Development and validation of a simple and isocratic reversed-phase HPLC method for the determination of rilpivirine from tablets, nanoparticles and HeLa cell lysates

Abhijit A. Date, Annemarie Shibata, Patrick Bruck, Christopher J. Destache

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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 C18 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 (r2 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.

Original languageEnglish
Pages (from-to)709-715
Number of pages7
JournalBiomedical Chromatography
Volume29
Issue number5
DOIs
StatePublished - May 1 2015

Fingerprint

Rilpivirine
HeLa Cells
Nanoparticles
Tablets
High Pressure Liquid Chromatography
Dosage Forms
Limit of Detection
Flow rate

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Drug Discovery
  • Pharmacology

Cite this

@article{79293b5994ff43cdbd3c84afe725e6cc,
title = "Development and validation of a simple and isocratic reversed-phase HPLC method for the determination of rilpivirine from tablets, nanoparticles and HeLa cell lysates",
abstract = "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 C18 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 (r2 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.",
author = "Date, {Abhijit A.} and Annemarie Shibata and Patrick Bruck and Destache, {Christopher J.}",
year = "2015",
month = "5",
day = "1",
doi = "10.1002/bmc.3346",
language = "English",
volume = "29",
pages = "709--715",
journal = "Biomedical Chromatography",
issn = "0269-3879",
publisher = "John Wiley and Sons Ltd",
number = "5",

}

TY - JOUR

T1 - Development and validation of a simple and isocratic reversed-phase HPLC method for the determination of rilpivirine from tablets, nanoparticles and HeLa cell lysates

AU - Date, Abhijit A.

AU - Shibata, Annemarie

AU - Bruck, Patrick

AU - Destache, Christopher J.

PY - 2015/5/1

Y1 - 2015/5/1

N2 - 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 C18 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 (r2 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.

AB - 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 C18 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 (r2 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.

UR - http://www.scopus.com/inward/record.url?scp=84927731911&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84927731911&partnerID=8YFLogxK

U2 - 10.1002/bmc.3346

DO - 10.1002/bmc.3346

M3 - Article

C2 - 25298145

AN - SCOPUS:84927731911

VL - 29

SP - 709

EP - 715

JO - Biomedical Chromatography

JF - Biomedical Chromatography

SN - 0269-3879

IS - 5

ER -