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JYP · Lens · 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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Article · Volume 5, Issue 4 · pp. 133–138

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

Om Prakash Ranjan1, Usha Y. Nayak1, Meka Sreenivasa Reddy1, Swapnil J. Dengale2, Prashant B. Musmade2, Nayanabhirama Udupa3

  1. 1Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, INDIA.
  2. 2Department of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, INDIA.
  3. 3Department of Pharmaceutical Management, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, INDIA.

Abstract

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 (250mm X 4.6mm X 5 mm) with mobile phase composed of ammoniumacetate buffer (20Mm), pH 5.5 and acetonitrile in 20:80, v/v ratio. The analytewasmonitoredwithUV detector at 345nm. The developedmethod 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 20e2000 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.

Keywords: Montelukast; Rabbit plasma; HPLC; UV detection; Pharmacokinetics

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