Journal of separation science最新文献_第6页

From Complexity to Clarity: Expanding Metabolome Coverage With Innovative Analytical Strategies

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70099

Kanukolanu Aarika, Ramijinni Rajyalakshmi, Lakshmi Vineela Nalla, Siva Nageswara Rao Gajula

{"title":"From Complexity to Clarity: Expanding Metabolome Coverage With Innovative Analytical Strategies","authors":"Kanukolanu Aarika, Ramijinni Rajyalakshmi, Lakshmi Vineela Nalla, Siva Nageswara Rao Gajula","doi":"10.1002/jssc.70099","DOIUrl":"https://doi.org/10.1002/jssc.70099","url":null,"abstract":"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.","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":"143438957","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}

引用次数: 0

Development of a Catalyst-Free Ultrasound-Assisted Derivatization Method for Detection of Valproic Acid in Epilepsy Patient's Serum Using High-Performance Liquid Chromatography: A Comparison With Chemiluminescence Immunoassay

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70097

Javad Jaberi, Nikoo Alavi, Milad Moayednia

{"title":"Development of a Catalyst-Free Ultrasound-Assisted Derivatization Method for Detection of Valproic Acid in Epilepsy Patient's Serum Using High-Performance Liquid Chromatography: A Comparison With Chemiluminescence Immunoassay","authors":"Javad Jaberi, Nikoo Alavi, Milad Moayednia","doi":"10.1002/jssc.70097","DOIUrl":"https://doi.org/10.1002/jssc.70097","url":null,"abstract":"<div>\u0000 \u0000 The aim of this research was the therapeutic drug monitoring for valproic acid in epilepsy patient's serum samples by the common, sensitive, and accessible HPLC-UV method. Because of the absence of a suitable chromophore in the valproic acid structure, a facile, selective, and cost-effective pre-column derivatization was designed. This catalyst-free ultrasound-assisted derivatization assay can accomplish the derivatization very quickly only in 5.0 min and at a mild temperature of 60°C. 2,4ʹ-Dibromoacetophenone and nonanoic acid was used as derivatizing agent and internal standard, respectively. The effect of sample pH, buffer concentration, ultrasound exposure time, reaction temperature, and derivatizing agent amount were optimized. The proposed method exhibited a good linear range of 5.0–300.0 µg/mL with acceptable correlation coefficients of 0.9981. The limit of detection was as low as 0.4 µg/mL. Also, the limit of quantification was reported as 1.3 µg/mL. Interday and intraday relative standard deviations (n = 10) were 1.1% and 0.3%–7.0%, respectively. In addition, the relative recovery ranged from 100.3% to 107.7%. The measurement of valproic acid was performed in the presence of several epilepsy and non-epilepsy drugs by the developed protocol. This confirmed the specific and accurate determination of valproic acid in the patient's serum. A comparative evaluation was employed against the precise chemiluminescence immunoassay approach. The correlation coefficient between the two methods was 0.9992, which demonstrated the results were statistically the same.\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":"143439214","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}

引用次数: 0

Development of a Catalyst-Free Ultrasound-Assisted Derivatization Method for Detection of Valproic Acid in Epilepsy Patient's Serum Using High-Performance Liquid Chromatography: A Comparison With Chemiluminescence Immunoassay

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70097

Javad Jaberi, Nikoo Alavi, Milad Moayednia

{"title":"Development of a Catalyst-Free Ultrasound-Assisted Derivatization Method for Detection of Valproic Acid in Epilepsy Patient's Serum Using High-Performance Liquid Chromatography: A Comparison With Chemiluminescence Immunoassay","authors":"Javad Jaberi, Nikoo Alavi, Milad Moayednia","doi":"10.1002/jssc.70097","DOIUrl":"https://doi.org/10.1002/jssc.70097","url":null,"abstract":"<div>\u0000 \u0000 The aim of this research was the therapeutic drug monitoring for valproic acid in epilepsy patient's serum samples by the common, sensitive, and accessible HPLC-UV method. Because of the absence of a suitable chromophore in the valproic acid structure, a facile, selective, and cost-effective pre-column derivatization was designed. This catalyst-free ultrasound-assisted derivatization assay can accomplish the derivatization very quickly only in 5.0 min and at a mild temperature of 60°C. 2,4ʹ-Dibromoacetophenone and nonanoic acid was used as derivatizing agent and internal standard, respectively. The effect of sample pH, buffer concentration, ultrasound exposure time, reaction temperature, and derivatizing agent amount were optimized. The proposed method exhibited a good linear range of 5.0–300.0 µg/mL with acceptable correlation coefficients of 0.9981. The limit of detection was as low as 0.4 µg/mL. Also, the limit of quantification was reported as 1.3 µg/mL. Interday and intraday relative standard deviations (n = 10) were 1.1% and 0.3%–7.0%, respectively. In addition, the relative recovery ranged from 100.3% to 107.7%. The measurement of valproic acid was performed in the presence of several epilepsy and non-epilepsy drugs by the developed protocol. This confirmed the specific and accurate determination of valproic acid in the patient's serum. A comparative evaluation was employed against the precise chemiluminescence immunoassay approach. The correlation coefficient between the two methods was 0.9992, which demonstrated the results were statistically the same.\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":"143439220","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}

引用次数: 0

Chiral Separation and Determination of Multiple Organophosphorus Pesticide Enantiomers in Soil Based on Cellulose-Based Chiral Column by LC–MS/MS

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70100

Liang Li, Rendan Zhou, Huiying Xie, Gang Li, Zhiguang Xu, Meijia Liu, Wanting Gao

{"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, Rendan Zhou, Huiying Xie, Gang Li, Zhiguang Xu, Meijia Liu, Wanting Gao","doi":"10.1002/jssc.70100","DOIUrl":"https://doi.org/10.1002/jssc.70100","url":null,"abstract":"<div>\u0000 \u0000 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 (Rs) 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 (R > 0.998), accuracy with recoveries ranging from 79% to 121%, precision with RSD% < 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.\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":"143439211","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}

引用次数: 0

Chiral Separation and Determination of Multiple Organophosphorus Pesticide Enantiomers in Soil Based on Cellulose-Based Chiral Column by LC–MS/MS

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70100

Liang Li, Rendan Zhou, Huiying Xie, Gang Li, Zhiguang Xu, Meijia Liu, Wanting Gao

{"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, Rendan Zhou, Huiying Xie, Gang Li, Zhiguang Xu, Meijia Liu, Wanting Gao","doi":"10.1002/jssc.70100","DOIUrl":"https://doi.org/10.1002/jssc.70100","url":null,"abstract":"<div>\u0000 \u0000 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 (Rs) 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 (R > 0.998), accuracy with recoveries ranging from 79% to 121%, precision with RSD% < 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.\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}

引用次数: 0

From Complexity to Clarity: Expanding Metabolome Coverage With Innovative Analytical Strategies

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-19 DOI: 10.1002/jssc.70099

Kanukolanu Aarika, Ramijinni Rajyalakshmi, Lakshmi Vineela Nalla, Siva Nageswara Rao Gajula

{"title":"From Complexity to Clarity: Expanding Metabolome Coverage With Innovative Analytical Strategies","authors":"Kanukolanu Aarika, Ramijinni Rajyalakshmi, Lakshmi Vineela Nalla, Siva Nageswara Rao Gajula","doi":"10.1002/jssc.70099","DOIUrl":"https://doi.org/10.1002/jssc.70099","url":null,"abstract":"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.","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}

引用次数: 0

Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-13 DOI: 10.1002/jssc.70094

Sovannychloé Nai, Christiane Adrielly Alves Ferraz, Thomas Gaslonde, Jackson Roberto Guedes da Silva Almeida, Edilson Beserra de Alencar Filho, Cíntia Emi Yanaguibashi Leal, Benjamin Musnier, Laurent Picot, Pierre-Edouard Bodet, Marie-Christine Lallemand, Raphaël Grougnet, Raimundo Gonçalves de Oliveira Junior

{"title":"Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography","authors":"Sovannychloé Nai, Christiane Adrielly Alves Ferraz, Thomas Gaslonde, Jackson Roberto Guedes da Silva Almeida, Edilson Beserra de Alencar Filho, Cíntia Emi Yanaguibashi Leal, Benjamin Musnier, Laurent Picot, Pierre-Edouard Bodet, Marie-Christine Lallemand, Raphaël Grougnet, Raimundo Gonçalves de Oliveira Junior","doi":"10.1002/jssc.70094","DOIUrl":"https://doi.org/10.1002/jssc.70094","url":null,"abstract":"Favelines, benzocycloheptene diterpenes exclusively found in Cnidoscolus quercifolius, 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 C. quercifolius 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.","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}

引用次数: 0

Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-13 DOI: 10.1002/jssc.70094

Sovannychloé Nai, Christiane Adrielly Alves Ferraz, Thomas Gaslonde, Jackson Roberto Guedes da Silva Almeida, Edilson Beserra de Alencar Filho, Cíntia Emi Yanaguibashi Leal, Benjamin Musnier, Laurent Picot, Pierre-Edouard Bodet, Marie-Christine Lallemand, Raphaël Grougnet, Raimundo Gonçalves de Oliveira Junior

{"title":"Single-Step Isolation of Phyllacanthone From Cnidoscolus quercifolius Using Centrifugal Partition Chromatography","authors":"Sovannychloé Nai, Christiane Adrielly Alves Ferraz, Thomas Gaslonde, Jackson Roberto Guedes da Silva Almeida, Edilson Beserra de Alencar Filho, Cíntia Emi Yanaguibashi Leal, Benjamin Musnier, Laurent Picot, Pierre-Edouard Bodet, Marie-Christine Lallemand, Raphaël Grougnet, Raimundo Gonçalves de Oliveira Junior","doi":"10.1002/jssc.70094","DOIUrl":"https://doi.org/10.1002/jssc.70094","url":null,"abstract":"Favelines, benzocycloheptene diterpenes exclusively found in Cnidoscolus quercifolius, 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 C. quercifolius 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.","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}

引用次数: 0

Isolation of Urinary Extracellular Vesicles (EVs) via Hydrophobic Interaction Chromatography Using a Nylon-6 Capillary-Channeled Polymer (C-CP) Fiber Column

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-11 DOI: 10.1002/jssc.70093

William F. Pons, R. Kenneth Marcus

{"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, R. Kenneth Marcus","doi":"10.1002/jssc.70093","DOIUrl":"https://doi.org/10.1002/jssc.70093","url":null,"abstract":"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 (< 20 min) column separation to yield urinary EVs of the high purities required for downstream applications, including diagnostic testing and therapeutics.","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}

引用次数: 0

Isolation of Urinary Extracellular Vesicles (EVs) via Hydrophobic Interaction Chromatography Using a Nylon-6 Capillary-Channeled Polymer (C-CP) Fiber Column

IF 2.8 3区工程技术

Journal of separation science Pub Date : 2025-02-11 DOI: 10.1002/jssc.70093

William F. Pons, R. Kenneth Marcus

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引用次数: 0