Determination of Doxycycline in Human Plasma by Liquid Chromatography-Mass Spectrometry after Liquid-Liquid Extraction and its Application in Human Pharmacokinetics Studies
Muralidharan Selvadurai, Subramania Nainar Meyyanathan*, Sekar Rajan, Govindarajan Padmanaban and Bhojraj Suresh
Department of Pharmaceutical Analysis, J.S.S.College of pharmacy, Ootacamund 643001, Tamilnadu, India
- *Corresponding Author:
- Dr. Subramania Nainar Meyyanathan Ph.D
Professor and Head
Department of pharmaceutical Analysis
J.S.S.College of pharmacy
Ootacamund-643001, Tamilnadu, India
E-mail: [email protected]
Received Date: July 15, 2010; Accepted Date: September 16, 2010; Published Date: September 16, 2010
Citation: Selvadurai M, Meyyanathan SN, Rajan S, Padmanaban G, Suresh B (2010) Determination of Doxycycline in Human Plasma by Liquid Chromatography- Mass Spectrometry after Liquid-Liquid Extraction and its Application in Human Pharmacokinetics Studies. J Bioequiv Availab 2: 093-097. doi: 10.4172/jbb.1000038
Copyright: © 2010 Selvadurai M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
A sensitive and reproducible liquid chromatography–mass spectrometric (LC-MS) method has been developed and validated for the quantifi cation of doxycycline in human plasma, after liquid-liquid extraction (LLE). Best chromatographic resolution was achieved on a reverse-phase Phenomenex C18 column using the mobile phase acetonitrile–5mM ammonium acetate (80:20) in isocratic elution with a total run time of 3.5 min. The analyte, doxycycline was detected by using an electrospray ionization mass spectrometer in the selected ion monitoring (SIM) mode. Linear plot was obtained in the concentration range of 0.5–5.0 µg/ml (r2 = 0.998). Lower limit of quantifi cation (LLOQ) was found to be 100 ng/ml in 500µl of plasma. Average recovery of the analyte was found to range from 96.08 to 104.60% in plasma at the concentrations of 1.0, 3.0 and 5.0 µg/ml. Intra- and inter-day relative standard deviation was found to be 0.13-0.74 and 0.11-0.34 respectively. The present method was successfully applied in the pharmacokinetic study of doxycycline in human plasma.