Assay of Ornithine Aminotransferase with Ninhydrin

We developed an assay system for ornithine amino-transferase (EC 2.6.1.13) using ninhydrin. Pyrroline 5-carboxylate, a product of enzymatic transamination, reacts with ninhydrin under hot acidic conditions to form a reddish pigment soluble in ethanol. The millimolar extinction coefficient of reactio...

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Bibliographic Details
Published in:Analytical biochemistry 1994-12, Vol.223 (2), p.205-207
Main Authors: Kim, H.R., Rho, H.W., Park, J.W., Park, B.H., Kim, J.S., Lee, M.W.
Format: Article
Language:English
Subjects:
Animals
Benzaldehydes
Colorimetry
Drug Stability
Intestine, Small - enzymology
Liver - enzymology
Male
Mice
Mice, Inbred ICR
Ninhydrin
Ornithine-Oxo-Acid Transaminase - analysis
Pyrroles
Sensitivity and Specificity
Spectrophotometry
Tissue Distribution
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Summary:We developed an assay system for ornithine amino-transferase (EC 2.6.1.13) using ninhydrin. Pyrroline 5-carboxylate, a product of enzymatic transamination, reacts with ninhydrin under hot acidic conditions to form a reddish pigment soluble in ethanol. The millimolar extinction coefficient of reaction product dissolved in ethanol was 16.5 at 510 nm. Acidification with perchloric acid effectively abolished the interfering color development by L-ornithine and L-glutamate. The paired activity measurement in mouse tissues by ninhydrin and o-aminobenzaldehyde methods showed a good correlation (γ = 0.985). In our ninhydrin method, stable ninhydrin replaced unstable o-aminobenzaldehyde, and sensitivity was much higher than that with the conventional o-aminobenzaldehyde method.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1994.1574