Simultaneous Quantitative Determination of Five Process Relevant Impurities in Menatetrenone

Menatetrenone, a form of vitamin K (MK-4) is used for various pharmaceutical and agrochemical applications. Monitoring and control of process impurities is crucial in the manufacturing of pharmaceutical-grade Menatetranone. In this article, we report a gas chromatography (GC)-based analytical method for the detection and quantification of trace levels of five process-related impurities in MK-4 originating from the starting materials and reagents. A GC-flame ionization detector (GC-FID)-based method is presented for simultaneous quantitative estimation of five structurally diverse compounds, some of which are conventionally analyzed by liquid chromatography. After a series of method development trials, an optimized method with a mid-polar column dura bond-624 (6% cyanopropyl/phenyl and 94% polydimethylsiloxane) with initial column oven temperature 150 degrees C, detector temperature 280 degrees C was finalized for the separation of these impurities. The developed GC-FID-based method was validated for specificity, the limit of detection (LOD), the limit of quantification (LOQ), linearity, range, precision, accuracy, and robustness as relevant to the International Council for Harmonization guidelines. The method was found to be highly sensitive for all the constituents with an LOD ranging from 0.05 to 0.43 pg and a limit of quantification of 0.17-1.41 pg, as estimated by the signal-to-noise ratios. This is the first report of an analytical method for the simultaneous quantitative determination of five process impurities of MK-4.