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Determination of Doxycycline in Human Plasma by Liquid Chromatography-Mass Spectrometry after Liquid-Liquid Extraction and its Application in Human Pharmacokinetics Studies

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Author(s): Muralidharan Selvadurai | Subramania Nainar Meyyanathan | Sekar Rajan | Govindarajan Padmanaban | Bhojraj Suresh

Journal: Journal of Bioequivalence & Bioavailability
ISSN 0975-0851

Volume: 2;
Issue: 4;
Start page: 093;
Date: 2010;
Original page

Keywords: Doxycycline | LC-MS | Validation | Pharmacokinetic studies

ABSTRACT
A sensitive and reproducible liquid chromatography–mass spectrometric (LC-MS) method has been developed andvalidated for the quantifi cation of doxycycline in human plasma, after liquid-liquid extraction (LLE). Best chromatographicresolution was achieved on a reverse-phase Phenomenex C18 column using the mobile phase acetonitrile–5mMammonium acetate (80:20) in isocratic elution with a total run time of 3.5 min. The analyte, doxycycline was detectedby using an electrospray ionization mass spectrometer in the selected ion monitoring (SIM) mode. Linear plot wasobtained in the concentration range of 0.5–5.0 μg/ml (r2 = 0.998). Lower limit of quantifi cation (LLOQ) was found to be100 ng/ml in 500μl of plasma. Average recovery of the analyte was found to range from 96.08 to 104.60% in plasma atthe concentrations of 1.0, 3.0 and 5.0 μg/ml. Intra- and inter-day relative standard deviation was found to be 0.13-0.74and 0.11-0.34 respectively. The present method was successfully applied in the pharmacokinetic study of doxycyclinein human plasma.
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