Method Development and Validation For Determining Stability of Omadacycline In Biological Matrices By Liquid Chromatography–Mass Spectrometry

The validated protein precipitation method was applied for the estimation of Omadacycline (OM) in human plasma with Omadacycline-D9 (OMD9) as an internal standard (ISTD) by using HPLC-ESI-MS/MS. Zorbax Eclipse Plus C18, 2.1 x 50 mm, 3.5 μm, was selected as the analytical column. The column temperature was set at 45°C. Mobile phase composition was 0.1% formic acid: methanol (80:20 v/v). Source flow rate of 300 μL/min without a split. An injection volume of 10 μL. Omadacycline and Omadacycline-D9 mesylate were eluted at 1.2 ± 0.2 min, with a total run time of 3.0 min for each sample. The mass transitions of Omadacycline and Omadacycline-D9 obtained were m/z 557.6 ® 456.6 and 566.7 ® 456.6, respectively. The standard curve shows a correlation coefficient (r2) greater than 0.9983 with a range of 5.00 to 12000.00 pg/ml using the linear regression model.