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<p>A simple and accurate reverse phase high-performance liquid chromatography (RP-HPLC) tafenoquine method was developed and validated for use with bovine and equine plasma, and erythrocytes, <i>Babesia caballi</i> and <i>Theileria equi</i> culture media. After protein precipitation, samples were separated on an X-Bridge C<sub>18</sub> column using a mobile phase consisting of 11 mM ammonium acetate (pH 4.0) and acetonitrile (45:55, v/v). The extraction used acetonitrile, zinc sulfate, and hydrochloric acid. Fluorescent detection occurred with an excitation of 262&#xa0;nm and an emission of 476&#xa0;nm. The lower limit of quantification (LLOQ) was 1 ng/mL for all matrices with a sample size of 100 µL. Intra and inter-assay variability was less than 10% for all matrices and the average recovery ranged from 81% to 98%, respectively. This validated method has been used for sample evaluation in pharmacokinetic studies to evaluate tafenoquine concentrations in both equine and bovine samples as a potential candidate for controlling hemoparasites responsible for economically important diseases.</p>

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Validation of an HPLC Method for the Quantification of Tafenoquine in Bovine and Equine Plasma, Red Blood Cells, and Cell Culture Media

  • 期刊论文

  • 发表日期：

    2026年7月7日

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  • Andrew Ursini,
  • Madeline Duncan,
  • Riley Golias,
  • Julia Cutchin,
  • Reginaldo G. Bastos,
  • Nicolas F. Villarino,
  • Sherry Cox

摘要

A simple and accurate reverse phase high-performance liquid chromatography (RP-HPLC) tafenoquine method was developed and validated for use with bovine and equine plasma, and erythrocytes, Babesia caballi and Theileria equi culture media. After protein precipitation, samples were separated on an X-Bridge C18 column using a mobile phase consisting of 11 mM ammonium acetate (pH 4.0) and acetonitrile (45:55, v/v). The extraction used acetonitrile, zinc sulfate, and hydrochloric acid. Fluorescent detection occurred with an excitation of 262 nm and an emission of 476 nm. The lower limit of quantification (LLOQ) was 1 ng/mL for all matrices with a sample size of 100 µL. Intra and inter-assay variability was less than 10% for all matrices and the average recovery ranged from 81% to 98%, respectively. This validated method has been used for sample evaluation in pharmacokinetic studies to evaluate tafenoquine concentrations in both equine and bovine samples as a potential candidate for controlling hemoparasites responsible for economically important diseases.