Determination of free trimethyllysine in plasma and tissue specimens by high-performance liquid chromatography

A method for the determination of 6-N-trimethyllysine in tissues and plasma is described. Trimethyllysine and the chemically analogous 6-N-triethyllysine (internal standard) were isolated from acid-soluble fractions of tissue homogenates or plasma by combined ion-exchange--ion-exclusion chromatography. Trimethyllysine and triethyllysine were separated from other sample constituents by reversed-phase ion-pair high-performance liquid chromatography, derivatized post-column by reaction with o-phthalicdicarboxaldehyde and 2-mercaptoethanol, and detected fluorometrically. Standard curves were linear over a sample concentration range of 0.5--4 nmol/ml. The detection limit corresponded with 25 pmol trimethyllysine injected into the chromatograph. The procedure was used for the determination of trimethyllysine in plasma, liver, kidney, and mixed skeletal muscle of rat.