A B S T R A C T
Advancements in liquid chromatography-tandem mass spectrometry (LC-MS/MS) are redefining the landscape of
clinical diagnostics, particularly in the context of newborn screening for inborn errors of metabolism. Conventional
analytical platforms are often limited by a small number of analytes and require multiple platforms for
subsequent analyses, thereby impacting timely diagnosis and management. This study describes the development
of a multiplexed targeted mass spectrometry-based assay for simultaneous detection and quantitation of amines,
including amino acids and their derivatives, from plasma, urine and dried blood spots. The method utilizes a
single step derivatization strategy based on one step click chemistry that simplifies sample preparation while
improving analytical sensitivity allowing for detection of analytes at sub-picomolar concentrations. Furthermore,
we adopted a strategy of generating heavy isotopically labeled standards by chemically modifying the corresponding
light standards using a stable isotope labeled derivatizing agent, enabling their use as internal standards
for quantification. This approach can offer a cost-effective and scalable solution for the early detection and
management of inherited metabolic disorders, particularly in cases where accurate detection of amines is critical.
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