Fast and easy separation of 23 drugs of abuse using the Accucore Biphenyl Column

Importance of the Topic

Urgent need for reliable methods to quantify drugs of abuse in biological matrices supports forensic, clinical and research applications. Rapid and selective separation of multiple analytes enhances laboratory throughput and data confidence.

Study Objectives and Overview

This protocol demonstrates a fast UHPLC-MS/MS approach for separating 23 drugs of abuse, including challenging isobaric opioid pairs, from diluted human urine in under four minutes using a biphenyl stationary phase.

Methodology

• Sample Preparation: Dilution of 100 µL urine with 900 µL 0.1 % formic acid in water.
• Chromatography:
  • Column: 50 × 2.1 mm, 2.6 µm Accucore Biphenyl at 45 °C.
  • Mobile Phase A: 0.1 % formic acid in water; B: 0.1 % formic acid in methanol.
  • Gradient: 95 % A to 0 % A over 4.0 min, hold to 4.6 min, re-equilibrate to 95 % A by 5.5 min.
  • Flow Rate: 0.75 mL/min; Injection Volume: 2 µL.
• Detection: Heated electrospray ionization with multiple reaction monitoring.

Main Results and Discussion

All 23 analytes achieved baseline separation within four minutes, including isobaric opioids such as hydromorphone versus morphine and oxycodone versus naltrexone. Peak shapes were sharp and retention times consistent. Lower recovery for buprenorphine was attributed to vial adsorption and mitigated by using high-purity glass vials.

Benefits and Practical Applications

• High-throughput urine screening in forensic and clinical labs.
• Robust resolution of structurally similar opioid isobars.
• Minimal sample preparation reduces analysis time.
• Adaptable to complex matrices or lower detection thresholds with additional cleanup.

Future Trends and Opportunities

Future work may expand the analyte panel, integrate automated solid phase extraction, and enhance sensitivity for trace-level detection. The biphenyl phase could also find applications in environmental and food contaminant analysis.

Conclusion

The described UHPLC-MS/MS method using an Accucore Biphenyl column delivers rapid, efficient and reproducible separation of 23 drugs of abuse in urine. Its combination of speed, selectivity and robustness makes it ideal for routine analytical workflows.

Used Instrumentation

• Thermo Scientific Vanquish Horizon UHPLC System
• Thermo Scientific TSQ Quantiva Triple Quadrupole Mass Spectrometer
• Thermo Scientific Accucore Biphenyl Column, 50 × 2.1 mm, 2.6 µm
• Thermo Scientific Chromacol GOLD grade vials

References

1. CoE Application Scientists, Thermo Fisher Scientific, Drugs of abuse in human plasma using SOLAμ solid phase extraction, Application Brief 22007.
2. Kean Woodmansey et al., Fast and easy separation of 23 drugs of abuse including high stable resolution of isobaric opioids from human urine by UHPLC-MS/MS, Technical Note 21883.