Agilent Ultivo ESI for High-Throughput Detection of Drugs in Urine and Serum

Significance of the Topic

Forensic toxicology requires rapid and reliable detection of drugs in biofluids to support investigations and health monitoring. High-throughput methods with low detection limits are critical for laboratories handling large sample volumes.

Objectives and Study Overview

This application note details a seven-minute LC/MS/MS method for the simultaneous quantitation of 68 drugs of abuse in urine and serum using the Agilent Ultivo triple quadrupole system coupled with a 1290 Infinity II LC. The aim was to achieve limits of quantitation at or below 10 ng/mL with robust precision and linearity across a broad concentration range.

Methodology and Sample Preparation

The workflow employs dynamic multiple reaction monitoring (dMRM) for targeted detection. Serum samples undergo protein precipitation with cold acetonitrile, centrifugation, and dilution, while urine samples are simply diluted tenfold with water. Calibration curves spanning 0.1 to 1000 ng/mL were prepared in matrix. Data acquisition utilized dMRM windows tailored to each analyte’s retention time, and quantitation was performed with Agilent MassHunter Quantitative Analysis 10.1’s Quant-My-Way feature to expedite review.

Instrumentation

• Agilent 1290 Infinity II LC system: high-speed pump (G7120A), multisampler with cooler (G7167B), multicolumn thermostat (G7116B)
• Agilent Ultivo triple quadrupole LC/MS with electrospray ionization source (G1948B)

Main Results and Discussion

• All 68 analytes demonstrated limits of quantitation ≤10 ng/mL; numerous compounds reached 0.1 ng/mL
• Precision at the LOQ showed %RSD below 20% for ten replicate injections, with most values between 5% and 15%
• Calibration curves were linear (R2 > 0.98) across up to four orders of magnitude
• Chromatographic performance using a Poroshell 120 EC-C18 column at 55 °C and a seven-minute gradient yielded sharp, well-resolved peaks
• Compared to an earlier Agilent 6420 platform, Ultivo provided at least twofold sensitivity improvements for 97% of overlapping analytes, and tenfold or greater for 50%

Benefits and Practical Applications

The method offers exceptional throughput and minimal sample preparation, reducing turnaround time. The compact, stackable Ultivo design, along with early maintenance feedback and VacShield technology, enhances operational uptime. This approach is ideal for high-volume forensic and clinical toxicology laboratories requiring routine screening and quantitation of diverse drug classes.

Future Trends and Potential Applications

Future developments may encompass automated sample preparation, expansion to additional analytes and matrices (oral fluid, tissue), integration with high-resolution mass spectrometry, and advanced data analytics. These enhancements could further improve sensitivity, selectivity, and workflow efficiency for toxicology screening, compliance testing, and therapeutic drug monitoring.

Conclusion

The Agilent Ultivo LC/TQ method delivers a rapid, sensitive, and precise platform for multi-analyte drug detection in urine and serum with minimal sample preparation and high throughput. Its robust performance and efficient design meet the rigorous demands of modern forensic and clinical testing environments.

Reference

1. Sosienski, T.; Hitchcock, J. Robustness of the Agilent Ultivo Triple Quadrupole LC/MS with Standard ESI Ion Source for High-Throughput Testing of Drugs in Serum. Agilent Technologies Application Note 5994-0737EN, 2019.
2. Stone, P.; McCann, K. Comprehensive LC/MS Analysis of Illicit and Pain Management Drugs Including Their Metabolites in Urine. Agilent Technologies Application Note 5991-1667EN, 2013.