Comprehensive identification of native medium-sized and short bioactive peptides in sea bass muscle

Native peptides from sea bass muscle were analyzed by two different approaches: medium-sized peptides by peptidomics analysis, whereas short peptides by suspect screening analysis employing an inclusion list of exact m/z values of all possible amino acid combinations (from 2 up to 4). The method was also extended to common post-translational modifications potentially interesting in food analysis, as well as non-proteolytic aminoacyl derivatives, which are well-known taste-active building blocks in pseudo-peptides. The medium-sized peptides were identified by de novo and combination of de novo and spectra matching to a protein sequence database, with up to 4077 peptides (2725 modified) identified by database search and 2665 peptides (223 modified) identified by de novo only; 102 short peptide sequences were identified (with 12 modified ones), and most of them had multiple reported bioactivities. The method can be extended to any peptide mixture, either endogenous or by protein hydrolysis, from other food matrices. © 2020 Elsevier Lt

Degradation of the polar lipid and fatty acid molecular species in extra virgin olive oil during storage based on shotgun lipidomics

Among the bioactive compounds present in extra-virgin olive oil, polar lipids and free fatty acids are minor compounds with well-known nutritional values and have been studied for traceability and adulteration investigations as well. In the present paper, the simultaneous characterization of polar lipids and free fatty acids in a pool of fifteen EVOO samples was achieved by means of reversed phase C18 analysis coupled to negative polarity high-resolution mass spectrometry. A total of 24 polar lipids, comprising 19 phospholipids and 5 sulfolipids, and 27 free fatty acids were tentatively identified, including several odd-chain and very long-chain fatty acids at trace levels. Moreover, a one-month study of lipid degradation on simulated storage conditions was carried out thanks to the set-up of a dedicated approach for degradation product analysis which was implemented of Compound Discoverer software. By virtue of the customized data processing workflow, more than forty compounds were tentatively identified, including compounds deriving from hydrolysis and oxidation reactions. Finally, by analysis of peak area trends, phosphoester hydrolyses of polar heads of phospholipids emerged as the fastest reactions, followed by glycerol ester hydrolyses and oxidative processes

Investigating the Short Peptidome Profile of Italian Dry-Cured Ham at Different Processing Times by High-Resolution Mass Spectrometry and Chemometrics

Short peptides have been spiking interest owing to their significant contribution to the taste and functional properties of dry-cured ham. In this study, a suspect screening approach based on high-resolution mass spectrometry was employed for the comprehensive characterization of the short endogenous peptidome in dry-cured ham samples at different processing stages (14, 22, and 34 months). After careful manual spectra interpretation, a chemometric approach based on principal component analysis was employed for highlighting the differences between the three sets of samples. A total of 236 short peptide sequences was tentatively identified, including 173 natural short peptides and 63 sequences containing non-proteinogenic amino acids, the highest number ever reported for endogenous sequences in dry-cured ham. Samples in the latest processing stages presented a generally higher abundance of dipeptides, indicating residual proteolytic activity. Moreover, the several annotated modified short peptides, mainly pyroglutamination and lactoyl conjugation, allowed hypothesizing several reactions occurring over time. For the first time, several lactoyl-dipeptides were tentatively identified in dry-cured ham samples with maximum concentration in the late processing stage samples. The presented results significantly contribute to the understanding of the reaction involving short peptides that affect the sensory and functional properties of dry-cured ham

Profiling and quantitative analysis of underivatized fatty acids in Chlorella vulgaris

Chlorella vulgaris is a popular microalga used for biofuel production; nevertheless, it possesses a strong cell wall that hinders the extraction of molecules, especially lipids within the cell wall. For tackling this issue, we developed an efficient and cost‐effective method for optimal lipid extraction. Microlaga cell disruption by acid hydrolysis was investigated comparing different temperatures and reaction times; after hydrolysis, lipids were extracted with n‐hexane. The best recoveries were obtained at 140°C for 90 min. The microalgae were then analyzed by an untargeted approach based on liquid chromatography with high‐resolution mass spectrometry, providing the tentative identification of 28 fatty acids. First, a relative quantification on the untargeted data was performed using peak area as a surrogate of analyte abundance. Then, a targeted quantitative method was validated for the tentatively identified fatty acids, in terms of recovery (78‐100%), intra‐ and interday relative standard deviations ( 0.98). The most abundant fatty acids were palmitic, palmitoleic, oleic, linoleic, linolenic, and stearic acids

Andean blueberry of the Genus Disterigma: a high-resolution mass spectrometric approach for the comprehensive characterization of phenolic compounds

Wild neotropical blueberries, endemic of Central and South American areas, are promising yet still undisclosed sources of bioactive compounds. Most research studies have addressed wild and cultivated blueberries from Europe and North America, despite the extremely wide variety of wild neotropical species. In the present paper, for the first time, the phenolic composition of Disterigma alaternoides was investigated through ultra-high-performance liquid chromatography coupled to high-resolution mass-spectrometric analysis followed by accurate data analysis and compound validation with a dedicated structure-based workflow. D. alaternoides, which belongs to a closely related genus to that of the common blueberry, grows exclusively in the Andean regions over 2000 above sea level. Thanks to the dedicated analytical platform, 249 phenolic compounds were tentatively identified, including several anthocyanins, flavonoids, phenolic acids, and proanthocyanidins. The nature and heterogeneity of identified phenolic compounds demonstrate once more the need for a more profound knowledge of such still uncharted matrice

Detailed investigation of the composition and transformations of phenolic compounds in fresh and fermented Vaccinium floribundum berry extracts by high-resolution mass spectrometry and bioinformatics

Introduction:Blueberries are known for their very high content of biologically activephenolic compounds; nonetheless, differently from the North American andEuropean species of blueberries, Neotropical blueberries have not been extensivelystudied yet.Objectives:In the present paper, the phenolic composition ofVacciniumfloribundumKunth, which is endemic to the Andean regions and grows 1,600to 4,500 meters above sea level, was investigated by ultra-high-performanceliquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). Native and fermented berries were compared in terms of phenoliccomposition as well as antioxidant activity, total phenolic content, and totalanthocyanin content.Materials and Methods:V. floribundumnative and fermented berries were extractedand analyzed by UHPLC-HRMS. The acquired datasets were processed by Com-pound Discoverer 3.1 using a dedicated data analysis workflow that was specificallyset up for phenolic compound identification.Results:In total, 309 compounds were tentatively identified, includinganthocyanins, flavonoids, phenolic acids, and proanthocyanidins. The moleculartransformations of phenolic compounds during fermentation were comprehen-sively investigated for the first time, and by a customized data processingworkflow, 13 quinones and quinone methides were tentatively identified in thefermented samples. Compared to other species of the genusVaccinium, a peculiarphenolic profile is observed, with low abundance of highly methylatedcompounds

Production and Characterization of Medium-Sized and Short Antioxidant Peptides from Soy Flour-Simulated Gastrointestinal Hydrolysate

Soybeans (Glycine max) are an excellent source of dietary proteins and peptides with potential biological activities, such as antihypertensive, anti-cholesterol, and antioxidant activity; moreover, they could prevent cancer. Also, soy contains all the essential amino acids for nutrition; therefore, it represents an alternative to animal proteins. The goal of this paper was the comprehensive characterization of medium-sized and short peptides (two to four amino acids) obtained from simulated gastrointestinal digestion. Two different analytical approaches were employed for peptide characterization, namely a common peptidomic analysis for medium-sized peptides and a suspect screening analysis for short peptides, employing an inclusion list of exact m/z values of all possible amino acid combinations. Moreover, fractionation by preparative reversed-phase liquid chromatography was employed to simplify the starting protein hydrolysate. Six fractions were collected and tested for antioxidative activity by an innovative antioxidant assay on human gastric adenocarcinoma AGS cell lines. The two most active fractions (2 and 3) were then characterized by a peptidomic approach and database search, as well as by a suspect screening approach, in order to identify potential antioxidant amino acid sequences. Some of the peptides identified in these two fractions have been already reported in the literature for their antioxidant activity

Untargeted analysis of environmental contaminants in surface snow samples of Svalbard Islands by liquid chromatography-high resolution mass spectrometry

In recent years, there is increasing attention on the contaminants of emerging concern (CECs), which include plasti-cizers, flame retardants, industrial chemicals, pharmaceuticals, and personal care products, since they have been de-tected even far away from pollution sources. The polar regions are not exempt from the presence of anthropogenic contaminants, and they are employed as a model for understanding the pollutant fate and impact.During the 2021 spring campaign, sixteen surface snow samples were collected close to the research station of Ny-Alesund located on the Spitsbergen Island of the Norwegian Svalbard Archipelago. The samples were extracted by solid-phase extraction and analyzed by liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS) following an untargeted approach. Compound tentative identification was obtained with the aid of the software Compound Discoverer, using both mass spectral database search and manual validation. Among the 114 com-pounds identified with a high confidence level in the snow samples, >80 have some commercial or industrial use (drugs, plasticizers, fragrances, etc.), therefore they could be of anthropogenic origin. Nonetheless, a clear contamina-tion trend did not appear in the snow samples collected on eight different days during one month. The comparison with aerosol samples collected in the same area did not help identifying the source, either, since only a few compounds were in common, and they were mainly of natural origin. As such, the analysis of aerosol sample did not support possible long-range transport, also considering that compounds were detected mostly in the coarse fraction