighteen perfluorinated compounds in water samples. UHPLC–MS/MS and CLC–MS analysis were carried out using a Zorbax C-18 column (50mm×2.1mm, 1.8μm) and a Zorbax SB-C18 column (150mm×0.5mm, 3.5μm), respectively, in gradient elution mode with a mobile phase of ammonium formate and methanol. Both techniques were compared with conventional LC–MS/MS in terms of speed, sensitivity, selectivity and resolution. Water samples were extracted by solid phase extraction (SPE). Mean absolute recoveries at two concentration levels, 6 and 60ngL−1, calculated from the eighteen compounds were: 77±12(s) and 82±12(s), respectively. Relative standard deviation (RSD) was lower than 16% for each perfluorinated compound. The results obtained showed that UHPLC–MS/MS and CLC–MS allow the determination of perfluorinated compounds in water samples with satisfactory sensitivity and resolution and reduced analysis time. Therefore, both techniques were employed for the analysis of water samples to determine the target analytes at low concentration levels and with lower analysis times than using HPLC–MS/MS. Similar statistical values were obtained from both techniques at 95% confidence level. Several compounds have been detected, but perfluorooctanoic acid (PFOA) (171ngL−1 and 148ngL−1 by UHPLC–MS/MS and CLC–MS, respectively) showed the highest concentration.
Olives and olive oil are two of the most important commodities produced in the Mediterranean region. Due to their significant economical importance, the usage of pesticides in their production is ...systematic, by using a wide range of plant protection products with a variety of modes of action. As a consequence, monitoring of their residue levels in this products is a necessity. In the present study a gas and liquid chromatography-tandem mass spectrometry multiresidue method, with a short sample preparation step, based on acetonitrile extraction is developed and validated according to the European Union guidelines (SANCO Doc. No. 12495/2011) in olives and olive oil, with a large scope that includes pesticides of different chemical classes. Good sensitivity and selectivity of the method were obtained with limits of quantification at 10μg/kg. All pesticides had recoveries in the range of 70–120%, with relative standard deviation values less than 20–25%, at both validation levels. Excellent linearity was achieved with r≥0,99 for both matrices. The method is easy, with low consumption of reagents, is characterized by reliability, sensitivity and therefore is suitable for the monitoring the levels of multiclass pesticides residues in olives and olive oil. The method was applied to 262 samples of the Greek market, of which 7% were found positive for the present of pesticides. In some of the samples 2–8 different analytes were detected.
•Easy and robust method using modern techniques, like GC-MSMS and LC–MSMS.•A homogenization study during sub-sampling was preformed.•Matrix effect assessment was preformed.•Validation of the method was preformed taking also into consideration EU guidelines for official controls.•Application to 262 real samples in which 7% were found positive. In some samples multiple analytes were detected.
In the field of food analysis and regulation, different instruments are used to determine the accuracy of quantification values. This is essential, as inconsistencies in values are commonly ...encountered. To visualize the degree of these discrepancies in each food matrix, we compiled a validation study based on a routine method developed in our laboratory, for 121 pesticides in six agricultural products, namely the grapefruit, potato, paprika, cabbage, spinach, and brown rice. These were analyzed by GC-MS/MS and LC-MS/MS, and the results were compared mainly on the basis of trueness. According to the results of the validation study when using GC-MS/MS, of the 121 pesticides tested in each product class, the number of analytes that satisfied the criteria of the Japanese validation guidelines was 97 in grapefruit, 111 in potato, 110 in paprika, 118 in cabbage, 111 in spinach, and 63 in brown rice. In contrast, in the analysis of the same samples by using LC-MS/MS, the number of analytes that satisfied the criteria of the validation guidelines was 50 in grapefruit, 114 in potato, 103 in paprika, 112 in cabbage, 100 in spinach, and 103 in brown rice. Inconsistences in the differences of trueness were mainly attributed to matrix effects of each instrument, as well as to food matrices, of which the most diverged matrix was that of brown rice (over 20%).
In the development of biosimilar products to Neulasta, it is essential to determine the intact molecular mass and confirm precise PEGylation sites. In this study, we applied a combination of ...techniques, including post‐column addition of triethylamine in reversed‐phase liquid chromatography–mass spectrometry (RPLC‐MS) to determine the intact molecular mass, and in‐source fragmentation (ISF) and higher‐energy collision dissociation–tandem mass spectrometry (HCD‐MS/MS) to identify the PEGylation site. Our results show that both the pegfilgrastim biosimilar candidate and Neulasta lots are mono‐PEGylated at the N‐terminal end. Furthermore, we show that the combined ISF and HCD‐MS/MS method can be used for identifying the PEGylation sites in the diPEGylated variant of pegfilgrastim. The diPEGylated variant has modification sites at the N‐terminal end and a lysine at position 35 in the protein sequence.
•Analysis method for 91 GC-, 81 LC amenable pesticides and 6 indicator PCBs in eggs.•Sample preparation: MSPD, small-scale GPC, SPE for GC and LC analytes, MS/MS.•Confirmation of the clean-up effect ...due to TLC for different lipid classes.•164 of 178 analytes fulfilled the validation criteria in hen eggs.•Successful investigation of EU proficiency test material (egg, poultry meat, cream).
A sample preparation method was developed for the analysis of chicken eggs to determine 97 GC and 81 LC amenable residues, including organophosphates, organochlorines, pyrethroids, triazoles, carboxyl-containing compounds, and the indicator PCBs. Hereby, considerations were given to the recoveries of the analytes, the method's suitability for routine analysis, and the assessment of the clean-up effect, for which a simple thin layer chromatography was implemented to visualize the most important lipid classes.
The procedure consisted of (I) the extraction by matrix solid phase dispersion, and the clean-up by means of (II) small-scale gel permeation chromatography (GPC) and (III) two different solid phase extractions (SPE) for GC and LC amenable analytes, as well as (IV) the quantification using GC–MS/MS and LC–MS/MS. Cyclohexane/ethyl acetate was chosen as extraction solvent due to its suitability for extracting strong non-polar but also more polar analytes. The classical GPC was scaled down to ensure a 50% lower solvent consumption. The comprehensive investigation of conventional and modern zirconium-oxide-coated SPE materials resulted in the selection of octadecyl-modified silica (C18) combined with primary secondary amine using acetonitrile as elution solvent for GC amenable analytes and pure C18 in combination with acidified methanol for LC amenable pesticides.
Compared to the currently established EN 1528 method the sample preparation proposed offered a higher sample throughput and a lower solvent consumption. Furthermore, for the first time the clean-up effectiveness of the sample preparation steps was documented as shown by means of thin-layer chromatography.
The validation of chicken eggs proved the fulfillment of the quality control criteria for 164 of the 178 analytes tested, mostly at the lowest validated level of 5μg/kg for pesticides and 0.5μg/kg for the single indicator PCBs. However, the analysis of strongly polar analytes was still problematic, which could be attributed to the extraction and the GPC step. Nevertheless, the successful investigation of EU proficiency test materials (EUPT AO 07–09) confirmed the comparability of the results with the currently established sample preparation procedures and demonstrated the potential of the applicability of the presented method to other matrices as exemplified for lean poultry meat and fatty liquid cream.
This work summarized all results obtained during almost two-years of a monitoring programme carried out in five municipal sewage treatment plants (STPs) located in the north, centre and south-east of ...Spain. The study evaluated the occurrence and persistence of a group of 100 organic compounds belonging to several chemical groups (pharmaceuticals, personal care products, pesticides and metabolites). The average removal efficiencies of the STPs studied varied from 20% (erythromycin) to 99% (acetaminophen). In analysed samples, we identified a large number of compounds at mean range concentrations between 7–59,495 ng/L and 5–32,720 ng/L for influent and effluent samples, respectively. This study also identified 20 of the mostly detected and persistent compounds in wastewater effluent, of which hydrochlorothiazide, atenolol, gemfibrozil, galaxolide and three metabolites (fenofibric acid, 4-AAA and 4-FAA), presented the highest average contribution percentages, in relation to the total load of contaminants for the different STPs effluent studied.
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► The results summarize two-years of a monitoring programme. ► 100 organic compounds (priority substances and emerging contaminants) were analysed. ► The removal efficiency of 5 STPs of Spain was evaluated. ► The presence of target compounds in treated wastewater was also checked. ► The most frequently drugs detected were: antibiotics<anti-inflammatories<β-blockers.
Antibiotics and analgesics/anti-inflammatories were the most frequently drugs detected, following by some β-blockers, synthetic fragrances, lipid regulators and diuretics.
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