Method for the quantitation of iodothyronines in body tissues and fluids using high-performance liquid chromatography
| Autor: | Joseph Berdecia-Rodriguez, Susan P. Porterfield, Chester E. Hendrich, Vernon T. Wiedmeier |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Brain Chemistry
Thyroid Hormones Chromatography Resolution (mass spectrometry) Phloretin Sodium Reproducibility of Results chemistry.chemical_element General Chemistry High-performance liquid chromatography Chloride Fluorescence Rats chemistry.chemical_compound Spectrometry Fluorescence Liver chemistry medicine Animals Amine gas treating Iodotyrosine Chromatography High Pressure Liquid medicine.drug |
| Zdroj: | Journal of Chromatography B: Biomedical Sciences and Applications. 577:19-24 |
| ISSN: | 0378-4347 |
| DOI: | 10.1016/0378-4347(92)80594-g |
| Popis: | The separation and quantitation of iodotyrosines and iodothyronines [3-monoiodo-L-tyrosine, 3,5-diiodo-L-tyrosine, 3,5-, 3,3' and 3',5'-diiodo-L-tyronines, 3,5,3'-triiodo-L-thyronine (T3), reverse 3,3',5'-triiodo-L-thyronine and 3,3',5,5'-tetraiodo-L-thyronine (T4)] from animal tissues (brain, liver and serum) by a new high-performance liquid chromatographic (HPLC) method is described. Rats were infused with iso-osmotic sodium chloride containing 100 microM phloretin to block deiodination. The tissues were extracted using differential pH values to separate other amines from the amine containing iodothyroid hormones. Aliquots of tissue extracts (25-100 microliters) were reacted overnight with 5-dimethylaminonaphthalene-1-sulfonyl chloride and their iodotyrosine and iodothyronine content determined by HPLC utilizing fluorimetric detection. Resolution of the individual compound peaks was achieved by gradient elution with a 3.0 mM H3PO4 buffer. Greater sensitivity has been achieved (less than 1.0 pmol/g) utilizing fluorescence rather than ultraviolet absorbance for the quantitation of these iodinated compounds. The method is superior also to other methods in that recoveries, based on those of 125I-labelled T4 and T3, were 89-97%. |
| Databáze: | OpenAIRE |
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