DETERMINATION OF NIFEDIPINE IN RAT PLASMA USING HPLC-UV DETECTOR: A SIMPLE METHOD FOR PHARMACOKINETICS AND ORAL BIOAVAILABILITY STUDIES
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2016, Vol 8, Issue 8
Abstract
Objective: To develop and validate a high-performance liquid chromatographic method (HPLC) for the determination of nifedipine (NFD) concentration in rat plasma.Methods: 1.5 mol of sodium hydroxide solution was added to each plasma sample, followed by the addition of an extraction solvent based on n-hexane and dichloromethane (70: 30, v/v). The organic layer was transferred and evaporated to dryness under nitrogen flow. The residue was reconstituted with 0.5 mol of acetic acid, followed by the addition of n-hexane. After centrifuging the mixture, the supernatant organic layer of n-hexane was discarded, and the aqueous solution was injected onto the HPLC using A Phenomenex Luna-C18 reversed phase analytical column (250 x 4.6 mm, 5 µm). The mobile phase consisted of 0.01 mol aqueous ammonium formate: methanol: acetonitrile (55: 43: 2, v/v) with pH adjusted to 4.9 using formic acid. The flow rate was 0.8 ml/min; UV detector set at 235 nm and the samples were quantified using the peak area.Results: A well-resolved NFD peak was achieved free of interference from endogenous compounds in rat plasma. Recovery of NFD was more than 93 % over concentrations ranged from 5.00 to 200 ng/ml. The limit of quantification (LOQ) of this assay was 6 ng/ml and, intra-and inter-day coefficient of variation (CV) were 5.75 % and 7.93 %, respectively. NFD was found to be stable in rat plasma after being stored at -30 °C over 90 d.Conclusion: The stability, sensitivity, specificity and reproducibility of this method make it suitable for the determination of NFD plasma concentration in pharmacokinetics and oral bioavailability studies.
Authors and Affiliations
Mosab Arafat, Zahaa Ahmed, Momir Mikov
Keywords
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