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<p>A rapid, cost-effective spectrofluorometric method was developed for levothyroxine (T4) detection using fluorescein isothiocyanate (FITC) as a fluorescent probe. The method exploits FITC fluorescence quenching by T4 in Tris/phosphate buffers (pH 7.4),with linear ranges of 8.00 × 10⁻⁶ – 6.40 × 10⁻⁵ mol/L and detection limits of 9.60 × 10⁻⁶ mol/L (Tris) and 8.80 × 10⁻⁶ mol/L (phosphate). Stern-Volmer analysis confirmed dynamic quenching dominated by electrostatic interactions. The method demonstrated 102–104% recovery in spiked human serum and minimal interference from common drugs (Ciprofloxacin, Alendronate) and ions (Fe³⁺, Al³⁺). Compared to HPLC and kinetic methods, this approach offers simplicity, speed, and accessibility for routine pharmaceutical/clinical analysis.</p>

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A Rapid Fluorescence Quenching Assay for Total Levothyroxine Quantification in Pharmaceutical and Supratherapeutic Serum Samples

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月15日

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  • Enas T. Abdel-Salam,
  • Zeinab M. Anwar,
  • Ahmed Z. Ibrahim,
  • Hend M. Musatfa

摘要

A rapid, cost-effective spectrofluorometric method was developed for levothyroxine (T4) detection using fluorescein isothiocyanate (FITC) as a fluorescent probe. The method exploits FITC fluorescence quenching by T4 in Tris/phosphate buffers (pH 7.4),with linear ranges of 8.00 × 10⁻⁶ – 6.40 × 10⁻⁵ mol/L and detection limits of 9.60 × 10⁻⁶ mol/L (Tris) and 8.80 × 10⁻⁶ mol/L (phosphate). Stern-Volmer analysis confirmed dynamic quenching dominated by electrostatic interactions. The method demonstrated 102–104% recovery in spiked human serum and minimal interference from common drugs (Ciprofloxacin, Alendronate) and ions (Fe³⁺, Al³⁺). Compared to HPLC and kinetic methods, this approach offers simplicity, speed, and accessibility for routine pharmaceutical/clinical analysis.