5'- Nucleotidase (5'-NT) Assay

Assay Principle
The 5'-NT assay is based on the enzymatic hydrolysis of 5'-monophosphate (5'-IMP) to form inosine which is converted to hypoxanthine by purine nucleoside phosphorylase (PNP). Hypoxanthine is then converted to uric acid and hydrogen peroxide (H2O2) by xanthine oxidase (XOD). H2O2 is further reacted with N-Ethyl- N-(2-hydroxy-3-sulfopropyl)-3-methylaniline (EHSPT) and 4- aminoantipyrine (4-AA) in the presence of peroxidase (POD) to generate quinone dye which is monitored in a kinetic manner.

Features
5'-Nucleotidase Assay is linear from 0 - 300 U/L (R² >0.99) with intra-assay CV% of < 2% and an inter-assay CV% of ≤ 4%. 5'-Nucleotidase assay is not affected by interfering substances such as serum bilirubin up to 40 mg/dL, hemoglobin up to 500 mg/dL, triglycerides up to 1250 mg/dL, ascorbic acid up to 20 mg/dL, and alkaline phosphatase up to 1250 U/L.

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