Analysis and Evaluation of Chiral Drugs in Biological Samples Using the Nexera UC-MS/MS System

Significance of the Topic

Chiral SFC-MS/MS offers rapid and high-resolution separation of enantiomeric drugs in biological matrices. This capability is crucial for pharmacokinetic studies, therapeutic drug monitoring, and quality control, where accurate quantification of individual enantiomers impacts drug efficacy and safety.

Study Objectives and Overview

This study aimed to develop and evaluate a supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS) method for enantiomeric analysis of omeprazole and rabeprazole in human plasma using the Nexera UC-MS/MS system. Column selection and mobile phase optimization were performed to achieve high selectivity and sensitivity for pharmacokinetic monitoring.

Methodology and Instrumentation

• Sample Preparation: Protein precipitation of plasma with acetonitrile and aqueous ammonia, followed by filtration and 3 μL injection.
• Chromatographic Conditions: CHIRALPAK IC-3 column (100×3.0 mm, 3 μm), supercritical CO2 with methanol modifier (A/B ratio 5/1 for omeprazole, 4/1 for rabeprazole), flow rate 3 mL/min, column temperature 40 °C, back pressure 10 MPa, BPR temperature 50 °C.
• Mass Spectrometry: LCMS-8050 triple quadrupole with ESI source in MRM mode. Transitions: m/z 346.1>198.1 for omeprazole, 359.9>150.1 for rabeprazole; make-up flow of methanol at 0.1 mL/min.

Main Results and Discussion

• Omeprazole: Enantiomers demonstrated baseline resolution. Calibration linearity (corrected by 1/[conc]2) yielded r2 values of 0.99996 (fast-eluting) and 0.99998 (slow-eluting). Repeatability at 2 μg/L showed RSD of 4.4% for both enantiomers; recovery rates at 10 μg/L were 101.1% and 100.5%.
• Rabeprazole: Similar chromatographic behavior allowed analysis with modified mobile phase composition and MRM settings. Linearity ranged r2 = 0.99996–0.99999 across 0.3–300 μg/L. Repeatability at 10 μg/L gave RSD of 1.8% and 2.4%; recoveries were 102.5% and 100.1%.

Benefits and Practical Applications

The developed SFC-MS/MS method delivers high-throughput analysis with excellent enantiomeric resolution, precision, and accuracy. It is directly applicable to pharmacokinetic profiling, bioequivalence studies, and routine QC of chiral drugs in clinical and pharmaceutical laboratories.

Future Trends and Potential Applications

• Expanding the methodology to a broader range of chiral pharmaceuticals.
• Integrating automated sample preparation and online coupling for increased throughput.
• Adoption of higher-pressure SFC and ultra-high performance columns to improve resolution and reduce analysis time.
• Applying data-driven optimization and AI algorithms for rapid method development.

Conclusion

The study confirms that the Nexera UC-MS/MS SFC-MS/MS platform, combined with the CHIRALPAK IC-3 column, is effective for accurate enantiomeric quantification of omeprazole and rabeprazole in plasma. The method exhibits robust linearity, repeatability, and recovery, establishing a reliable tool for chiral drug analysis in biomedical research and quality control.

Used Instrumentation

• Chromatograph: Nexera UC with SFC capabilities.
• Column: CHIRALPAK IC-3 (100 mm × 3.0 mm, 3 μm).
• Detector: Shimadzu LCMS-8050 triple quadrupole MS with ESI, MRM mode.

References

• Watabe Y., Hattori T., Iida T. Application News No. L517, Shimadzu Corporation, First Edition: Mar. 2017.
• Shimadzu Corporation. Application News No. L495.