AGILENT BOND ELUT CERTIFY AND CERTIFY II METHOD MANUAL

Importance of the topic

Forensic toxicology laboratories face the critical task of identifying and quantifying a wide variety of drugs of abuse and pharmaceuticals in complex biological fluids. Effective and reproducible sample preparation is essential to achieve accurate trace‐level detection, maintain sample integrity, and support regulatory and legal requirements.

Objectives and overview

This methods manual provides detailed workflows for Agilent Bond Elut Certify (strong cation‐exchange and C8) and Certify II (strong anion‐exchange and C8) mixed‐mode SPE products. It covers general sorbent chemistry, method optimization, solvent and buffer recipes, step‐by‐step extraction protocols for major drug classes (basic, acidic, neutral), and guidelines for method adaptation across sample types (urine, blood, plasma, meconium).

Methods and instrumentation

Protocols rely on three retention mechanisms—hydrophobic, polar, and ion‐exchange—matched for clean, reproducible recoveries. Key steps include cartridge conditioning with methanol and buffer, controlled sample loading, sequential washes to remove matrix interferences, drying steps to optimize solvent contact, and selective elution with alkaline or acidic organic solvents. Derivatization options (BSTFA, MSTFA, PFPA, etc.) prepare extracts for GC/MS, and alternative GC/FID or LC/MS workflows are described. Sample preparation hardware includes Vac Elut vacuum manifolds, 96‐well plate SPE systems, and various adapters for automation.

Used instrumentation

• Gas chromatographs with split/splitless inlets
• Mass spectrometers (GC/MS, GC/MSD, triple quadrupole LC/MS/MS)
• Liquid chromatographs (HPLC, UHPLC) with UV and MS detectors
• Mixed‐mode SPE cartridges and 96‐well plate formats
• Vacuum manifolds and adapters (Vac Elut, Vac Elut SPS 24, VersaPlate)
• Automated SPE platforms (Gilson ASPEC)
• pH meters and electrodes for buffer preparation

Main results and discussion

Validated methods demonstrate high recoveries, low matrix effects, and robust cleanup for a wide spectrum of drug analytes—amphetamines, opiates, benzodiazepines, stimulants, barbiturates, cannabinoids, and designer drugs—in multiple matrices. Critical control points include pH adjustment after hydrolysis, wash solvent selection for polar and nonpolar interferences, and cartridge drying times to maximize elution efficiency.

Benefits and practical applications

• Universal mixed‐mode sorbents simplify inventory and method development across drug classes
• Scalable formats (cartridge, 96‐well, VersaPlate) enable manual or automated high throughput
• Optimized for GC, GC/MS, HPLC, and LC/MS platforms without extensive revalidation
• Flexible sample volumes (0.5–5 mL) support trace‐level sensitivity for clinical, forensic, workplace, and research settings

Future trends and possibilities

Advances in ultra‐high‐pressure LC and fast GC, combined with more sensitive MS detectors, will drive lower detection limits and faster analyses. Emerging SPE sorbent chemistries and miniaturized formats will further streamline workflows. Integration with automated liquid handlers and 96‐well platforms will expand sample throughput and support multiplexed panels for new psychoactive substances and metabolic biomarkers.

Conclusion

Agilent Bond Elut Certify and Certify II SPE solutions deliver versatile, reliable, and reproducible sample preparation for forensic toxicology and related fields. This comprehensive manual equips laboratories with validated protocols, detailed solvent and buffer recipes, instrumentation guidelines, and optimization strategies to confidently detect and quantify drugs of abuse and pharmaceuticals in complex matrices.

References

• Chen X-H. Mixed-Mode Solid-Phase Extraction for the Screening of Drugs in Systematic Toxicological Analysis. University of Groningen; 1993.
• Hawks RL, Chiang CN, eds. Urine Testing for Drugs of Abuse. NIDA Research Monograph 73; 1986.
• McBay AJ. Analytical Aspects of Drug Testing. Wiley; 1989.
• Pierce Chemical Co. Catalog and Handbook; 1994.
• Sadee W, Beelen GCM. Drug Level Monitoring. Wiley; 1980.
• Simpson N, Van Horne KC. Sorbent Extraction Technology. Varian; 1993.
• UN International Drug Control Program. Recommended Methods for the Detection and Assay of Heroin and Cannabinoids; 1993.
• UN International Drug Control Program. Recommended Methods for the Detection and Assay of Cocaine, Amphetamine, Methamphetamine; 1993.