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Simultaneous determination of raltegravir and raltegravir glucuronide in human plasma by liquid chromatography–tandem mass spectrometric method
Author(s) -
Wang LingZhi,
Lee Lawrence SoonU,
Thuya WinLwin,
Soon Gaik Hong,
Kong LiRen,
Nye PeiLing,
Deoon Lee Edmund Jon,
Flexner Charles,
Goh BoonCher
Publication year - 2011
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1874
Subject(s) - raltegravir , chemistry , chromatography , glucuronide , tandem mass spectrometry , mass spectrometry , liquid chromatography–mass spectrometry , selected reaction monitoring , pharmacokinetics , metabolite , pharmacology , biochemistry , medicine , family medicine , human immunodeficiency virus (hiv) , viral load , antiretroviral therapy
Raltegravir is a highly efficacious inhibitor of HIV integrase. Large pharmacokinetic variability has been reported in clinical trials and this could be due to glucuronidation of raltegravir, the only reported metabolism pathway. In order to precisely evaluate and monitor the raltegravir and raltegravir glucuronide simultaneously, a novel, sensitive and robust liquid chromatography‐tandem mass spectrometric method was developed and validated for simultaneous determination of raltegravir and raltegravir glucuronide in human plasma. A simple protein precipitation with acetonitrile was utilized for plasma sample preparation prior to analysis. Baseline chromatographic separation was achieved on a ZORBAX Eclipse XDB‐C8 using gradient elution mode. The run time was 9 min at a constant flow rate of 0.4 ml/min. The mass spectrometer was operated under a positive electrospray ionization condition. Excellent linearity ( r 2 ≥ 0.9997) was achieved for raltegravir and raltegravir glucuronide in the range of 2–2000 nmol/l. The average recovery of raltegravir and raltegravir glucuronide was 105.8% and 102.2%, respectively. The precision (coefficient of variation) was 1.6–6.6% for raltegravir and 2.1–6.9 for raltegravir glucuronide, respectively. The accuracy was 98.6–106.1% for raltegravir and 96.3–100.3% for raltegravir glucuronide. The plasma samples were tested to be stable after nine freeze–thaw cycles and exposure to room temperature for 24 h. This well‐validated assay was applied for the quantification of raltegravir and raltegravir glucuronide in plasma samples within 24 h after a single oral dose of 400 mg raltegravir in six healthy subjects. Copyright © 2011 John Wiley & Sons, Ltd.