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- Title
Development and Validation of a lc-ms/ms Method for the Determination of Raltegravir in Sprague Dawley Rat Serum and Its Application to Pharmacokinetic Study.
- Authors
Raju, Vijaya A. D.; Nemala, Appala Raju
- Abstract
A novel bio-analytical method was developed and validated for the quantitative determination of raltegravir in rat serum by using the liquid-liquid extraction chromatography and tandem mass spectrometric detection (HPLC-MS/MS). Separation of raltegravir from the endogenous substances is achieved after liquid-liquid extraction by using HPLC-MS/MS system. Raltegravir was eluted in isocratic mode with acetonitrile, methanol and 0.1% acetic acid in water (40:30:30) at a flow-rate of 0.5 mL/min on Waters, Exterra C18, 50*4.6 mm, 5µm particle size column. Didanosine was used as the internal standard. The liquid-liquid extraction recovery was found 70% indicates good recovery. The validation results demonstrated that the present method was found to be precise and accurate. The stability tests indicated that the raltegravir in rat serum is stable for three freeze-thaw cycles at both -20 ºC and -70 ºC, 18-h ambient storage, 15-day frozen storage at both -20 ºC and -70 ºC. The results also showed no significant matrix effect (<6.2%). The present method was found to be sensitive and selective at very low levels of linearity range 1-1000 ng/mL, based on a sample volume of 50 µL, with a linear correlation coefficient of ≥ 0.99. The validated method has been successfully applied to support a preclinical pharmacokinetic study.
- Subjects
RALTEGRAVIR; PHARMACOKINETICS; ACETONITRILE; METHANOL; PHYSIOLOGICAL effects of acetic acid
- Publication
American Journal of Biomedical Sciences, 2013, Vol 5, Issue 3, p197
- ISSN
1937-9080
- Publication type
Article
- DOI
10.5099/aj130300197