A SIMPLE, SELECTIVE, RAPID AND RUGGED METHOD DEVELOPMENT AND VALIDATION OF TENOFOVIR AND RILPIVIRINE IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY COUPLED WITH TANDEM MASS SPECTROMETRY.
B. M. S. Kumar* and B. Rajkamal
A simple, selective, rapid and rugged liquid chromatography coupled with Tandem mass spectrometric (LC-MS/MS) method was developed and validated for the quantification of Tenofovir and Rilpivirine in human plasma. The Tenofovir and Rilpivirine were eluted within 4.00 minutes using isocratic mobile phase, the column used was Zorbax 5 Î¼, C18, 100Ã—4.60 mm having a mobile phase of 5 mM Ammonium acetate, 20.0% and Acetonitrile, 80% (20:80% v/v). The flow rate was 0.70 ml/min at a column temperature of 40 Â± 5ÂºC. Analysis was performed using multiple reaction monitoring (MRM) and ionization was carried out using electrospray positive ionization mode. The retention time for Tenofovir and Rilpivirin are 0.85 min and 2.80 min respectively. The method was validated for linearity, precision, accuracy, specificity, sensitivity, matrix effect, dilution integrity, ruggedness, injection reproducibility and stability. Calibration curve range for Tenofovir and Rilpivirine were 5.000 â€“ 600.000 and 1.000 -203.000 ng/mL respectively. The calibration curves were linear during the course of validation, with correlation coefficients â‰¥0.9988 and > 0.9992 for Tenofovir and Rilpivirine respectively. The precision and mean accuracy were within the acceptable limits.
Keywords: Tenofovir; Rilpivirine; LC/MS/MS; Validation.
[Full Text Article]