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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
Author(s) -
Layek Buddhadev,
Kumar T. Santosh,
Trivedi Ravi Kumar,
Mullangi Ramesh,
Srinivas Nuggehally R.
Publication year - 2008
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.977
Subject(s) - chemistry , chromatography , electrospray ionization , ammonium acetate , pharmacokinetics , detection limit , extraction (chemistry) , human plasma , selected reaction monitoring , acetonitrile , ethyl acetate , mass spectrometry , tandem mass spectrometry , high performance liquid chromatography , medicine
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. Copyright © 2008 John Wiley & Sons, Ltd.