Agilent LC Triple Quadrupole Clinical Research Compendium

Importance of the topic

Liquid chromatography coupled to triple quadrupole mass spectrometry is a cornerstone technique in clinical research and routine analysis due to its unmatched selectivity, sensitivity, and throughput.

Objectives and Overview of the Compendium

This compendium collects application notes demonstrating Agilent Triple Quadrupole LC/MS systems in hormone analysis, therapeutic drug monitoring, vitamin D metabolism, proteomics, metabolomics, and productivity enhancement.

Instrumentation Used

• Agilent 1290 Infinity II and 1260 Infinity II Prime LC systems
• Agilent Ultivo, 6460, 6470, 6495 and 6495C triple quadrupole mass spectrometers with Jet Stream or ESI sources
• Columns: Poroshell C18/PFP, Eclipse Plus C18, ZORBAX RRHD C18
• Sample cleanup: Captiva EMR-Lipid, NDLipids, BondElut (PBA, Plexa), SimpliQ SPE, StreamSelect LC/MS multiplexing

Main Results and Discussion

• Hormone quantification: Progesterone metabolites, free testosterone, catecholamines, and 13 steroid hormones in serum achieved baseline separation, LOQs in the low ng/L to pg/mL range, and precision (RSD) <10%
• Therapeutic drug monitoring: Fast LC/MS/MS assays for barbiturates and THC-A in urine, melatonin in plasma, and immunosuppressants (cyclosporine A, everolimus, sirolimus, tacrolimus) in blood, with sub-minute run times and automated online cleanup
• Vitamin D: 25-OH D2 and D3 analysis in serum using Captiva EMR-Lipid to remove lipids and phospholipids, yielding recoveries >90%, accuracy 90–110%, and linearity (R2>0.99)
• Proteomics & Metabolomics: Standard-flow LC/TQ quantified plasma peptides with LLOQ down to amol/µL and excellent reproducibility; enantiomeric separation of D/L-2-hydroxyglutarate and α-ketoglutarate by DATAN derivatization and LC/MS/MS
• Productivity enhancement: StreamSelect LC/MS system multiplexed up to four parallel LC streams on a single mass spectrometer, increasing throughput by up to 4× without compromising sensitivity or reproducibility
• Cannabinoid profiling: Six cannabinoids in urine separated in under 80 seconds per stream, with 3.7× throughput gain using four-stream multiplexing, maintaining linearity (5–5000 ng/mL) and RSD <3%

Benefits and Practical Applications

These workflows deliver rapid turnaround, high selectivity for isobaric analytes, low LLOQs suitable for endogenous compounds, robust performance in complex matrices, and simplified operation for nonexpert users, supporting clinical research, QA/QC, and high-throughput screening.

Future Trends and Opportunities

• High-throughput multiplexed LC/MS for large clinical cohorts
• Adoption of standard-flow proteomics for routine biomarker measurement
• Advanced sample cleanup chemistries for extended robustness
• Automation and AI-driven data processing for real-time QC and method optimization

Conclusion

The Agilent Triple Quadrupole LC/MS application compendium highlights versatile, high-performance methods across key clinical research areas. Demonstrated workflows ensure sensitivity, precision, and significant productivity gains, paving the way for broader adoption in clinical and industrial laboratories.

Reference

• Peters KE, et al. Validation of a protein biomarker test for predicting renal decline in type 2 diabetes: the Fremantle Diabetes Study Phase II. J Diabetes Complications. 2019;107406.