Quantification of N^τ -Methylhistidine and N^π-Methylhistidine in Chicken Plasma by Liquid Chromatography-Tandem Mass Spectrometry

The concentration of N^τ-methylhistidine in plasma provides an index of skeletal muscle protein breakdown. This study aimed to establish a quantitative method for measuring the concentrations of N^τ-methylhistidine and its isomer N^π-methylhistidine in chicken plasma, using liquid chromatography-tandem mass spectrometry with stable isotope dilution analysis. The acceptable linear ranges of detection were 1.56-50.00 μmol/L for N^τ-methylhistidine and 0.78-25.00 μmol/L for N^π-methylhistidine. The proposed method detected changes in the plasma levels of N^τ-methylhistidine and N^π-methylhistidine in response to fasting and re-feeding. These results suggest that the method developed in this study can be used for the simultaneous measurement of N^τ-methylhistidine and N^π-methylhistidine in chicken plasma.