Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques

Background: In recent times, various human health disorders including cancer, diabetes, neurodegenerative, and metabolic diseases are noticed among human populations. Currently, genetic and proteomic approaches are highly reported to detect metabolic disorders that also include inborn error of metab...

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Main Authors: Ajay Kumar, Jainish Kothari, Devyani Bhatkar, Manmohan Mitruka, Roshni Pal, Sachin C Sarode, Nilesh Kumar Sharma
Format: Article
Language:English
Published: Wolters Kluwer Medknow Publications 2020-01-01
Series:Journal of Dental Research and Review
Subjects:
Online Access:http://www.jdrr.org/article.asp?issn=2348-2915;year=2020;volume=7;issue=5;spage=36;epage=40;aulast=Kumar
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spelling doaj-a3b81377f83a45ab9195f7419ba9cc4c2020-11-25T01:39:09ZengWolters Kluwer Medknow PublicationsJournal of Dental Research and Review2348-29152348-31722020-01-0175364010.4103/jdrr.jdrr_65_19Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniquesAjay KumarJainish KothariDevyani BhatkarManmohan MitrukaRoshni PalSachin C SarodeNilesh Kumar SharmaBackground: In recent times, various human health disorders including cancer, diabetes, neurodegenerative, and metabolic diseases are noticed among human populations. Currently, genetic and proteomic approaches are highly reported to detect metabolic disorders that also include inborn error of metabolisms. These existing detection methods are faced with cost issue and time-consuming factors. Therefore, metabolites as biomarkers are one of the potential avenues to detect metabolic disorders. Further, exploitation of urine as potential source of metabolite biomarkers, there are limitations in this area of research due to abundance of nonmetabolite components such as proteins and nucleic acids. Hence, methods and processes are required to precisely fractionate metabolites from urine of inborn error of metabolism patients and are then identified by analytical tools such as liquid chromatography–high-resolution mass spectrometry (LC-HRMS) and gas chromatography-tandem mass spectrometry. Materials and Methods: Sterile filtered urine samples (750 μl) mixed with (250 μl) loading buffer were electrophoresed on VTGE that uses acrylamide gel (acrylamide: bisacrylamide, 30:1) as a matrix of 15%. Further, vertical tube gel electrophoresis (VTGE) technique combined with LC-HRMS to identify metabolites that are known as the biomarkers of metabolic disorders was carried out. Results and Discussion: The authors provide evidence on the use of novel VTGE coupled with LC-HRMS to detect metabolites among metabolic disorders. Data suggest the applicability of VTGE coupled with LC-HRMS technique to detect metabolites such as 2-methyluridine, 2-methylglutaric acid, 2-methylcitric acid, and 2-hydroxyglutaric acid in case of metabolic disorders. Conclusion: This preliminary work is suggested to be extended to large clinical samples to validate application of this method to detect metabolic disorders including the inborn error of metabolisms.http://www.jdrr.org/article.asp?issn=2348-2915;year=2020;volume=7;issue=5;spage=36;epage=40;aulast=Kumarbiomarkersmass spectrometrymetabolic disordersmetabolitesurine
collection DOAJ
language English
format Article
sources DOAJ
author Ajay Kumar
Jainish Kothari
Devyani Bhatkar
Manmohan Mitruka
Roshni Pal
Sachin C Sarode
Nilesh Kumar Sharma
spellingShingle Ajay Kumar
Jainish Kothari
Devyani Bhatkar
Manmohan Mitruka
Roshni Pal
Sachin C Sarode
Nilesh Kumar Sharma
Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
Journal of Dental Research and Review
biomarkers
mass spectrometry
metabolic disorders
metabolites
urine
author_facet Ajay Kumar
Jainish Kothari
Devyani Bhatkar
Manmohan Mitruka
Roshni Pal
Sachin C Sarode
Nilesh Kumar Sharma
author_sort Ajay Kumar
title Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
title_short Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
title_full Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
title_fullStr Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
title_full_unstemmed Detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
title_sort detection of urinary metabolites of metabolic pathway disorders using vertical tube gel electrophoresis and liquid chromatography–high-resolution mass spectrometry techniques
publisher Wolters Kluwer Medknow Publications
series Journal of Dental Research and Review
issn 2348-2915
2348-3172
publishDate 2020-01-01
description Background: In recent times, various human health disorders including cancer, diabetes, neurodegenerative, and metabolic diseases are noticed among human populations. Currently, genetic and proteomic approaches are highly reported to detect metabolic disorders that also include inborn error of metabolisms. These existing detection methods are faced with cost issue and time-consuming factors. Therefore, metabolites as biomarkers are one of the potential avenues to detect metabolic disorders. Further, exploitation of urine as potential source of metabolite biomarkers, there are limitations in this area of research due to abundance of nonmetabolite components such as proteins and nucleic acids. Hence, methods and processes are required to precisely fractionate metabolites from urine of inborn error of metabolism patients and are then identified by analytical tools such as liquid chromatography–high-resolution mass spectrometry (LC-HRMS) and gas chromatography-tandem mass spectrometry. Materials and Methods: Sterile filtered urine samples (750 μl) mixed with (250 μl) loading buffer were electrophoresed on VTGE that uses acrylamide gel (acrylamide: bisacrylamide, 30:1) as a matrix of 15%. Further, vertical tube gel electrophoresis (VTGE) technique combined with LC-HRMS to identify metabolites that are known as the biomarkers of metabolic disorders was carried out. Results and Discussion: The authors provide evidence on the use of novel VTGE coupled with LC-HRMS to detect metabolites among metabolic disorders. Data suggest the applicability of VTGE coupled with LC-HRMS technique to detect metabolites such as 2-methyluridine, 2-methylglutaric acid, 2-methylcitric acid, and 2-hydroxyglutaric acid in case of metabolic disorders. Conclusion: This preliminary work is suggested to be extended to large clinical samples to validate application of this method to detect metabolic disorders including the inborn error of metabolisms.
topic biomarkers
mass spectrometry
metabolic disorders
metabolites
urine
url http://www.jdrr.org/article.asp?issn=2348-2915;year=2020;volume=7;issue=5;spage=36;epage=40;aulast=Kumar
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