Development and Validation of an LC Assay for Benzo[f]quinoline‐5,6‐dione and Identification of its Impurities by LC‐MS

Abstract

A new analytical method based on reversed‐phase high performanc liquid chromatography (HPLC) was developed and validated for the assay of benzo[f]quinoline‐5,6‐dione and the determination of its synthetic impurities by employing the method in liquid chromatography‐mass spectrometry (LC‐MS) with electrospray ionization (ESI) and photodiode array (PDA) UV detection. Separation was performed on a Waters Xterra C₁₈ (150 mm×4.6 mm, 5 µm) column. UV detection was performed at 262 nm. The results showed that benzo[f]quinoline‐5,6‐dione is eluted as a spectrally pure peak resolved from its impurities. 1H‐indeno[2,1‐b]pyridine‐2,9‐dion and benzo[f]quinoline are identified as the main impurities. The proposed method was extensively validated and satisfactory results were obtained in terms of linearity (r ² =0.9997) and precision (RSD<0.61%) in all cases. The limits of detection and quantification were 2 µg/mL and 20 µg/mL, with a 0.88% RSD, respectively.

Keywords:

• Benzo[f]quinoline‐5,6‐dione
• HPLC
• LC‐MS
• Method development
• Method validation
• Synthetic impurities

Acknowledgments

I would like to thank Jon Salmon and Phil Durban at Abbott Diabetes Care UK, Abbott Laboratories as well as Susan Tall at Abbott Diabetes Care Inc., California for reviewing this research for publication.