HPLC Method for Analysis of Iodide and Thyroid Hormones

  Iodine is an important element in production of thyroid hormones, T3 and T4, which are essential regulators of organism's metabolism. The glands' concentration of Iodide, hormones T3 and T4, along with their precursors: 3,5-Diiodo-L-thyronine (T2), 3,5-Diiodo-L-tyrosine (DIT), and 3-Iodo-L-tyrosine (MIT) can now be studied simultaneously with this HPLC method.  In order to analyze the hydrophobic hormones along with iodide (a very polar inorganic ion) a special mixed-mode column, Primesep D, was used.  This column provides both strong hydrophobic and anion exchange properties.  The Primesep D silica-bonded ligand is comprised of a long alkyl chain and an embedded amino functional group.  The TFA was used as an ionic modifier to provide stable acidic pH and sufficient ion strength for the mobile phase.  The mobile phase composition is suitable for UV, MS, ELSD, and CAD detection.  


Column Primesep D, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN
Buffer TFA
Flow Rate 1.0 ml/min
Detection UV, 230 nm


Class of Compounds Drug,  Hydrophilic, Ionizable, Supplements
Analyzing Compounds T3, T4, 3,5-Diiodo-L-thyronine (T2), 3,5-Diiodo-L-tyrosine (DIT), and 3-Iodo-L-tyrosine (MIT)

Application Analytes:

3,5-Diiodo-L-thyronine (T2)
3,5-Diiodo-L-tyrosine dihydrate (DIT)
3-Iodo-L-tyrosine (MIT)
Levothyroxine (T4)
Liothyronine (T3)
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