High Performance Liquid Chromatography Fluorescence Method for the Determination of Seleno-Amino Acids in Ovine Blood Plasma

This study was performed to validate an effective high performance liquid chromatography-fluorescence detection method for the separation of seleno-amino acids and sulfur amino acids in biological samples using pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Standard calibration and standard addition procedures were developed, but the quantification of seleno-amino acids was carried out by a second method. The coefficient of variation for precision was < 15% and accuracies were 96 to 98%. Recoveries in the extraction method were between 63 and 99%. This method was applied to the contribution of seleno-amino acids containing methylselenocysteine and selenomethionine in plasma tissue of lambs during infusion study. The method demonstrated good selectivity, sensitivity, precision, and accuracy.