Title:Development and Validation of RP-HPLC Method for Roflumilast Nano Particles in Rat Plasma for Pharmacokinetic Study Analysis
Volume: 20
Issue: 9
Author(s): Yasir Mehmood*Hira Shahid
Affiliation:
- Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Government College University Faisalabad,
38000, Faisalabad, Pakistan
Keywords:
ICH guidelines, RP-HPLC, determination, roflumilast, quantification, plasma.
Abstract:
Aims: The aim of this study is to develop a sensitive, specific, rapid, and precise reverse-
phase high-performance liquid chromatography (RP-HPLC) method and validate it to determine
the Roflumilast in rat plasma.
Background: This study was aimed at developing a simple HPLC method for the detection of Roflumilast
in rat plasma after ingestion of nanoparticles into the rat. A bioanalytical method was developed
and validated.
Methodology: The drug sample (Roflumilast) was eluted isocratically using a mobile phase of Ammonium
acetate buffer (pH = 7.0), acetonitrile, and methanol (20:40:40, v/v/v) made up the mobile
phase on a Phenomenex C-18 (150mm x 4.68mm pore size 5-micron analytical column. The
drug was quantitatively determined at a UV wavelength of 254 nm using a standard calibration
curve spanning the range of 1.5, 3.0, 4.5, 6.0, 7.5μg/mL-1.
Results: The results showed that the limits of quantitation (LOQ) and detection (LOD) were 0.18
μg/ml and 0.54μg/ml, respectively. For both the intra-day and inter-day analysis, relative standard
deviation (% RSD) was less than 2%, and the drug’s percentage accuracy was between 96% and
98%.
Conclusion: As per the established protocols, the developed method was quantitatively assessed
for its intended use in terms of specificity, linearity, accuracy, precision, robustness, and sensitivity
analysis. Roflumilast-loaded nanoparticles were effectively used to determine the drug entrapment
efficiency using the currently validated RP-HPLC method. Besides, highly efficient drug extraction
from plasma was attained, and mobile phase composition was suitable to the pharmacokinetic
study of Roflumilast-loaded nanoparticles in rats.