The principle of the combined reversed-phase chromatographic electrochemical detection assay of the major serum thyroid hormones is demonstrated. The separation of 3,3',5-triiodothyronine (T(,3)), 3,3',5'-triiodothyronine (rT(,3)) and 3,5,3',5'-tetraiodothyronine (T(,4)) has been studied as a function of solvent strength, pH, temperature and ionic strength. Detection was carried out using a thin-layer electrochemical detection cell set at 1.500 V containing a new material, low temperature isotropic carbon, as a working electrode. This material was evaluated against carbon paste and using it, subnanogram detection limits were noted for the above analytes under defined conditions. Investigation of the feasibility of this analysis applied to 1.0 mL serum samples was carried out using a combined solvent extraction-cation exchange resin clean-up approach. The data indicate that under the defined conditions, elevated total T(,3) levels can be discerned and total T(,4) levels quantitated demonstrating the potential utility of this approach in screening thyrometabolic status.

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