Determination of 3-methylhistidine and 1-methylhistidine in untreated urine samples by capillary electrophoresis
Abstrakt
Capillary electrophoretic (CE) method was developed for the determination of urinary 3-methylhistidine (3MH) and 1-methylhistidine (1MH) indicating the extent of degradation of skeletal muscle proteins and thereby the state of human health. 3MH, 1MH and histidine can be separated in both acidic and alkaline media, where these amino acids form cation and anion, respectively. The effective mobility of all ionic forms was measured over a broad range of pH (1.67-11.80), which made it possible to evaluate the corresponding dissociation constants. 3MH and 1MH were determined together with creatinine in untreated urine samples with the limit of detection of 2.4 μM (0.4 mg LMINUS SIGN 1) and 3.0 μM (0.5 mg LMINUS SIGN 1), respectively.
Determination was fast and took ca. 12 min including the column washing. Method was employed for an analysis of urine collected from healthy individuals, and from the patients hospitalized with obesity and diabetes mellitus II.
This analysis has revealed differences between the healthy individuals and the patients pointing to a more extensive degradation of muscle proteins in the latter group.