ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF OMADACYCLINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH MASS SPECTROMETRY

High-Performance Liquid Chromatography coupled with Mass Spectrometry (HPLC-MS) is a powerfulanalytical technique used to separate, identify, and quantify individual components within complexmixtures. The system integrates two distinct instruments: the HPLC, which performs the separation, andthe mass spectrometer, which provides precise identification and quantification based on mass-to-charge(m/z) ratios.In the HPLC component, a diluted sample—such as an extract from mussels or seaweed—isinjected into a chromatographic column packed with microscopic beads (typically 1.8–5 μm insize). These beads vary in composition depending on the separation requirements. As the samplepasses through the column, its individual components interact differently with the stationary phase(the beads) based on their polarity. A mobile phase—usually a mixture of solvents like water andacetonitrile—is pumped through the column in varying ratios. Compounds with similar polarity tothe column material are retained longer, while those with greater affinity to the solvent elute morequickly.