Drugs of Abuse in Urine by SAMHSA Protocol

Significance of the Topic

Confirmatory testing for drugs of abuse in urine is a cornerstone of workplace and clinical toxicology programs. Accurate identification and quantification of opioids and other controlled substances support legal defensibility, ensure regulatory compliance under SAMHSA guidelines, and protect public health.

Aims and Overview of the Study

This application note presents a validated gas chromatography–mass spectrometry (GC–MS) method for confirmatory analysis of codeine, morphine and the heroin-specific metabolite 6-acetylmorphine (6-AM) in human urine. The protocol meets SAMHSA requirements for sensitivity, selectivity and reproducibility at defined cutoff levels.

Methodology

The procedure comprises seven key steps:

• Addition of deuterated internal standards to 1–5 mL urine aliquots.
• pH adjustment and enzyme or acid hydrolysis to cleave glucuronide conjugates for opiates and cannabinoids.
• Solid phase extraction (SPE) using a polymeric C18 cartridge to isolate target analytes.
• Evaporation of eluates to dryness and derivatization with either pentafluoropropionic anhydride (PFPA) or BSTFA + 1% TMCS.
• Reconstitution in organic solvent and injection (1–3 µL) into a GC–MS system operating in selected ion monitoring (SIM) mode.
• Quantitation based on primary ion abundance and confirmation via two additional ions, with ion ratio acceptance ±20%.
• Data processing and reporting facilitated by TurboMass software, providing three-ion-ratio overlays for each analyte.

Instrumentation Used

• PerkinElmer Clarus 680 GC with pressure-pulsed splitless injection and Elite-5 capillary column (12 m × 0.20 mm, 0.33 µm).
• Helium carrier flow of 2 mL/min, oven program from 100 °C to 310 °C at 20 °C/min.
• PerkinElmer SQ8 MS in electron ionization (EI) mode using a 270 L/s turbomolecular pump.
• Data acquisition in SIM with 20–30 ms dwell time per ion.

Main Results and Discussion

The method achieved an LOQ of 20 ng/mL and an LOD below 2 ng/mL for each analyte from 1 mL urine. Linearity was demonstrated from 20 to 20 000 ng/mL with R² > 0.999. Representative chromatograms and ion ratio overlays confirm peak identity and compliance with SAMHSA cutoff criteria (2000 ng/mL for codeine and morphine; 10 ng/mL for 6-AM).

Benefits and Practical Applications

• High sensitivity and specificity suitable for forensic, clinical and research laboratories.
• Fast throughput enabled by a short GC column, rapid oven cycling and autosampler rinsing.
• Robustness ensured by deuterated internal standards and strict ion ratio monitoring.
• Legally defensible reporting through automated three-ion-ratio reports.

Future Trends and Opportunities

Advances in micro-extraction techniques and ultrafast GC–MS instrumentation may further reduce sample preparation time and increase throughput. Integration with high-resolution mass spectrometry could expand multiplexed assays for emerging novel psychoactive substances. Automation of hydrolysis and SPE steps promises greater reproducibility and reduced labor.

Conclusion

The described GC–MS method provides a reliable confirmatory assay for key opioids in urine, meeting SAMHSA regulatory requirements and delivering rapid, sensitive results. Its adaptability and robust performance make it an excellent choice for a wide range of toxicology applications.

References

• Baselt RC. Disposition of Toxic Drugs and Chemicals in Man, 8th Ed. Biomedical Publications; 2008.
• Mandatory Guidelines for Federal Workplace Drug Testing Programs. Fed Reg. 73:71857 (Nov 25, 2008).
• Mandatory Guidelines for Federal Workplace Drug Testing Programs. Fed Reg. 75:22809 (Apr 30, 2010).
• Pierce Catalog. Rockford, IL.