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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.