Usability Study of a New HPLC, a New Tandem MS and a New Data Processing Software

Significance of the Topic

High-throughput clinical and industrial laboratories demand analytical platforms that deliver consistent performance, minimal downtime, and streamlined data management. Innovations in chromatography, mass spectrometry, and software integration can greatly enhance productivity, reduce manual intervention, and ensure reliable results over extended sample runs.

Objectives and Study Overview

This usability study aimed to evaluate the combined performance of a new two-channel HPLC with online sample cleanup, a tandem mass spectrometer capable of rapid polarity switching, and dedicated data processing software under routine clinical-style workloads. Key objectives included:

• Assessing long-term retention time and concentration precision over 2 000 injections.
• Evaluating inter-instrument reproducibility across three identical units.
• Measuring improvements in sample throughput and MS utilization.
• Demonstrating software-enabled workflow flexibility and audit-ready reporting.

Methodology and Instrumentation

The experimental workflow combined:

• HPLC System: A dual-channel configuration featuring TurboFlow™ columns for automated online cleanup and analytical columns (Cyclone-P™ or Accucore™ aQ; 50×2.1 mm, 2.6 µm) operated at 25–30 °C.
• Chromatographic Conditions: Mobile phases of 10 mM ammonium formate with 0.05% formic acid in water and methanol, plus an organic wash (isopropanol:acetonitrile:acetone 45:45:10) for channel equilibration.
• Mass Spectrometer: Tandem MS with heated electrospray ionization (HESI) probe and atmospheric pressure chemical ionization (APCI) probe in polarity switching mode. Selected reaction monitoring transitions were optimized for four test compounds (reserpine, chloramphenicol, testosterone, estradiol) and their deuterated internal standards.
• Data Processing: ClinQuan MD™ software provided role-based access (technician, supervisor, director), real-time quantitation, ion ratio monitoring, and audit-trail record keeping.

Main Results and Discussion

Continuous operation over 100 hours (2 000 injections) yielded:

• Retention Time Precision: 0.85% RSD (Alprazolam across two channels, n=2 000).
• Concentration Precision: 1.49% RSD for Alprazolam; inter-instrument concentration RSDs of 3.87% (reserpine), 6.61% (chloramphenicol), 3.48% (testosterone), and 8.08% (estradiol), based on n=120 replicates across three units.
• Ion Ratio Stability: Confirming-to-quantifying ion ratios maintained <3% RSD over the full injection set.
• Throughput Gains: Dual-channel operation eliminated mass spectrometer idle time during chromatographic steps, effectively doubling sample throughput compared to a single-channel system.
• Software Performance: ClinQuan MD streamlined data review, enabled rapid reprocessing, and secured audit-ready documentation without manual reporting steps.

Benefits and Practical Applications

The integrated platform delivers:

• High precision and reproducibility for routine clinical assays.
• Extended continuous run capability, reducing downtime for column cleaning or manual intervention.
• Enhanced system utilization through multichannel chromatographic sequencing.
• Automated data processing and compliance features to support regulatory requirements in QA/QC and clinical diagnostics.

Future Trends and Potential Applications

Advancements building on this work may include:

• Further miniaturization and multiplexing of chromatographic channels to boost throughput in large-scale screening.
• Integration of machine-learning algorithms for adaptive method optimization and predictive maintenance.
• Cloud-based data management for centralized monitoring across multiple laboratory sites.
• Expansion to biomarker discovery, therapeutic drug monitoring, and environmental testing workflows.

Conclusion

This usability study demonstrates that a dual-channel HPLC with online cleanup, a polarity-switching tandem MS, and dedicated processing software can achieve stable, precise, and highly efficient performance over thousands of injections. The combined system offers significant advantages for high-volume clinical and industrial laboratories, supporting robust method execution, regulatory compliance, and maximized instrument utilization.