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Dried blood spot analysis of (+) and (−) darunavir enantiomers on immobilized amylose tris‐(3, 5‐dimethylphenylcarbamate) LC and its application to pharmacokinetics
Author(s) -
Ramisetti Nageswara Rao,
Arnipalli Manikanta Swamy,
Nimmu Narendra Varma
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.3510
Subject(s) - chromatography , chemistry , darunavir , enantiomer , dried blood spot , pharmacokinetics , whole blood , pharmacology , stereochemistry , surgery , medicine , family medicine , human immunodeficiency virus (hiv) , viral load , antiretroviral therapy
Abstract Dried blood spot analysis is an innovative novel blood sampling technique gaining interest in drug discovery and development processes owing to its inherent advantages over the conventional whole blood, plasma or serum sample collection. The present manuscript describes the development and validation of a highly sensitive and precise method of evaluation of pharmacokinetics of (+) and (−) darunavir enantiomers on rat dried blood spots. The enantiomers on rat dried blood spots were extracted into methanol and separated by LC on a Chiralpak IA column using hexane and ethanol containing 0.1% DEA (75:25, v/v) as a mobile phase at 20°C; both the enantiomers were detected at 266 nm using a photodiode array detector. The method was validated in terms of selectivity, linearity, accuracy, precision and stability as per the US Food and Drug and Administration guidelines. The hematocrit effect on extraction recovery was evaluated and the mean recoveries of (−) and (+) enantiomers of darunavir from dried blood spots were found to be 85.76 and 88.91% respectively. The intra‐ and inter‐day precision and accuracy were 3.1–8.4 and 0.8–4.8% respectively. The developed method was successfully applied to a pharmacokinetic study of (+) and (−) enantiomers of darunavir on rat dried blood spots. Copyright © 2015 John Wiley & Sons, Ltd.