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

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 sul...

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Main Authors: E. Milena Vázquez, Beatriz Romero, Ana M. Sahagún, Cristina López, Raúl de la Puente, José M. Rodríguez, Nélida Fernández, M. José Diez, Raquel Díez
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/9/2039
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author E. Milena Vázquez
Beatriz Romero
Ana M. Sahagún
Cristina López
Raúl de la Puente
José M. Rodríguez
Nélida Fernández
M. José Diez
Raquel Díez
author_facet E. Milena Vázquez
Beatriz Romero
Ana M. Sahagún
Cristina López
Raúl de la Puente
José M. Rodríguez
Nélida Fernández
M. José Diez
Raquel Díez
author_sort E. Milena Vázquez
collection DOAJ
description 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<sup>®</sup> 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).
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spelling doaj-art-8cd1cc9871b7456caec966244d0c3ceb2025-08-20T01:49:14ZengMDPI AGMolecules1420-30492025-05-01309203910.3390/molecules30092039Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation StudyE. Milena Vázquez0Beatriz Romero1Ana M. Sahagún2Cristina López3Raúl de la Puente4José M. Rodríguez5Nélida Fernández6M. José Diez7Raquel Díez8Pharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainPharmacology, Department of Biomedical Sciences, Veterinary Faculty, Institute of Biomedicine (IBIOMED), University of Leon, 24071 Leon, SpainAlbendazole 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<sup>®</sup> 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).https://www.mdpi.com/1420-3049/30/9/2039albendazolealbendazole sulfoxidealbendazole sulfoneEMA guidelineHPLCvalidation
spellingShingle E. Milena Vázquez
Beatriz Romero
Ana M. Sahagún
Cristina López
Raúl de la Puente
José M. Rodríguez
Nélida Fernández
M. José Diez
Raquel Díez
Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
Molecules
albendazole
albendazole sulfoxide
albendazole sulfone
EMA guideline
HPLC
validation
title Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
title_full Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
title_fullStr Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
title_full_unstemmed Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
title_short Analytical Method for the Simultaneous Determination of Albendazole and Metabolites Using HPLC-PDA: A Validation Study
title_sort analytical method for the simultaneous determination of albendazole and metabolites using hplc pda a validation study
topic albendazole
albendazole sulfoxide
albendazole sulfone
EMA guideline
HPLC
validation
url https://www.mdpi.com/1420-3049/30/9/2039
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