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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
2009-1-15
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pubmed:abstractText |
A novel assay for the determination of l-asparaginase activity in human plasma is described that is based on the HPLC quantitation of l-aspartic acid produced during enzyme incubation. Methods for monitoring l-asparagine depletion are also described. Chromatography of l-aspartic acid, l-asparagine and l-homoserine (the internal standard) involved derivatization with o-pthaldialdehyde, then separation from other amino acids on a Phenomenex Luna C(18) column using a 1 mL/min flow rate and a mobile phase consisting of di-potassium hydrogen orthophosphate propionate buffer, pH 6, with 10% methanol and 10% acetonitrile. Fluoresence detection was at excitation/emission wavelengths of 357/455 nm. Under these conditions l-aspartic acid, l-asparagine and l-homoserine had retention times of 3.5, 9.8 and 17.7 min, respectively. The l-asparaginase assay was linear from 0.1 to 10 U/mL activity and interday precision and accuracy were less than 13%. The limit of quantitation was approximately 0.03 U/mL. The assay utility was established in 12 children who received E. coli l-asparaginase as treatment for acute lymphoblastic leukaemia.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
1099-0801
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pubmed:author |
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pubmed:copyrightInfo |
Copyright (c) 2008 John Wiley & Sons, Ltd.
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pubmed:issnType |
Electronic
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pubmed:volume |
23
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
152-9
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pubmed:dateRevised |
2009-7-17
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pubmed:meshHeading |
pubmed-meshheading:18823071-Adolescent,
pubmed-meshheading:18823071-Antineoplastic Agents,
pubmed-meshheading:18823071-Asparaginase,
pubmed-meshheading:18823071-Asparagine,
pubmed-meshheading:18823071-Aspartic Acid,
pubmed-meshheading:18823071-Child,
pubmed-meshheading:18823071-Child, Preschool,
pubmed-meshheading:18823071-Chromatography, High Pressure Liquid,
pubmed-meshheading:18823071-Drug Stability,
pubmed-meshheading:18823071-Escherichia coli Proteins,
pubmed-meshheading:18823071-Fluorescence,
pubmed-meshheading:18823071-Glutamic Acid,
pubmed-meshheading:18823071-Glutamine,
pubmed-meshheading:18823071-Homoserine,
pubmed-meshheading:18823071-Humans,
pubmed-meshheading:18823071-Least-Squares Analysis,
pubmed-meshheading:18823071-Precursor Cell Lymphoblastic Leukemia-Lymphoma,
pubmed-meshheading:18823071-Reference Standards,
pubmed-meshheading:18823071-Reproducibility of Results,
pubmed-meshheading:18823071-Sensitivity and Specificity
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pubmed:year |
2009
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pubmed:articleTitle |
An isocratic fluorescence HPLC assay for the monitoring of l-asparaginase activity and l-asparagine depletion in children receiving E. colil-asparaginase for the treatment of acute lymphoblastic leukaemia.
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pubmed:affiliation |
Department of Biochemistry, The Children's Hospital at Westmead, Westmead, NSW, Australia. christan@chw.edu.au
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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