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Development and validation of a simple and isocratic reversed‐phase HPLC method for the determination of rilpivirine from tablets, nanoparticles and HeLa cell lysates
Author(s) -
Date Abhijit A.,
Shibata Annemarie,
Bruck Patrick,
Destache Christopher J.
Publication year - 2015
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3346
Subject(s) - chemistry , rilpivirine , hela , chromatography , detection limit , acetonitrile , high performance liquid chromatography , dosage form , nanoparticle , potassium , quantitative analysis (chemistry) , analytical chemistry (journal) , materials science , nanotechnology , cell , human immunodeficiency virus (hiv) , medicine , biochemistry , organic chemistry , family medicine , viral load , antiretroviral therapy
In the present investigation, a simple and isocratic HPLC‐UV method was developed and validated for determination of rilpivirine (RPV) from dosage forms (tablets and nanoparticles) and biological matrices like HeLa cell lysates. The separation and analysis of RPV was carried out under isocratic conditions using (a) a Gemini reversed‐phase C 18 column (5 µm; 4.6 × 150 mm) maintained at 35°C, (b) a mobile phase consisting of a mixture of acetonitrile and 25 m m potassium dihydrogen phosphate (in the ratio 50:50 v/v) at a flow rate of 0.6 mL/min and (c) atazanavir as an internal standard. The total run time was 17 min and the analysis of RPV and internal standard was carried out at 290 nm. The method was found to be linear ( r 2 value > 0.998), specific, accurate and precise over the concentration range of 0.025–2 µg/mL. The lower limit of quantification was 0.025 µg/mL, the limit of detection was 0.008 µg/mL and the recovery of RPV was >90%. The stability of the RPV analytical method was confirmed at various conditions such as room temperature (24 h), −20°C (7 days), three freeze−thaw cycles and storage in an autosampler (4°C for 48 h). The method was successfully applied for the determination of RPV from conventional dosage forms like tablets, from polymeric nanoparticles and from biological matrices like HeLa cell lysates. Copyright © 2014 John Wiley & Sons, Ltd.