Clam- SPE on line- UHPLC-MS/MS : Fully automatic, simultaneous and quickly quantification of drugs of abuse in blood, plasma and urine

Significance of the Topic

The reliable and rapid quantification of drugs of abuse in blood, plasma and urine is essential for forensic, clinical and workplace testing. Traditional sample preparation methods are time-consuming, labor-intensive and prone to variability. Automating the process enhances throughput, reduces human error and conserves solvents, meeting increasing analytical demands.

Objectives and Overview

This study presents a fully automated on-line solid-phase extraction (SPE) coupled with ultra high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) for simultaneous detection and quantification of over 40 drugs of abuse and three major cannabinoids. The workflow aims to deliver a single-run, 10-minute analysis applicable to blood, plasma and urine with a unified calibration strategy.

Methodology and Instrumentation

The automated workflow employs:

• Shimadzu Clam-2000 module for protein precipitation, filtration and dilution directly from primary collection tubes.
• On-line SPE with mixed-mode exchange cartridges for sample cleanup.
• UHPLC Nexera XR system for chromatographic separation on a C18 column.
• Shimadzu LCMS-8060 triple quadrupole MS operating in MRM mode (two or three transitions per compound) for sensitive detection.
• Calibration performed in blood across 5–250 ng/mL for stimulants, opioids and metabolites, and 0.5–25 ng/mL for cannabinoids.

Main Results and Discussion

Linearity of calibration curves exceeded R² = 0.99. Limits of quantification were 10 ng/mL for most analytes and 1 ng/mL for cannabinoids (11-OH-THC, THC-COOH, THC). Calibration accuracy ranged from 85 % to 115 %, with ion-ratio stability within 30 %. Repeatability across blood, plasma and urine quality controls was below 15 %. The 10-minute run, including injector and SPE rinsing, achieved negligible carry-over and excellent robustness. A single blood-based calibration curve was successfully applied to plasma and urine matrices.

Benefits and Practical Applications

• High throughput: under 10 minutes per sample for >43 analytes in three biological fluids.
• Reduced manual handling: minimal intervention decreases contamination risk.
• Lower solvent consumption and cost savings through integrated sample cleanup.
• Consistent performance: unified calibration simplifies laboratory workflow.
• Applicability: suitable for forensic toxicology, emergency screening and routine monitoring.

Future Trends and Applications

Future developments may include adaptation of the Clam-2000/SPE on-line system to saliva collections (e.g., Floqswab® and Quantisal®), expansion of the target panel to emerging psychoactive substances, integration with laboratory information systems for end-to-end automation, and miniaturized or field-deployable variants to support point-of-care toxicology.

Conclusion

The described automated SPE-UHPLC-MS/MS method offers rapid, sensitive and simultaneous quantification of a broad range of drugs of abuse and cannabinoids in blood, plasma and urine. Its robust performance, minimal sample handling, single calibration approach and short analysis time make it a powerful tool for modern toxicological laboratories.

References

WO 2016/098169 A1 (Clam-2000 on-line sample preparation patent)