Quantification of residual organic bases in an active pharmaceutical ingredient using mixed-mode chromatography and UV detection

Abstract

The use of organic bases is ubiquitous in chemical synthesis, yet quantifying these compounds with traditional HPLC methodologies is often hampered by poor peak shape, low retention, and limited UV absorption. When employed in the manufacture of an active pharmaceutical ingredient (API), these compounds must be controlled to levels that are safe for human consumption, requiring robust analytical methods with sufficiently low quantification limits. This work describes the development of an HPLC method for the quantification of imidazole and 1,8-Diazabicyclo[5.4.0]undec‑7-ene (DBU) in an API using mixed-mode chromatography. Through control of the pH and organic modifier gradients, the retention of the basic analytes and API can be tuned independently to achieve desirable retention and sensitivity for each compound. The resulting HPLC method exhibits good performance in linearity, accuracy, sensitivity, specificity, and solution stability. Notably, these conditions avoid more complex detection modalities, such as mass spectrometry, while maintaining a system pressure below 400 bar, making the method compatible with a broad range of instruments. This approach to mixed-mode chromatography method development could be extended to different organic bases in the presence of complex molecules to fit the needs of projects in an academic or industrial environment.

Keywords: Method Development; Mixed-mode chromatography; Organic base; Pharmaceuticals.