Chiral purity assay for Flindokalner using tandem mass spectrometry: method development, validation, and benchmarking

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis
• Mass Spectrometry

Background:

• Chiral purity is critical for drug efficacy and safety, particularly for neuroprotective agents like Flindokalner.
• Existing chromatographic methods for chiral purity determination can be time-consuming.
• Development of rapid and accurate analytical techniques is essential for pharmaceutical development.

Purpose of the Study:

• To demonstrate the application and validation of a mass spectrometry (MS/MS) method for quantitative chiral purity determination.
• To analyze the chiral purity of Flindokalner, a drug candidate for post-stroke neuroprotection.
• To establish MS/MS as a viable alternative to chromatographic techniques for chiral analysis.

Main Methods:

• Quantitative chiral purity determination using tandem mass spectrometry (MS/MS) and the kinetic method.
• Validation of the MS/MS method.
• Benchmarking MS/MS against chiral high-performance liquid chromatography with ultra-violet detection (LC/UV) and achiral high-performance liquid chromatography with circular dichroism detection (LC/CD).

Main Results:

• The MS/MS method achieved rapid chiral quantification of Flindokalner with a 3-minute run time per sample.
• Accuracy and precision of the MS/MS method were comparable to established LC/UV and LC/CD techniques.
• The kinetic method in MS/MS proved effective for gas-phase chiral determinations.

Conclusions:

• Mass spectrometry, utilizing the kinetic method, provides a rapid and accurate alternative for quantitative chiral purity determination.
• This MS/MS method is particularly advantageous for high-throughput screening in pharmaceutical research.
• The validated method supports the development of Flindokalner and similar chiral drug candidates.