Bio-analytical method development of antimalarial drug in human plasma
Validation for quantitative estimation by using LC-MSMS
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In the present work new, simple reversed-phase LCMS/MS in positive ion mode method was developed and validated for the determination of an anti-malarial in blood plasma. Drug was used as an internal standard. The chromatographic separation was achieved with Zorbax SB-C18, 5.0µm (150*4.6mm). The separation of Drug H was achieved by gradient elution using organic mixture (Acetonitrile: Methanol::80:20 v/v), buffer solution (10mM Ammonium Formate) and DMSO:: 50:50:0.2 v/v/v. The flow rate is 1.000 ml/min, Injection Volume of 10µl with a run time of 3.0 minutes. The calibration curve of standard...
In the present work new, simple reversed-phase LCMS/MS in positive ion mode method was developed and validated for the determination of an anti-malarial in blood plasma. Drug was used as an internal standard. The chromatographic separation was achieved with Zorbax SB-C18, 5.0µm (150*4.6mm). The separation of Drug H was achieved by gradient elution using organic mixture (Acetonitrile: Methanol::80:20 v/v), buffer solution (10mM Ammonium Formate) and DMSO:: 50:50:0.2 v/v/v. The flow rate is 1.000 ml/min, Injection Volume of 10µl with a run time of 3.0 minutes. The calibration curve of standard Drug H was linear in range 0.1-20.0 ¿g/ml. 5 precision and accuracy batches were run; the coefficient of correlation (r2) was ¿ 0.99. The accuracy of the LLOQ in the standard CC was within ± 20% of the nominal concentration and ± 15% of the nominal concentration for other standards and 75% non- zero calibration standards met the acceptable criteria which indicated the method was precise and accurate. Method validation results have proved the method to be selective, sensitive, precise, accurate and robustness.
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