Development and validation of RP-HPLC method with ultraviolet detection for estimation of montelukast in rabbit plasma: Application to preclinical pharmacokinetics

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Objective To develop a liquid-liquid extraction based reverse phase liquid chromatography method for estimation of montelukast in rabbit plasma. Methods Chromatographic separation was carried out using Phenomenex Luna C18 column (250 mm × 4.6 mm × 5 μm) with mobile phase composed of ammonium acetate buffer (20 Mm), pH 5.5 and acetonitrile in 20:80, v/v ratio. The analyte was monitored with UV detector at 345 nm. The developed method was validated with respect to linearity, accuracy, precision, specificity and stability. Results The peak area ratio of montelukast (MKS) to that of internal standard was used for the quantification of samples. Calibration curves were linear in the concentration range of 20-2000 ng mL-1. The LOD and LLOQ of present method were found out to be 10 ng mL-1 and 20 ng mL -1 respectively. The intra-day and inter-day %CV values for MKS were below 6.06% and 8.43%. Intra-day and inter-day accuracies were within 95.81% and 110.90%, respectively. Extraction recoveries of drug from rabbit plasma were >66.47%. Conclusion A simple, alternative, reproducible and sensitive HPLC-UV method was developed for MKS that can be used in preclinical pharmacokinetics.

Original languageEnglish
Pages (from-to)133-138
Number of pages6
JournalJournal of Young Pharmacists
Issue number4
Publication statusPublished - 12-2013


All Science Journal Classification (ASJC) codes

  • Pharmacology, Toxicology and Pharmaceutics(all)

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