A fluorometric high-performance liquid chromatography procedure for simultaneous determination of methylamine and aminoacetone in blood and tissues

Methylamine and aminoacetone are endogenous aliphatic amines found in human blood and urine. They can be oxidized by semicarbazide-sensitive amine oxidase (SSAO), leading to the production of toxic aldehydes such as formaldehyde and methylglyoxal as well as hydrogen peroxide and ammonia. SSAO is localized on the surface of vascular endothelial and smooth muscle cells and of adipocytes. Increases in SSAO activity are linked to vascular disorders associated with pathological conditions such as diabetic complications, heart failure, and vascular dementia. Quantitative assessment of methylamine and acetonitrile in tissues has been hampered due to the volatility and hydrolipophilicity of these amines as well as interference by complex biological constituents. We have overcome this problem and developed an FMOC/HPLC (9-fluorenylmethyl chloroformate-Cl/high-performance liquid chromatography) method for simultaneous assessment of methylamine and aminoacetone. This method has been validated using rodent tissues with a detection limit at the picogram level. Methylamine and aminoacetone distributed unevenly among different tissues ranged from 0.1 to 27 nmol/g. To our knowledge, this is the first report on simultaneous determination of methylamine and aminoacetone in mammal tissues.