Journal of separation science最新文献

{"title":"Chiral Separation and Determination of Multiple Organophosphorus Pesticide Enantiomers in Soil Based on Cellulose-Based Chiral Column by LC–MS/MS","authors":"Liang Li,&nbsp;Rendan Zhou,&nbsp;Huiying Xie,&nbsp;Gang Li,&nbsp;Zhiguang Xu,&nbsp;Meijia Liu,&nbsp;Wanting Gao","doi":"10.1002/jssc.70100","DOIUrl":"https://doi.org/10.1002/jssc.70100","url":null,"abstract":"<div>\u0000 \u0000 <p>The widespread use of organophosphorus pesticides (OPs) has raised significant environmental and health concerns due to their residues in soil and potential entry into the food chain. This study introduced chiral analysis methods for four OPs—methamidophos (METHP), dipterex (DIP), malathion (MALA), and isothiophos-methyl (ISOME)—using liquid chromatography–tandem mass spectrometry (LC–MS/MS) with cellulose-based chiral columns. Three distinct methods were established: one for METHP, another for DIP, and a third for MALA and ISOME. Key chromatographic variables, including organic mobile phases and column temperatures, were systematically optimized, achieving maximum resolutions (<i>R</i>s) of 1.61 for METHP, 2.40 for DIP, 1.70 for MALA, and 2.02 for ISOME. The QuEChERS method was employed for sample pretreatment, ensuring high recoveries. All three methods demonstrated excellent linearity (<i>R</i> &gt; 0.998), accuracy with recoveries ranging from 79% to 121%, precision with RSD% &lt; 11%, and sensitivity with low limits of enantiomer detection (LODs) as low as 0.17 µg/kg for METHP, 0.087 µg/kg for DIP, 0.062 µg/kg for MALA, and 0.054 µg/kg for ISOME, representing a sensitivity improvement of 16–172 times compared to existing methods. Field soil samples from Yangzizhou District, Nanchang, China, revealed significant contamination by ISOME, with concentrations of a single enantiomer reaching up to 8343 µg/kg, while MALA exhibited varying enantiomeric ratios with depth. This study provides robust analytical tools for monitoring chiral OP residues in soil, contributing to food safety and environmental protection.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143438959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From Complexity to Clarity: Expanding Metabolome Coverage With Innovative Analytical Strategies","authors":"Kanukolanu Aarika,&nbsp;Ramijinni Rajyalakshmi,&nbsp;Lakshmi Vineela Nalla,&nbsp;Siva Nageswara Rao Gajula","doi":"10.1002/jssc.70099","DOIUrl":"https://doi.org/10.1002/jssc.70099","url":null,"abstract":"<p>Metabolomics, a powerful discipline within systems biology, aims at comprehensive profiling of small molecules in biological samples. The challenges of biological sample complexity are addressed through innovative sample preparation methods, including solid-phase extraction and microextraction techniques, enhancing the detection and quantification of low-abundance metabolites. Advances in chromatographic separation, particularly liquid chromatography (LC) and gas chromatography (GC), coupled with high-resolution (HR) mass spectrometry (MS), have significantly improved the sensitivity, selectivity, and throughput of metabolomic studies. Cutting-edge techniques, such as ion-mobility mass spectrometry (IM-MS) and tandem MS (MS/MS), further expand the capacity for comprehensive metabolite profiling. These advanced analytical platforms each offer unique advantages for metabolomics, with continued technological improvements driving deeper insights into metabolic pathways and biomarker discovery. By providing a detailed overview of current trends and techniques, this review aims to offer valuable insights into the future of metabolomics in human health research and its translational potential in clinical settings. Toward the end, this review also highlights the biomedical applications of metabolomics, emphasizing its role in biomarker discovery, disease diagnostics, personalized medicine, and drug development.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70099","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography","authors":"Sovannychloé Nai,&nbsp;Christiane Adrielly Alves Ferraz,&nbsp;Thomas Gaslonde,&nbsp;Jackson Roberto Guedes da Silva Almeida,&nbsp;Edilson Beserra de Alencar Filho,&nbsp;Cíntia Emi Yanaguibashi Leal,&nbsp;Benjamin Musnier,&nbsp;Laurent Picot,&nbsp;Pierre-Edouard Bodet,&nbsp;Marie-Christine Lallemand,&nbsp;Raphaël Grougnet,&nbsp;Raimundo Gonçalves de Oliveira Junior","doi":"10.1002/jssc.70094","DOIUrl":"https://doi.org/10.1002/jssc.70094","url":null,"abstract":"<p>Favelines, benzocycloheptene diterpenes exclusively found in <i>Cnidoscolus quercifolius</i>, have shown promising anticancer properties, particularly phyllacanthone. Nevertheless, the isolation of these compounds remains challenging due to their similar polarity. The use of centrifugal partition chromatography (CPC) has enabled the quick and efficient purification of phyllacanthone from <i>C. quercifolius</i> crude extract with a yield of 4.5% (w/w) and a purity of 83%. Favelines-enriched fractions were also obtained and purified by preparative HPLC, resulting in the isolation of faveline, isofavelol, and deoxofaveline. In addition, phyllacanthone B, a previously unreported derivative, was identified and characterized. Pharmacological evaluations revealed the significant anti-melanoma activity of purified metabolites, with distinct structure–activity relationships. These results shed light on the mechanism of action of favelines, suggesting their potential as hits for the development of novel anticancer agents.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143404321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography","authors":"Sovannychloé Nai,&nbsp;Christiane Adrielly Alves Ferraz,&nbsp;Thomas Gaslonde,&nbsp;Jackson Roberto Guedes da Silva Almeida,&nbsp;Edilson Beserra de Alencar Filho,&nbsp;Cíntia Emi Yanaguibashi Leal,&nbsp;Benjamin Musnier,&nbsp;Laurent Picot,&nbsp;Pierre-Edouard Bodet,&nbsp;Marie-Christine Lallemand,&nbsp;Raphaël Grougnet,&nbsp;Raimundo Gonçalves de Oliveira Junior","doi":"10.1002/jssc.70094","DOIUrl":"https://doi.org/10.1002/jssc.70094","url":null,"abstract":"<p>Favelines, benzocycloheptene diterpenes exclusively found in <i>Cnidoscolus quercifolius</i>, have shown promising anticancer properties, particularly phyllacanthone. Nevertheless, the isolation of these compounds remains challenging due to their similar polarity. The use of centrifugal partition chromatography (CPC) has enabled the quick and efficient purification of phyllacanthone from <i>C. quercifolius</i> crude extract with a yield of 4.5% (w/w) and a purity of 83%. Favelines-enriched fractions were also obtained and purified by preparative HPLC, resulting in the isolation of faveline, isofavelol, and deoxofaveline. In addition, phyllacanthone B, a previously unreported derivative, was identified and characterized. Pharmacological evaluations revealed the significant anti-melanoma activity of purified metabolites, with distinct structure–activity relationships. These results shed light on the mechanism of action of favelines, suggesting their potential as hits for the development of novel anticancer agents.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143404322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation of Urinary Extracellular Vesicles (EVs) via Hydrophobic Interaction Chromatography Using a Nylon-6 Capillary-Channeled Polymer (C-CP) Fiber Column","authors":"William F. Pons,&nbsp;R. Kenneth Marcus","doi":"10.1002/jssc.70093","DOIUrl":"https://doi.org/10.1002/jssc.70093","url":null,"abstract":"<p>Exosomes, a subset of extracellular vesicles (EVs) ranging in size from 30 to 150 nm, are of significant interest for biomedical applications such as diagnostic testing and therapeutics delivery. Biofluids, including urine, blood, and saliva, contain exosomes that carry biomarkers reflective of their host cells. However, isolation of EVs is often a challenge due to their size range, low density, and high hydrophobicity. Isolations can involve long separation times (ultracentrifugation) or result in impure eluates (size exclusion chromatography, polymer-based precipitation). As an alternative to these methods, this study evaluates the first use of nylon-6 capillary-channeled polymer (C-CP) fiber columns to separate EVs from human urine via a step-gradient hydrophobic interaction chromatography method. Different from previous efforts using polyester fiber columns for EV separations, nylon-6 shows potential for increased isolation efficiency, including somewhat higher column loading capacity and more gentle EV elution solvent strength. The efficacy of this approach to EV separation has been determined by scanning electron and transmission microscopy, nanoparticle flow cytometry (NanoFCM), and Bradford protein assays. Electron microscopy showed isolated vesicles of the expected morphology. Nanoparticle flow cytometry determined particle densities of eluates yielding up to 5 × 10⁸ particles mL⁻¹, a typical distribution of vesicle sizes in the eluate (60–100 nm), and immunoconfirmation using fluorescent anti-CD81 antibodies. Bradford assays confirmed that protein concentrations in the EV eluate were significantly reduced (approx. sevenfold) from raw urine. Overall, this approach provides a low-cost and time-efficient (&lt; 20 min) column separation to yield urinary EVs of the high purities required for downstream applications, including diagnostic testing and therapeutics.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70093","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389059","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation of Urinary Extracellular Vesicles (EVs) via Hydrophobic Interaction Chromatography Using a Nylon-6 Capillary-Channeled Polymer (C-CP) Fiber Column","authors":"William F. Pons,&nbsp;R. Kenneth Marcus","doi":"10.1002/jssc.70093","DOIUrl":"https://doi.org/10.1002/jssc.70093","url":null,"abstract":"<p>Exosomes, a subset of extracellular vesicles (EVs) ranging in size from 30 to 150 nm, are of significant interest for biomedical applications such as diagnostic testing and therapeutics delivery. Biofluids, including urine, blood, and saliva, contain exosomes that carry biomarkers reflective of their host cells. However, isolation of EVs is often a challenge due to their size range, low density, and high hydrophobicity. Isolations can involve long separation times (ultracentrifugation) or result in impure eluates (size exclusion chromatography, polymer-based precipitation). As an alternative to these methods, this study evaluates the first use of nylon-6 capillary-channeled polymer (C-CP) fiber columns to separate EVs from human urine via a step-gradient hydrophobic interaction chromatography method. Different from previous efforts using polyester fiber columns for EV separations, nylon-6 shows potential for increased isolation efficiency, including somewhat higher column loading capacity and more gentle EV elution solvent strength. The efficacy of this approach to EV separation has been determined by scanning electron and transmission microscopy, nanoparticle flow cytometry (NanoFCM), and Bradford protein assays. Electron microscopy showed isolated vesicles of the expected morphology. Nanoparticle flow cytometry determined particle densities of eluates yielding up to 5 × 10⁸ particles mL⁻¹, a typical distribution of vesicle sizes in the eluate (60–100 nm), and immunoconfirmation using fluorescent anti-CD81 antibodies. Bradford assays confirmed that protein concentrations in the EV eluate were significantly reduced (approx. sevenfold) from raw urine. Overall, this approach provides a low-cost and time-efficient (&lt; 20 min) column separation to yield urinary EVs of the high purities required for downstream applications, including diagnostic testing and therapeutics.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70093","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"First Dimension Trap-and-Elute Combined with Second Dimension Reversed-Phase Liquid Chromatography Separation Using a Two-Dimensional-Liquid Chromatography-Tandem Mass Spectrometry System for Sensitive Quantification of Human Insulin and Six Insulin Analogs in Plasma: Improved Chromatographic Resolution and Stability Testing","authors":"Pavel Sistik,&nbsp;Romana Urinovska,&nbsp;Klara Handlosova,&nbsp;Petr Handlos,&nbsp;Katerina Andelova,&nbsp;Jan Jurica,&nbsp;David Stejskal","doi":"10.1002/jssc.70092","DOIUrl":"https://doi.org/10.1002/jssc.70092","url":null,"abstract":"<p>Multidimensional chromatography coupled to tandem mass spectrometry (MS/MS), including simple sample preparation with protein precipitation, anion conversion with ammonium hydroxide, and solid-phase extraction using mixed-mode anion exchange in a 96-well plate format, has been validated for rapid simultaneous analysis of human insulin and its six analogs (lispro, glulisine, glargine, degludec, detemir, and aspart) in human plasma. This method is critical for clinical diagnostics, forensic investigations, and anti-doping efforts due to the widespread use of these substances. In the present study, improved chromatographic resolution was achieved using a first-dimension trap-and-elute configuration with an XBridge C18 (2.1 × 20 mm, 3.5 µm) trap column combined with second dimension separation on a Cortecs Ultra-High-Performance Liquid Chromatography (UHPLC) C18+ (2.1 × 100 mm, 1.6 µm) analytical column implemented within a two-dimensional-LC-MS/MS system. The total chromatographic run time was 11 min. This setup increases both the resolution and sensitivity of the method. A mobile phase consisting of 0.8% formic acid (FA) in water and 0.7% FA in acetonitrile was used for gradient elution. Bovine insulin was used as the internal standard. MS detection was performed in positive electrospray ionization mode, and the ion suppression due to matrix effects was evaluated. Validation criteria included linearity, precision, accuracy, recovery, lower limit of quantitation, matrix effect, and stability tests with and without protease inhibitor cocktail under different conditions (short-term stability, long-term stability, and freeze-thaw stability). The concentration range for all insulins was 50–15 000 pg/mL, with limits of quantification below the therapeutic reference range for all analytes. Intra-run precision ranged from 1.1% to 5.7%, inter-run precision from 0.7% to 5.9%, and overall recovery from 96.9% to 114.3%. The validated method has been implemented successfully by the Department of Forensic Medicine at our hospital for the investigation of unexplained deaths.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70092","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"First Dimension Trap-and-Elute Combined with Second Dimension Reversed-Phase Liquid Chromatography Separation Using a Two-Dimensional-Liquid Chromatography-Tandem Mass Spectrometry System for Sensitive Quantification of Human Insulin and Six Insulin Analogs in Plasma: Improved Chromatographic Resolution and Stability Testing","authors":"Pavel Sistik,&nbsp;Romana Urinovska,&nbsp;Klara Handlosova,&nbsp;Petr Handlos,&nbsp;Katerina Andelova,&nbsp;Jan Jurica,&nbsp;David Stejskal","doi":"10.1002/jssc.70092","DOIUrl":"https://doi.org/10.1002/jssc.70092","url":null,"abstract":"<p>Multidimensional chromatography coupled to tandem mass spectrometry (MS/MS), including simple sample preparation with protein precipitation, anion conversion with ammonium hydroxide, and solid-phase extraction using mixed-mode anion exchange in a 96-well plate format, has been validated for rapid simultaneous analysis of human insulin and its six analogs (lispro, glulisine, glargine, degludec, detemir, and aspart) in human plasma. This method is critical for clinical diagnostics, forensic investigations, and anti-doping efforts due to the widespread use of these substances. In the present study, improved chromatographic resolution was achieved using a first-dimension trap-and-elute configuration with an XBridge C18 (2.1 × 20 mm, 3.5 µm) trap column combined with second dimension separation on a Cortecs Ultra-High-Performance Liquid Chromatography (UHPLC) C18+ (2.1 × 100 mm, 1.6 µm) analytical column implemented within a two-dimensional-LC-MS/MS system. The total chromatographic run time was 11 min. This setup increases both the resolution and sensitivity of the method. A mobile phase consisting of 0.8% formic acid (FA) in water and 0.7% FA in acetonitrile was used for gradient elution. Bovine insulin was used as the internal standard. MS detection was performed in positive electrospray ionization mode, and the ion suppression due to matrix effects was evaluated. Validation criteria included linearity, precision, accuracy, recovery, lower limit of quantitation, matrix effect, and stability tests with and without protease inhibitor cocktail under different conditions (short-term stability, long-term stability, and freeze-thaw stability). The concentration range for all insulins was 50–15 000 pg/mL, with limits of quantification below the therapeutic reference range for all analytes. Intra-run precision ranged from 1.1% to 5.7%, inter-run precision from 0.7% to 5.9%, and overall recovery from 96.9% to 114.3%. The validated method has been implemented successfully by the Department of Forensic Medicine at our hospital for the investigation of unexplained deaths.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70092","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Protocol for Determination of Proteinogenic Amino Acids in Biological Fluids by the High-Speed UHPLC-MS Method: Application on Transgenic Spontaneously Hypertensive Rat-24 Plasma and Cerebrospinal Fluid Samples","authors":"Juraj Piestansky,&nbsp;Dominika Olesova,&nbsp;Petra Majerova,&nbsp;Petra Chalova,&nbsp;Andrej Kovac","doi":"10.1002/jssc.70089","DOIUrl":"https://doi.org/10.1002/jssc.70089","url":null,"abstract":"<div>\u0000 \u0000 <p>Recently, proteinogenic amino acids have become very interesting molecules, accompanied by a large variety of metabolic processes in humans and are associated with various diseases. In the era of system biology, including a broad spectrum of associated disciplines (e.g., metabolomics, lipidomics, proteomics, etc.), the possibility of identifying trustworthy biomarkers of diseases becomes much more likely. Changes in amino acid levels in plasma, serum, or cerebrospinal fluid reflect physiological or pathological conditions and, therefore, their regular monitoring can lead to early detection of the occurrence of a disease. Therefore, the exact determination of amino acids in biological fluids is of great importance. However, it is necessary to dispose with an effective, accurate, precise, selective, and robust analytical method. This protocol describes the complex procedure of amino acid analysis based on a combination of UHPLC with single quadrupole MS. The protocol presents a highly reproducible and robust methodology that has already been established in the quality control of biopharmaceuticals and determination of proteinogenic amino acids in urine in our laboratory. Here, the application potential is extended to the most frequently investigated biological fluid, that is, plasma and to the cerebrospinal fluid, which is investigated in many neurological conditions.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143248429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Protocol for Determination of Proteinogenic Amino Acids in Biological Fluids by the High-Speed UHPLC-MS Method: Application on Transgenic Spontaneously Hypertensive Rat-24 Plasma and Cerebrospinal Fluid Samples","authors":"Juraj Piestansky,&nbsp;Dominika Olesova,&nbsp;Petra Majerova,&nbsp;Petra Chalova,&nbsp;Andrej Kovac","doi":"10.1002/jssc.70089","DOIUrl":"https://doi.org/10.1002/jssc.70089","url":null,"abstract":"<div>\u0000 \u0000 <p>Recently, proteinogenic amino acids have become very interesting molecules, accompanied by a large variety of metabolic processes in humans and are associated with various diseases. In the era of system biology, including a broad spectrum of associated disciplines (e.g., metabolomics, lipidomics, proteomics, etc.), the possibility of identifying trustworthy biomarkers of diseases becomes much more likely. Changes in amino acid levels in plasma, serum, or cerebrospinal fluid reflect physiological or pathological conditions and, therefore, their regular monitoring can lead to early detection of the occurrence of a disease. Therefore, the exact determination of amino acids in biological fluids is of great importance. However, it is necessary to dispose with an effective, accurate, precise, selective, and robust analytical method. This protocol describes the complex procedure of amino acid analysis based on a combination of UHPLC with single quadrupole MS. The protocol presents a highly reproducible and robust methodology that has already been established in the quality control of biopharmaceuticals and determination of proteinogenic amino acids in urine in our laboratory. Here, the application potential is extended to the most frequently investigated biological fluid, that is, plasma and to the cerebrospinal fluid, which is investigated in many neurological conditions.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 2","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143248640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}