[Determination of thyroxine enantiomers in human plasma with normal high performance liquid chromatography-chiral...

An HPLC method was developed with normal HPLC-chiral complex mobile phase for the separation and determination of D- and L-thyroxine enantiomers (D- and L-T4) in human plasma. The method includes extraction of thyroxine from plasma and separation of thyroxine enantiomers on HPLC silica column with chiral eluent containing L-proline, cupric acetate and triethylamine (TEA). The sensitivity of the method was 0.1 mg/L. The precisions of inter-day and intra-day, linearity, extraction recovery, and stability of T4 enantiomers in plasma and in deproteinized plasma were determined for the validation of the method. Baseline enantioseparation of the compounds containing D- and L-T4 was achieved. Meantime we determined the concentrations of D-T4 and L-T4 in plasma of 15 volunteers with euthyroid, hypothyroid and hyperthyroid symptoms. Liquid chromatographic method based on the described procedure was useful for the determination of D- and L-thyroxine in patient plasma and for pharmacokinetics investigation.