A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s00216-018-1326-x http://hdl.handle.net/11449/176838 |
Resumo: | L-asparaginase or ASNase (L-asparagine aminohydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia (ALL) and lymphosarcoma through the depletion of L-asparagine (L-Asn) resulting in cytotoxicity to leukemic cells. ASNase is also important in the food industry, preventing acrylamide formation in processed foods. Several quantification techniques have been developed and used for the measurement of the ASNase activity, but standard pharmaceutical quality control methods were hardly reported, and in general, no official quality control guidelines were defined. To overcome this lack of information and to demonstrate the advantages and limitations, this work properly compares the traditional colorimetric methods (Nessler; L-aspartic acid β-hydroxamate (AHA); and indooxine) and the high-performance liquid chromatography (HPLC) method. A comparison of the methods using pure ASNase shows that the colorimetric methods both overestimate (Nessler) and underestimate (AHA and indooxine) the ASNase activity when compared to the values obtained with HPLC, considered the most precise method as this method monitors both substrate consumption and product formation, allowing for overall mass-balance. Correlation and critical analysis of each method relative to the HPLC method were carried out, resulting in a demonstration that it is crucial to select a proper method for the quantification of ASNase activity, allowing bioequivalence studies and individualized monitoring of different ASNase preparations. [Figure not available: see fulltext.] |
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A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatographyEnzymatic activityIndooxineL-asparaginaseL-aspartic acidL-aspartic acid β-hydroxamateNesslerL-asparaginase or ASNase (L-asparagine aminohydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia (ALL) and lymphosarcoma through the depletion of L-asparagine (L-Asn) resulting in cytotoxicity to leukemic cells. ASNase is also important in the food industry, preventing acrylamide formation in processed foods. Several quantification techniques have been developed and used for the measurement of the ASNase activity, but standard pharmaceutical quality control methods were hardly reported, and in general, no official quality control guidelines were defined. To overcome this lack of information and to demonstrate the advantages and limitations, this work properly compares the traditional colorimetric methods (Nessler; L-aspartic acid β-hydroxamate (AHA); and indooxine) and the high-performance liquid chromatography (HPLC) method. A comparison of the methods using pure ASNase shows that the colorimetric methods both overestimate (Nessler) and underestimate (AHA and indooxine) the ASNase activity when compared to the values obtained with HPLC, considered the most precise method as this method monitors both substrate consumption and product formation, allowing for overall mass-balance. Correlation and critical analysis of each method relative to the HPLC method were carried out, resulting in a demonstration that it is crucial to select a proper method for the quantification of ASNase activity, allowing bioequivalence studies and individualized monitoring of different ASNase preparations. [Figure not available: see fulltext.]Department of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01, Campos VilleBiochemistry and Technology Chemistry Department Chemistry Institute São Paulo State University (UNESP)Department of Chemistry Earlham College, 801 National Road WestDepartment of Biochemical-Pharmaceutical Technology Pharmaceutical Biotechnology Laboratory University of Sao Paulo (USP)Department of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01, Campos VilleBiochemistry and Technology Chemistry Department Chemistry Institute São Paulo State University (UNESP)Universidade Estadual Paulista (Unesp)Earlham CollegeUniversidade de São Paulo (USP)Magri, Agnes [UNESP]Soler, Matheus F. [UNESP]Lopes, André M. [UNESP]Cilli, Eduardo M. [UNESP]Barber, Patrick S.Pessoa, AdalbertoPereira, Jorge F. B. [UNESP]2018-12-11T17:22:42Z2018-12-11T17:22:42Z2018-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1007/s00216-018-1326-xAnalytical and Bioanalytical Chemistry.1618-26501618-2642http://hdl.handle.net/11449/17683810.1007/s00216-018-1326-x2-s2.0-850532637172-s2.0-85053263717.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengAnalytical and Bioanalytical Chemistry0,9780,978info:eu-repo/semantics/openAccess2023-11-16T06:10:57Zoai:repositorio.unesp.br:11449/176838Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:50:52.136262Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
title |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
spellingShingle |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography Magri, Agnes [UNESP] Enzymatic activity Indooxine L-asparaginase L-aspartic acid L-aspartic acid β-hydroxamate Nessler |
title_short |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
title_full |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
title_fullStr |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
title_full_unstemmed |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
title_sort |
A critical analysis of L-asparaginase activity quantification methods—colorimetric methods versus high-performance liquid chromatography |
author |
Magri, Agnes [UNESP] |
author_facet |
Magri, Agnes [UNESP] Soler, Matheus F. [UNESP] Lopes, André M. [UNESP] Cilli, Eduardo M. [UNESP] Barber, Patrick S. Pessoa, Adalberto Pereira, Jorge F. B. [UNESP] |
author_role |
author |
author2 |
Soler, Matheus F. [UNESP] Lopes, André M. [UNESP] Cilli, Eduardo M. [UNESP] Barber, Patrick S. Pessoa, Adalberto Pereira, Jorge F. B. [UNESP] |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Earlham College Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Magri, Agnes [UNESP] Soler, Matheus F. [UNESP] Lopes, André M. [UNESP] Cilli, Eduardo M. [UNESP] Barber, Patrick S. Pessoa, Adalberto Pereira, Jorge F. B. [UNESP] |
dc.subject.por.fl_str_mv |
Enzymatic activity Indooxine L-asparaginase L-aspartic acid L-aspartic acid β-hydroxamate Nessler |
topic |
Enzymatic activity Indooxine L-asparaginase L-aspartic acid L-aspartic acid β-hydroxamate Nessler |
description |
L-asparaginase or ASNase (L-asparagine aminohydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia (ALL) and lymphosarcoma through the depletion of L-asparagine (L-Asn) resulting in cytotoxicity to leukemic cells. ASNase is also important in the food industry, preventing acrylamide formation in processed foods. Several quantification techniques have been developed and used for the measurement of the ASNase activity, but standard pharmaceutical quality control methods were hardly reported, and in general, no official quality control guidelines were defined. To overcome this lack of information and to demonstrate the advantages and limitations, this work properly compares the traditional colorimetric methods (Nessler; L-aspartic acid β-hydroxamate (AHA); and indooxine) and the high-performance liquid chromatography (HPLC) method. A comparison of the methods using pure ASNase shows that the colorimetric methods both overestimate (Nessler) and underestimate (AHA and indooxine) the ASNase activity when compared to the values obtained with HPLC, considered the most precise method as this method monitors both substrate consumption and product formation, allowing for overall mass-balance. Correlation and critical analysis of each method relative to the HPLC method were carried out, resulting in a demonstration that it is crucial to select a proper method for the quantification of ASNase activity, allowing bioequivalence studies and individualized monitoring of different ASNase preparations. [Figure not available: see fulltext.] |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-11T17:22:42Z 2018-12-11T17:22:42Z 2018-01-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s00216-018-1326-x Analytical and Bioanalytical Chemistry. 1618-2650 1618-2642 http://hdl.handle.net/11449/176838 10.1007/s00216-018-1326-x 2-s2.0-85053263717 2-s2.0-85053263717.pdf |
url |
http://dx.doi.org/10.1007/s00216-018-1326-x http://hdl.handle.net/11449/176838 |
identifier_str_mv |
Analytical and Bioanalytical Chemistry. 1618-2650 1618-2642 10.1007/s00216-018-1326-x 2-s2.0-85053263717 2-s2.0-85053263717.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Analytical and Bioanalytical Chemistry 0,978 0,978 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808128865875263488 |