الفهرس 
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Abstract
Abstract
A rapid, selective and sensitive Ultra performance liquid chromatography-mass spectrometric (UPLC-MS/MS) bio-analytical method which is based on acetonitrile precipitation technique followed by freezing to precipitated plasma proteins and lipids to minimize the matrix effect, this technique was developed and validated for the analysis of Metformin and Empagliflozin in the human plasma. The internal standards which used in the method were Metformin-d6 and Empagliflozin-d4. The separation was done by using C18 Acquity UPLC BEH column (1.7 µm, 2.1x50 mm) using isocratic elution mode that gave the excellent chromatographic separation of compounds followed by detection with tandem-mass spectrometry with the positive ionization mode (ESI+). The calibration curves which used in the method were linear over the range from 10.00 ng mL-1 to 150.00 ng/mL for Metformin and 2.0 to 250.0 ng mL-1 for Empagliflozin. The lower limit of quantitation (LLOQ) was validated at 10.00 ng mL-1 for Metformin and 2.0 ng mL-1 for Empagliflozin. The average factor of IS normalized matrix exhibited a range between 0.99 to 0.93 for Metformin relative to Metformin -d6 while it ranged between 1.04 to 0.98 for Empagliflozin relative to Empagliflozin -d4 at QCL and QCH, respectively. Results obtained indicated no significant matrix effect over the analytes ionization. The described method was successfully applied in pharmacokinetics studies and therapeutic monitoring of Metformin and Empagliflozin in plasma of 6 healthy volunteers after administrating fixed dose combination of both drug.