Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study

Molecules. 2025 May 3;30(9):2039. doi: 10.3390/molecules30092039.

Abstract

Albendazole is an anthelmintic drug commonly used in animals and humans against nematodes. A sensitive, accurate, precise, and time-saving high-performance liquid chromatography (HPLC) method for the simultaneous determination of albendazole and metabolites (albendazole sulfoxide and albendazole sulfone) in cattle plasma has been developed and validated. A solid-phase extraction (SPE) was carried out. Separation was performed with an XBridge® C18 column (4.6 mm × 250 mm, 5 µm) with gradient elution of acetonitrile:ammonium acetate buffer 0.025 M with pH adjusted to 6.6. The flow rate was 1.2 mL/min, and the PDA detector was set at 292 nm. Calibration curves were linear in the range from 0.025 to 2.0 µg/mL for the three compounds evaluated, with correlation coefficients ≥ 0.99. For the lower limit of quantification (LLOQ), within- and between-run precision and accuracy were satisfactory, with coefficients of variation (CV) ≤ 15.1% and deviations ≤ 117.7%, respectively. The method fulfilled all validation criteria established by the European Medicines Agency guideline (EMA/CHMP/ICH/172948/2019).

Keywords: EMA guideline; HPLC; albendazole; albendazole sulfone; albendazole sulfoxide; validation.

Publication types

  • Validation Study

MeSH terms

  • Albendazole* / analogs & derivatives
  • Albendazole* / blood
  • Albendazole* / metabolism
  • Animals
  • Anthelmintics* / blood
  • Cattle
  • Chromatography, High Pressure Liquid / methods
  • Limit of Detection
  • Reproducibility of Results
  • Solid Phase Extraction

Substances

  • Albendazole
  • albendazole sulfoxide
  • Anthelmintics
  • albendazole sulfone