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dr hab. inż. Weronika Hewelt-Belka
Employment
- Associate professor at Department of Analytical Chemistry
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total: 38
Catalog Publications
Year 2024
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Keratinocyte-derived small extracellular vesicles supply antigens for CD1a-resticted T cells and promote their type 2 bias in the context of filaggrin insufficiency
PublicationIntroduction: Exosome-enriched small extracellular vesicles (sEVs) are nanosized organelles known to participate in long distance communication between cells, including in the skin. Atopic dermatitis (AD) is a chronic inflammatory skin disease for which filaggrin (FLG) gene mutations are the strongest genetic risk factor. Filaggrin insufficiency affects multiple cellular function, but it is unclear if sEV-mediated cellular communication...
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Microbe Cultivation Guidelines to Optimize Rhamnolipid Applications
PublicationIn the growing landscape of interest in natural surfactants, selecting the appropriate one for specific applications remains challenging. The extensive, yet often unsystematized, knowledge of microbial surfactants, predominantly represented by rhamnolipids (RLs), typically does not translate beyond the conditions presented in scientific publications. This limitation stems from the numerous variables and their interdependencies...
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Modification of gradient HPLC method for determination of small molecules' affinity to human serum albumin under column safety conditions: Robustness and chemometrics study
PublicationIn the early stages of drug discovery, beyond the biological activity screening, determining the physicochemical properties that affect the distribution of molecules in the human body is an essential step. Plasma protein binding (PPB) is one of the most important investigated endpoints. Nevertheless, the methodology for measuring %PPB is significantly less popular and standardized than other physicochemical properties, like lipophilicity....
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Novel materials and approaches for solid-phase (micro) extraction in LC-MS-based metabolomics
PublicationMetabolomics offers a unique approach to study biological systems. Liquid chromatography coupled to mass spectrometry serves as a powerful analytical tool to analyze complex mixtures of metabolites. However, successful metabolomic analysis relies on sample preparation. While untargeted analysis traditionally favors nonselective procedures like liquid-liquid extraction (LLE), these methods can compromise analytical performance due...
Year 2023
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A comprehensive lipidomic analysis of oilseeds using LC-Q-TOF-MS and dispersive micro-solid phase (D-μ-SPE) extraction techniques
PublicationGrowing public health awareness increased the demand for natural products with positive health benefits. Vegetable oils containing omega-3 fatty acids (FA) such as alpha-linolenic acid (ALA) has been proposed as a possible source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). In this work, a comprehensive lipidome analysis and comparison of seven oilseeds, including flaxseed, chia, hemp, black cumin, white sesame,...
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Characterization of GM3 Gangliosides in Human Milk throughout Lactation: Insights from the Analysis with the Use of Reversed-Phase Liquid Chromatography Coupled to Quadrupole Time-Of-Flight Mass Spectrometry
PublicationGangliosides are complex lipids found in human milk that play important structural and biological functions. In this study, we utilized reversed-phase liquid chromatography coupled to quadrupole time-of-flight mass spectrometry to evaluate the molecular distribution of GM3 in human milk samples collected at distinct lactation stages, ranging from colostrum to advanced lactation samples. Throughout lactation, GM3 d40:1 emerged as...
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Direct injection liquid chromatography-mass spectrometry (DI-LC-MS) analysis for rapid lipidomic profiling of extracellular vesicles
PublicationExtracellular vesicles (EVs) are small, spherical particles produced by eukaryotic and prokaryotic cells, surrounded by a bilayer membrane and carrying various bioactive molecules, such as proteins, surface receptors, membrane and soluble proteins, lipids, and nucleic acids. EVs are of substantial interest because of their important roles in cell communication, epigenetic regulation and possible application in disease diagnosis...
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O07 Filaggrin insufficiency renders keratinocyte-derived small extracellular vesicles capable of affecting CD1a-mediated T-cell responses and promoting allergic inflammation
PublicationThe association between FLG loss-of-function mutations and the development of atopic dermatitis (AD) indicates that filaggrin is critical to skin barrier function; the mutations are also linked to additional allergic manifestations, but it is unknown how the skin may influence inflammation in distant tissues. Here we investigated the impact of filaggrin insufficiency on keratinocyte-derived exosome-enriched small extracellular...
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Pre-analytical aspects in metabolomics of human biofluids – sample collection, handling, transport, and storage
PublicationMetabolomics is the field of omics research that offers valuable insights into the complex composition of biological samples. It has found wide application in clinical diagnostics, disease investigation, therapy prediction, monitoring of treatment efficiency, drug discovery, or in-depth analysis of sample composition. A suitable study design constitutes the fundamental requirements to ensure robust and reliable results from the...
Year 2022
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A method for a comprehensive lipidomic analysis of flaxseed (Linum usitatissimum) with the use of LC–Q-TOF-MS and dispersive micro-solid-phase (μDSPE) extraction
PublicationFlaxseed (FS) is one of the richest sources of α-linolenic acid oil and lignans, and it is suggested that the consumption of flaxseed may contribute to the prevention of certain chronic diseases such as many types of cancer, diabetes, cardiovascular diseases and cerebrovascular stroke. Here, we demonstrate a new method for comprehensive FS lipidome profiling with the use of LC–Q-TOF-MS and dispersive micro-solid-phase extraction....
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IFI16 Impacts Metabolic Reprogramming during Human Cytomegalovirus Infection
PublicationCellular lipid metabolism plays a pivotal role in human cytomegalovirus (HCMV) infection, as increased lipogenesis in HCMV-infected cells favors the envelopment of newly synthesized viral particles. As all cells are equipped with restriction factors (RFs) able to exert a protective effect against invading pathogens, we asked whether a similar defense mechanism would also be in place to preserve the metabolic compartment from HCMV...
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The Influence of Storage on Human Milk Lipidome Stability for Lipidomic Studies
PublicationHuman milk (HM) lipidome stability during storage is crucial in lipidomic studies to avoid misinterpretations. Facing the lack of comprehensive work on the HM lipidome stability, we performed a study on a potential alteration in the lipid profiles of HM samples stored under different conditions. An untargeted LC-Q-TOF-MS-based approach was applied to study the influence of storage conditions as well as the interaction of the storage...
Year 2021
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Analytical Strategies and Applications in Lipidomics
PublicationLipidomics is a field of study aiming at a comprehensive analysis of a set of lipids in biological systems. This includes both quantitative and qualitative characterization of lipid profiles, comparative studies of lipidomes, and analysis of lipid-protein interactions. Lipids are structurally diversified that make their analysis a challenging task. The constant advances in analytical technologies, especially in the field of high-performance...
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Sample Preparation in Foodomics: Miniaturized Solid-Phase Extraction
PublicationAnalytical chemists face a challenge to bring comprehensive information on a given food and biological sample by using the best available analytical techniques and meet the requirements of sustainable development and green chemistry at the same time. A key objective of this chapter is to review selected literature data on the utilization of solid-phase extraction techniques with special attention to their miniaturized modes in...
Year 2020
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Comparative Lipidomic Study of Human Milk from Different Lactation Stages and Milk Formulas
PublicationIn this report, we present a detailed comparison of the lipid composition of human milk (HM) and formula milk (FM) targeting different lactation stages and infant age range. We studied HM samples collected from 26 Polish mothers from colostrum to 19 months of lactation, along with FM from seven brands available on the Polish market (infant formula, follow-on formula and growing-up formula). Lipid extracts were analysed using liquid...
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Lipidomika – strategie analityczne i zastosowania
PublicationBioanalityka, to interdyscyplinarna dziedzina wiedzy, która stanowi szybko rozwijający się obecnie dział chemii analitycznej. Bioanaliza zaczyna odgrywać kluczową rolę w szybko rozwijających się dziedzinach współczesnej bionauki w ramach genomiki, proteomiki, metabolomiki, lipidomiki i innych. Umiejętność doboru odpowiednich metod i narzędzi, w zależności od rodzaju podejmowanego problemu, jest niezwykle ważne i często decyduje...
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Nuclear Magnetic Resonance Metabolomics Reveals Qualitative and Quantitative Differences in the Composition of Human Breast Milk and Milk Formulas
PublicationCommercial formula milk (FM) constitutes the best alternative to fulfill the nutritional requirements of infants when breastfeeding is precluded. Here, we present the comparative study of polar metabolite composition of human breast milk (HBM) and seven different brands of FM by nuclear magnetic resonance spectroscopy. The results of the multivariate data analysis exposed qualitative and quantitative differences between HBM and...
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The dispersive micro-solid phase extraction method for MS-based lipidomics of human breast milk*
PublicationA simple and rapid microextraction method ensuring high lipidome coverage was developed for liquid chromatography mass spectrometry (LC-MS)-based lipidomics of human breast milk. The dispersive microsolid phase extraction (D-µ-SPE) technique, coupled with the design of experiment (DoE) method, enabled the study of the influence of several conditions (desorption solvent, stationary phase ratio, and sorption and desorption time)...
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Untargeted Lipidomics Analysis of the Cyanobacterium Synechocystis sp. PCC 6803: Lipid Composition Variation in Response to Alternative Cultivation Setups and to Gene Deletion
PublicationCyanobacteria play an important role in several ecological environments, and they are widely accepted to be the ancestors of chloroplasts in modern plants and green algae. Cyanobacteria have become attractive models for metabolic engineering, with the goal of exploring them as microbial cell factories. However, the study of cyanobacterial lipids’ composition and variation, and the assessment of the lipids’ functional and structural...
Year 2019
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A new dilution-enrichment sample preparation strategy for expanded metabolome monitoring of human breast milk that overcomes the simultaneous presence of low- and high-abundance lipid species
PublicationThe complex nature of human breast milk (HBM) makes samples difficult to analyze, requiring several extraction techniques and analytical platforms to obtain high metabolome coverage. In this work, we combined liquid-liquid extraction (LLE) and solid-phase extraction (SPE) techniques to prepare HBM samples to overcome the challenge of low- and high-abundance lipid species, enabling the semiquantitative analysis of total HBM lipids...
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State of the art in sample preparation for human breast milk metabolomics—merits and limitations
PublicationHuman breast milk (HBM) contains metabolites that are diverse in chemical structure and concentration. These metabolites range from high-abundance to low-abundance compounds and from polar to nonpolar. This complex nature of HBM makes HBM samples difficult to analyze, especially in global metabolomic studies for the determination and characterization of complete sets of metabolites. In this review, the state of the art in sample...
Year 2018
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Chemistry of human breast milk – a comprehensive review of the composition and role of milk metabolites in child development
PublicationEarly nutrition has an enormous influence on a child's physiological function, immune system maturation and cognitive development. Human breast milk (HBM) is recognized as the gold standard for human infant nutrition. Accordingly to WHO report breastfeeding is consider as an unequalled way of providing ideal food to the infant, which is required for his healthy growth and development. HBM contains various macronutrients (carbohydrates,...
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New sample preparation strategies for comprehensive lipidomics of human breast milk
PublicationGLOBAL PROFILING ANALYSIS OF HUMAN BREAST MILK (HBM) LIPIDS IS HINDERED BY COMPLEX COMPOSITION OF HBM. THIS PROBLEM IS HUGE FROM ANALYTICAL POINT OF VIEW, SINCE DUE TO THAT DETECTION OF ALL HBM LIPIDS DURING ONE ANALYTICAL RUN IS LIMITED. THUS, SAMPLE PREPARATION STEP CONSTITUTES A CRUCIAL STEP IN UNTARGETED LIPIDOMIC ANALYSIS OF HBM, ESPECIALLY WHEN SEMI-QUANTITATIVE ANALYSIS IS ASSUMED. HEREIN, WE PRESENT A COMPARISON OF PUBLISHED...
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Strategia analizy niecelowanej w metabolomice i lipidomice – aspekty praktyczne cz.2
PublicationStrategia analizy niecelowanej jest powszechnie wykorzystywana do analizy różnego rodzaju próbek biologicznych, takich jak tkanki czy płyny ustrojowe, w celu otrzymania „odcisku palca” oraz określenia różnic w składzie jakościowym oraz ilościowym pomiędzy badanymi próbkami. Wskazanie różnic w zawartości różnych biocząsteczek (peptydów, metabolitów, lipidów) pozwala na uzyskanie informacji na temat powiązania składu biochemicznego...
Year 2017
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Lipidomika mleka kobiecego - wiele do odkrycia.
PublicationMleko kobiece jest wieloskładnikową wydzieliną, której skład zależy od wielu czynników, dlatego różni się międzyosobniczo, jest niepowtarzalne i oryginalne. W każdym mleku można wyróżnić zarówno składniki odżywcze niezbędne do prawidłowego wzrostu niemowlęcia, jak i bioaktywne substancje chroniące niemowlę przed zakażeniami, stanami zapalnymi oraz przyczyniającymi się do dojrzewania układu odpornościowego Ze względu na zawartość...
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Rapid Characterization of the Human Breast Milk Lipidome Using a Solid-Phase Microextraction and Liquid Chromatography-Mass Spectrometry-Based Approach.
PublicationHuman breast milk (HBM) is a biofluid consisting of various biomolecules such as proteins, lipids, carbohydrates, minerals and bioactive substances. Due to its unique and complex composition, HBM provides not only nutritional components required for the growth of the infant, but also additional protection against infections. Global insight into the composition of HBM is crucial to understanding the health benefits infants receive...
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Strategia analizy niecelowanej w metabolomice i lipidomice. Cz. 1. Aspekty praktyczne
PublicationStrategia analizy niecelowanej jest powszechnie wykorzystywana do analizy różnego rodzaju próbek biologicznych, takich jak tkanki czy płyny ustrojowe, w celu otrzymania „odcisku palca” oraz określenia różnic w składzie jakościowym oraz ilościowym pomiędzy badanymi próbkami. Wskazanie różnic w zawartości różnych biocząsteczek (peptydy, metabolity, lipidy) pozwala na uzyskanie informacji na temat powiązania składu biochemicznego...
Year 2016
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Development of chromatographic and mass spectrometry conditions for untargeted lipidomic analysis of human breast milk
PublicationHuman breast milk (HBM) is a complex biofluid containing components which are essential for the proper growth of the newborn (for example nutrients such as carbohydrates, proteins, lipids) and also bioactive substances. According to the literature, most of studies involving HBM nutrients investigations focused essentially on the determination of proteins. The molecular composition of lipids in HBM is not well known as the protein...
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Forced degradation studies of ivabradine and in silico toxicology predictions for its new designated impurities
PublicationAll activities should aim to eliminate genotoxic impurities and/or protect the API againstdegradation. There is a necessity to monitor impurities from all classification groups,hence ivabradine forced degradation studies were performed. Ivabradine was provedto be quite durable active substance, but still new and with insufficient stability data.Increased temperature, acid, base, oxidation reagents and...
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Forced Degradation Studies of Ivabradine and In Silico Toxicology Predictions for Its New Designated Impurities
PublicationAll activities should aim to eliminate genotoxic impurities and/or protect the API against degradation. There is a necessity to monitor impurities from all classification groups, hence ivabradine forced degradation studies were performed. Ivabradine was proved to be quite durable active substance, but still new and with insufficient stability data. Increased temperature, acid, base, oxidation reagents and light were found to cause...
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Lipidomics of Staphylococcus aureus – a new insight into the antibiotic resistant phenotype
PublicationStaphylococcus aureus is an aggressive pathogen responsible for a variety of diseases, in cluding life-threatening sepsis. Many strains of these bacteria are resistant to multiple classes of antibiotics, which is a substantial clinical problem in the treatment of S. aureus infections. We developed a comprehensive untargeted lipidomic workflow, including sample preparation, liquid chromatography-quadrupole-time-of-flight mass spectrometry...
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Photoinactivation of Staphylococcus aureus using protoporphyrin IX: the role of haem-regulated transporter HrtA
PublicationLight- and photosensitiser-based antimicrobial photodynamic therapy is a very promising approach to the control of microbial infections. How the phenotypic features of a microorganism affect its response to photosensitiser-based photokilling represents an area of substantial research interest. To understand the mechanisms governing the phenomenon of a strain-dependent response to photodynamic inactivation (PDI), we analysed the...
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Untargeted Lipidomics Reveals Differences in the Lipid Pattern among Clinical Isolates of Staphylococcus aureus Resistant and Sensitive to Antibiotics
PublicationStaphylococcus aureus resistance to antibiotics is a significant clinical problem worldwide. In this study, an untargeted lipidomics approach was used to compare the lipid fingerprints of S. aureus clinical isolates that are resistant and sensitive to antibiotics. High-performance liquid chromatography coupled with time-of-flight mass spectrometry was employed to rapidly and comprehensively analyze bacterial lipids. Chemometric...
Year 2015
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Prediction of Overall In Vitro Microsomal Stability of Drug Candidates Based on Molecular Modeling and Support Vector Machines. Case Study of Novel Arylpiperazines Derivatives
PublicationOther than efficacy of interaction with the molecular target, metabolic stability is the primary factor responsible for the failure or success of a compound in the drug development pipeline. The ideal drug candidate should be stable enough to reach its therapeutic site of action. Despite many recent excellent achievements in the field of computational methods supporting drug metabolism studies, a well-recognized procedure to model...
Year 2014
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Chemia analityczna w lipidomice
PublicationLipidomika, stanowiąca gałąź metabolomiki, jest prężnie rozwijającą się dziedziną nauki, w której badania ukierunkowane są na identyfikację oraz oznaczenie ilościowe lipidów komórkowych, poznanie ich funkcji biologicznych oraz oddziaływań z innymi cząsteczkami. Wzrost zainteresowania badaniami lipidomicznymi w ostatnich latach, zwłaszcza w obszarze tworzenia profili lipidów komórkowych, związany był z wprowadzeniem do praktyki...
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Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry
PublicationStaphylococcus aureus is a common pathogen known to cause relatively minor infections as well as severe disorders in humans. Although there is fair amount of published data concerning various aspects of its biology, epidemiology, genetics, etc., there is still a scarce amount of data presenting reliable and thorough investigations regarding high-throughput analysis of total S. aureus lipid content. Therefore, the aim of this study...
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Electrochemical simulation of enzymatic transformations studied for the selected antitumor acridine derivatives
PublicationThe elucidation of the metabolic pathways and the biotransformation mechanisms of potential drugs is a crucial point in drug development. It allows to know the activation routes of the new biologically active compounds, especially in respect to their possible toxicity. Generally, in vivo or in vitro experiments with liver microsomes or hepatocytes are performed. However, these testing schemes are tedious, time consuming and of...
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Mass spectrometry based identification of geometric isomers during metabolic stability study of a new cytotoxic sulfonamide derivatives supported by quantitative structure-retention relationships
PublicationA set of 15 new sulphonamide derivatives, presenting antitumor activity have been subjected to a metabolic stability study. The results showed that besides products of biotransformation, some additional peaks occurred in chromatograms. Tandem mass spectrometry revealed the same mass and fragmentation pathway, suggesting that geometric isomerization occurred. Thus, to support this hypothesis, quantitative structure-retention relationships...
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