Effect of pH on spectral characteristics of P5C-ninhydrin derivative: Application in the assay of ornithine amino transferase activity from tissue lysate |
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| Authors: | H Ravikumar K S Devaraju K Taranath Shetty |
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| Institution: | (1) Department of Neurochemistry, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, Karnataka, India;(2) Department of Biotechnology, Bangalore University, Bangalore, 560056, India |
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| Abstract: | Currently available method(s) for assaying pyrroline-5-carboxylate (P5C), an important intermediate metabolite of ornithine,
proline and glutamate metabolic pathways, are cumbersome or not sensitive enough for microanalysis. The present study involving
the synthesis of P5C followed by purity check, molecular mass (amu =113.1) determination by mass spectrometry and spectral
characterization of P5C-ninhydrin derivative (λ max: 510 nm) confirmed the authenticity of the preparation. Studies on the
effect of pH on spectral characteristics of P5C ninhydrin derivative demonstrated a significant change with respect to λ max
(620 nm) and several ∼ 12 fold increase in molar extinction coefficient (ε: 1.96 × 105) in alkaline conditions (pH:7.0–8.0) as compared to the reported Molar ε of 1.65 × 104 at max λ 510 nm in ethanolic solution. The modified method, with the improved sensitivity, is adopted for the assay of ornithine
amino transferase activity in WBC’s/platelets lysate(s) from human blood. |
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| Keywords: | Pyrroline-5-Carboxylate Mass spectrometry WBC’ s/platelets lysate(s) Ornithine amino-transferase Pyrroline-5-Carboxylate-Ninhydrin complex |
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