Rapid Screening Method for Statins Using an Advanced UHPLC Column and System

Significance of the Topic

The rapid and reliable analysis of statin drugs is critical in pharmaceutical quality control, clinical laboratories, and drug screening applications. High-throughput methods enable better process efficiency, cost reduction, and faster decision making in routine assays involving multiple analytes.

Objectives and Study Overview

This work demonstrates a fast screening method for nine statins using the Thermo Scientific Hypersil GOLD VANQUISH 1.9 µm UHPLC column coupled with the Vanquish UHPLC system. The study explores non-linear gradient capabilities and evaluates the impact of increased flow rates and backpressures on separation performance and assay throughput.

Methodology and Instrumentation

Sample Preparation:

• Primary solutions of nine statins at 1 mg/mL in methanol
• Working solutions at 100 µg/mL by dilution in water
• Vials labeled via Virtuoso Vial Identification System

UHPLC Conditions:

• Column: Hypersil GOLD VANQUISH, 1.9 µm, 100 × 2.1 mm
• Mobile Phase A: 0.1% formic acid in water
• Mobile Phase B: 0.1% formic acid in acetonitrile
• Column temperature: 40 °C with eluent pre-heating
• Injection volume: 1 µL; Detection wavelength: 240 nm
• Flow rates evaluated: 0.5, 1.0, 1.2, and 1.4 mL/min
• Gradient profiles: linear versus non-linear curves (curve value = 8)

Instrumentation:

• Vanquish UHPLC System: System Base, Binary Pump H, Split Sampler HT, Column Compartment H, Active Pre-heater, Diode Array Detector HL, LightPipe flow cell
• Chromatography Data System: Thermo Scientific Dionex Chromeleon 7.2 SR2

Main Results and Discussion

Non-linear gradient at 0.5 mL/min improved resolution of critical fluvastatin and atorvastatin peaks compared to a linear gradient. Increasing flow rates up to 1.4 mL/min while maintaining gradient slope kept peak resolution within acceptable USP criteria, despite backpressures rising to 1,334 bar.

Resolution Performance (1.0–1.4 mL/min):

• Amlodipine: 2.28–2.11
• Ezetimibe: 24.31–20.98
• Fluvastatin: 10.87–9.87
• Atorvastatin: 1.69–1.41
• Lovastatin: 7.34–5.32
• Simvastatin: 6.77–4.68

Precision: Twelve replicate injections at 1.2 mL/min non-linear method yielded retention time RSD ≤ 0.20% for all analytes, underscoring system stability and reproducibility.

Benefits and Practical Applications

• Method time under three minutes at maximum flow rates
• Baseline resolution of critical statin pairs
• High retention time precision (RSD ≤ 0.2%)
• Increased sample throughput through non-linear gradient optimization

Future Trends and Potential Applications

Advances in UHPLC columns and high-pressure systems will further accelerate multi-compound screening. Emerging areas include:

• Automation and AI-driven method development for gradient optimization
• Ultra-high pressure (> 1500 bar) workflows for faster analyses
• Integration with mass spectrometry for enhanced sensitivity and selectivity
• Miniaturization and multiplexing for clinical and bioanalytical platforms

Conclusion

The combination of the Hypersil GOLD VANQUISH UHPLC column with the Vanquish UHPLC system and non-linear gradient capabilities enables rapid, high-resolution screening of statins. This approach supports sub-three-minute assays with excellent reproducibility and significant throughput gains.

References

1. Hillbeck D. Application Note AN21496. Thermo Fisher Scientific; 2016.