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Rational design for variability minimization in bioanalytical method validation: illustration with LC‐MS/MS assay method for terbinafine estimation in human plasma
Author(s) -
Gurule Sanjay,
Khuroo Arshad,
Monif Tausif,
Goswami Dipanjan,
Saha Arabinda
Publication year - 2010
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.1423
Subject(s) - bioanalysis , chromatography , terbinafine , chemistry , analyte , extraction (chemistry) , solid phase extraction , detection limit , human plasma , antifungal , medicine , itraconazole , dermatology
Abstract Terbinafine, a widely used antifungal drug, is a challenging molecule for quantitative bioanalysis due to certain factors contributing assay variability. Despite previous attempts at human plasma determination of terbinafine, exhaustive stability of the drug or an internal standard was lacking. Internal standard stability with negligible variation throughout the analysis is an indicator of a reliable bioanalytical method as the majority of LC–MS/MS assays are based on analyte/IS response ratios for quantitation. A newly developed high‐throughput simple LC‐MS/MS method is described for human plasma determination of terbinafine using naftifine internal standard and eluting all compounds within 2 min. A solid‐phase extraction of terbinafine achieving mean recovery of 84.3% (CV < 4%) without compromising sensitivity (limit of quantitation 5.11 ng/mL) or linearity (5.11–3014.19 ng/mL) is delineated in this paper. A heated nebulizer in positive multiple reaction monitoring mode was employed with transitions m/z 292.2 →141.1 and 288.2 →117.0 for terbinafine and naftifine, respectively, resulting in excellent chromatographic separation on a Hypurity Advance (50 × 4.6 mm, 5 µm) column. The developed method was successfully applied to clinical samples and for the first time demonstrated marked improved extraction efficiency and reliable long‐term plasma stability results without any internal standard response variation during the entire course of study. Copyright © 2010 John Wiley & Sons, Ltd.