강유진 (센트럴바이오)

장지훈 (센트럴바이오)

문주연 (센트럴바이오)

강유진 (센트럴바이오)

하나리 (유한양행)

이보람 (유한양행)

안수진 (유한양행)

장지훈 (센트럴바이오)

문주연 (센트럴바이오)

Methylamine is an endogenous metabolite of clinical and experimental interest, as it has been implicated in various pathological conditions including renal dysfunction, lysosomal storage disorders, and certain cancers. However, due to its high polarity and low molecular weight, the quantitative analysis of endogenous methylamine in biological matrices by LC-MS/MS remains highly challenging. To achieve accurate and reliable quantification, a comprehensive analytical approach is essential.

We developed and optimized an LC-MS/MS-based method involving chemical derivatization of methylamine’s amine group with tert-butyl bromoacetate (TBBA) under alkaline conditions. Key reaction parameters- concentration, temperature, and reaction time-were systematically optimized to enhance derivatization efficiency. The resulting methylamine-TBBA derivative exhibited increased lipophilicity, significantly improving chromatographic retention and ionization efficiency in LC-MS/MS analysis. The method demonstrated excellent linearity (r >​ 0.999) over the concentration range of 20 to 1,500 ng/mL, with accuracy (%Bias) ranging from -5.7% to 5.6%. The method was successfully applied to quantify methylamine in mouse plasma samples, demonstrating its applicability to real biological matrices.

This robust and sensitive technique offers a valuable tool for the quantitative analysis of methylamine and holds promise as an endogenous metabolite analysis in studies of related pathological conditions.