Detection of controlled substances in blood samples using the VeriSpray ion source with TSQ Altis MS for clinical research and forensic toxicology

Importance of the Topic

Rapid and reliable detection of controlled substances in blood is critical for clinical research and forensic toxicology. The rising abuse of opiates, stimulants, and designer drugs demands high-throughput, sensitive analytical methods to inform patient care, legal investigations, and public health strategies.

Study Objectives and Overview

This work aimed to develop a robust, sensitive, and reproducible PaperSpray-mass spectrometry workflow for detecting multiple illicit drugs in human blood. The focus was on creating a single quantitative assay capable of simultaneous measurement of six target analytes: cocaine, diazepam, fentanyl, hydrocodone, methamphetamine, and zolpidem.

• Establish calibration in dried blood spots with minimal sample preparation
• Optimize SRM transitions on a triple quadrupole mass spectrometer
• Evaluate method sensitivity, linearity, and throughput

Methodology and Instrumentation

Samples were prepared by spiking pooled human blood with isotopically labeled standards and depositing 6 µL spots onto VeriSpray sampling plates. Spots dried at room temperature or 40 °C before automated extraction and ionization. PaperSpray solvents of acetonitrile/acetone/water with 0.01% acetic acid were applied in controlled steps. The TSQ Altis triple quadrupole mass spectrometer operated in selected reaction monitoring (SRM) mode with time-dependent spray voltage, 0.7 Da resolution for Q1 and Q3, and a 0.8 s cycle time.

Instrumentation

• Thermo Scientific VeriSpray PaperSpray ion source with sampling plates (24 tips)
• Thermo Scientific TSQ Altis triple quadrupole mass spectrometer
• Thermo Scientific TraceFinder software version 4.1

Results and Discussion

Calibration curves for all analytes exhibited excellent linearity (R² > 0.98). Limits of detection ranged from 3 ng/mL for fentanyl to 68 ng/mL for methamphetamine, all within clinically relevant concentrations except for buprenorphine at 13 ng/mL. The automated workflow achieved a total analysis time of approximately two minutes per sample, combining extraction and detection without manual intervention.

Benefits and Practical Applications

• Fast sample turnaround (~2 min per dried blood spot)
• No extensive sample cleanup or chromatographic separation required
• Simultaneous quantitation of six drugs of abuse in one assay
• Reduced per-sample cost and increased laboratory throughput

Future Trends and Potential Applications

Advancements may include integration with high-resolution mass spectrometry for broader screening, expansion to additional biomarkers, and coupling with automated data analysis tools. The simplified workflow is poised for adoption in point-of-care testing, roadside screening, and large-scale epidemiological studies.

Conclusion

The VeriSpray PaperSpray-TSQ Altis method provides a rapid, sensitive, and reproducible platform for multi-analyte detection of controlled substances in blood. Its minimal sample preparation and high throughput address key needs in clinical research and forensic toxicology, supporting timely decision-making and evidence generation.

References

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