Development and Validation of a Dual Column HPLC Method for Determination of Albendazole and Its Metabolites in Rat Plasma

To develop a quick, simple and reproducible dual column high performance liquid chromatography (HPLC) method to determine the albendazole and its metabolites in rat plasma. Albendazole (ABZ), albendazole sulfoxide (ABZSO) and albendazole sulfone (ABZSO2) were analyzed in rat plasma by high performance liquid chromatography using UV-detector. Preparation of plasma samples was carried out by protein precipitation using 8.25% perchloric acid. This method involves two different mobile phases with two different columns and different wavelengths. Estimation of Albendazole was done using Enable C18 column (250 mm x 4.6 mm, 5 mu m: SpinCo Biotech Pvt. Ltd.), mobile phase acetonitrile: water in the ratio 60: 40, wavelength 225nm and Praziquantel as an internal standard (IS). The retention time for Albendazole and Praziquantel was 3.7 and 6.4 minutes respectively. But the estimation of Albendazole Sulfoxide, Albendazole Sulfone were done by using Phenomenex C18 Luna column (250 mm x 4.6 mm, 5 mu m: USA), mobile phase acetonitrile: methanol: phosphate buffer (20mM) in the ratio 20: 25: 55. The pH was adjusted to 6.9 using 0.1N NaOH solution, wavelength 290nm and oxfendazole an internal standard (IS). The retention time for Albendazole Sulfoxide, Albendazole Sulfone, and Oxfendazole was 5.5, 7.0 and 8.2 minutes respectively. Both the methods were validated over the range from 0.005-5 mu g/mL for Albendazole, 0.0580 mu g/mL for Albendazole Sulfoxide and Albendazole Sulfone. Both the method showed % RSD and % DEV lower than 15% for all the analytes. The limit of quantitation was 0.005 mu g/mL for Albendazole whereas 0.05 mu g/mL for Albendazole Sulfoxide and Albendazole Sulfone. Metabolites of albendazole were analyzed in rat plasma samples using a single dose of Albendazole 50mg/kg was determined application of this method was also used to found the pharmacokinetic studies.

Automated summary

A quick, simple and reproducible dual column high performance liquid chromatography (HPLC) method was developed to determine albendazole and its metabolites in rat plasma. This method involves two different mobile phases, two different columns and different wavelengths, allowing accurate quantification of the target compounds.