Bio-analytical method development of antimalarial drug in human plasma. Validation for quantitative estimation by using LC-MSMS

      Аннотация: 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.

Ключевые слова: Bio-analytical method development, LC-MSMS, ANTI-MALARIAL DRUG, human plasma, validation