Development of a validated HPLC method for the determination of iodotyrosines and iodothyronines in pharmaceuticals and biological samples using solid phase extraction

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-01, Vol.814 (1), p.163-172
• Main Authors: Gika, Helen G., Samanidou, Victoria F., Papadoyannis, Ioannis N.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Iodothyronines; Iodotyrosines; Medical sciences; Monoiodotyrosine - analysis; Pharmaceutical Preparations - chemistry; Pharmaceuticals; Pharmacology. Drug treatments; Reference Standards; Serum; SPE biological samples; Thyroid gland hormones; Thyronines - analysis; Tissue; Urine
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2004.10.025

Identification, separation and quantitation of iodoaminoacids, is essential for the biological research and the clinical diagnosis of thyroid gland disease. Under this aspect a reversed-phase high-performance liquid chromatographic method was developed for the determination of thyroid gland hormones and some of their primary metabolites, 3,3′,5,5′-tetra-iodo- l-thyronine ( l-thyroxine), 3,3′,5-tri-iodo- l-thyronine, 3,5-di-iodo- l-thyronine, l-thyronine, 3,5-di-iodo- l-tyrosine, 3-iodo- l-tyrosine and l-tyrosine. Analysis was performed on an Inertsil C 18 column with photodiode-array detection, using a 25 min gradient scale program of a binary mobile phase consisted of 0.1% aqueous solution of trifluoroacetic acid at pH 3 as solvent A and acetonitrile as solvent B, at a flow rate of 1 mL/min. Quantitation was performed using were obtained using theophylline as internal standard. The method was applied to commercial pharmaceuticals and biological samples (serum, urine and tissue). Drug-free urine and serum samples were spiked with known concentrations of the analytes standards and pretreated by solid phase extraction to remove matrix interferences. C 18 cartridges were used, yielding recoveries ranging from 87.1% to 107.6% for serum samples and from 92.1% to 98.7% for urine samples. With regard to total-T 4 concentrations in serum samples, results are cross-validated with RIA and found to agree well.