Development and validation of a sensitive LC-MS/MS method with electrospray ionization for quantitation of rhein in human plasma: Application to a pharmacokinetic study

Buddhadev Layek, T. Santosh Kumar, Ravi Kumar Trivedi, Ramesh Mullangi, Nuggehally R. Srinivas

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

A highly sensitive and specific LC-MS/MS method has been developed and validated for the estimation of rhein with 100 μL human plasma using celecoxib as an internal standard (IS). The API-4000 Q-Trap LC-MS/MS was operated under multiple reaction-monitoring mode using the electrospray ionization technique. The assay procedure involved extraction of rhein and IS from human plasma with acetonitrile, which yielded consistent recoveries of 36.01 and 65.85% for rhein and IS, respectively. The total chromatographic run time was 5.0 min and the elution of rhein and IS occurred at ∼1.60 and 3.96 min, respectively. The resolution of peaks was achieved with 0.01 M ammonium acetate (pH 6.0):acetonitrile:methanol (30:58:12, v/v) on an Inertsil ODS-3 column. The method was proved to be accurate and precise at a linearity range of 0.005-5.00 μg/mL with a correlation coefficient (r) of ≥0.995. The lower limit of quantitation was 0.005 μg/mL. The intra- and inter-day precision and accuracy values were found to be within the assay variability limits as per the FDA guidelines. Rhein was found to be stable in the battery of stability studies. The application of the assay to pre-clinical pharmacokinetic studies confirmed the utility of the assay to derive pharmacokinetic parameters.

Original languageEnglish (US)
Pages (from-to)616-624
Number of pages9
JournalBiomedical Chromatography
Volume22
Issue number6
DOIs
StatePublished - Jun 1 2008

Keywords

  • Human plasma
  • LC-MS/MS
  • Pharmacokinetics
  • Rhein
  • Validation

Fingerprint

Dive into the research topics of 'Development and validation of a sensitive LC-MS/MS method with electrospray ionization for quantitation of rhein in human plasma: Application to a pharmacokinetic study'. Together they form a unique fingerprint.

Cite this