Fluorometric micromethod for determination of arginase activity in dried blood spots on filter paper

We describe a microfluorometric method for determination of arginase activity in dried blood spots on filter paper. The arginase in discs punched from such dried blood specimens is activated by preincubation with Mn2+ at 37 degrees C. After incubation with substrate at the same temperature, urea is...

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Bibliographic Details
Published inClinical chemistry (Baltimore, Md.) Vol. 26; no. 8; pp. 1198 - 1200
Main Authors Orfanos, AP, Naylor, EW, Guthrie, R
Format Journal Article
LanguageEnglish
Published England Am Assoc Clin Chem 01.07.1980
Subjects
Adult
Arginase - blood
Blood Specimen Collection
Clinical Enzyme Tests
Drug Stability
Hematologic Diseases - diagnosis
Hematologic Diseases - genetics
Humans
Infant, Newborn
Infant, Newborn, Diseases - diagnosis
Reference Values
Spectrometry, Fluorescence - methods
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Summary:We describe a microfluorometric method for determination of arginase activity in dried blood spots on filter paper. The arginase in discs punched from such dried blood specimens is activated by preincubation with Mn2+ at 37 degrees C. After incubation with substrate at the same temperature, urea is determined fluorometrically by oxidation of NADH to NAD+ in a coupled kinetic reaction. We compare the results of this method with those of a colorimetric method involving liquid blood samples, and assess the stability of the enzyme in dried blood on filter paper. The presence of serum has no effect on the activity. This method may be useful in the early detection of arginase deficiency and certain hematological disorders.
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ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/26.8.1198