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

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.

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