Column selection approach for related substances determination of progesterone by high-performance liquid chromatography.

In this study, HPLC-MS/MS with triple-quadrupole tandem mass spectrometer was used to analyze the base-catalyzed thermal degradation derivative of progesterone in Chinese Pharmacopoeia. The full-scan spectrometry revealed that the degradation product was an isomer of progesterone, it was proposed as impurity M ((17α)-pregn-4-ene-3,20-dione) in European Pharmacopoeia, and both the derivative and progesterone were neutral molecules. Four kinds of chromatographic column characterization databases including PQRI database using Snyder/Dolan method, the USP chromatographic column database using the SRM 870 tests, the Tanaka/Euerby approach within the ACD program, and the Hoogmartens approach were compared. Combing the principles of column characterization databases with the system requirement of progesterone-related substances testing, a suitable PQRI-based method was finally set up for progesterone-related substances detecting. Our results indicated that PQRI system of Snyder/Dolan method was the most suitable system for related substances detection of progesterone.