Development of a Quantitative LC-MS/MS-based Phenylalanine Hydroxylase Activity Assay

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July 17,2026

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Deficiency of PAH results in phenylketonuria (PKU), an inherited metabolic disorder characterized by elevated phenylalanine concentrations and progressive neurological impairment if untreated. Accurate measurement of hepatic PAH activity is essential for evaluating enzyme replacement and gene therapies. However, existing assays are indirect or require extensive sample preparation because of interference from endogenous tyrosine. This study aimed to develop a direct LC-MS/MS assay that quantifies tyrosine generated during an in vitro enzymatic reaction as a robust and quantitative measure of PAH activity in liver tissue.

Development of a Quantitative LC-MS/MS-based Phenylalanine Hydroxylase Activity Assay

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