HIGH THROUGHPUT LCMS-MS METHOD FOR THE QUANTITATION OF SAXAGLIPTIN IN RAT PLASMA BY PROTEIN PRECIPITATION USING 96 WELL PLATE FORMAT
Tangudu Nagabhusana Rao*, Guntuku Girija Sankar, Lade Jyothi Rani
ABSTRACT
High-throughput Liquid chromatography–mass spectrometry method has been developed and validated for the quantification of Saxagliptin in rat plasma using Sitagliptin as internal standard (ISTD). Following protein precipitation in 96 well plate format, the analytes and ISTD were run on ACE C18 4.6 x75 mm (5.0 μm) using an isocratic mobile phase consisting of 2 mM Ammonium Formate with 0.1% Formic Acid and Acetonitrile (50:50 v/v). The precursor and product ions of the drugs were monitored on a triple quadrupole instrument operated in the positive ionization mode. The method was validated over a concentration range of 0.997 to 249.259 ng/mL with mean recovery of 104.74%. The inter batch precision (%CV) across three validation runs was ≤ 10.5%. The Inter batch accuracy determined at four QC levels (LLOQ, LQC, MQC and HQC) was between 101.3 – 103.6%. According to the validated results, the proposed method was found to be specific, accurate, precise and high throughput method. This method could be used for the estimation of Saxagliptin in rat plasma and can be applied for the routine analysis.
Keywords: Saxagliptin, Sitagliptin, Protein Precipitation and 96 well plates.
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