NEAFS: Fast quantitative Forensic Analysis of THC and its Metabolites in Biological Samples using Captiva EMR- Lipid and LC/MSMS

Importance of the Topic

Accurate and rapid quantification of THC and its metabolites in biological matrices is critical for forensic investigations, workplace testing, and clinical research. Endogenous interferences such as lipids and phospholipids can compromise sensitivity, chromatographic performance, and instrument longevity. Streamlined sample preparation that efficiently removes these matrix components improves analytical robustness and throughput, meeting the growing demand for reliable toxicology data.

Objectives and Study Overview

This study evaluates a novel sample cleanup strategy—Captiva EMR-Lipid—for forensic quantification of THC, 11‐hydroxy-THC, and 11-nor-9-carboxy-THC in human plasma and whole blood using LC-MS/MS. Key goals include minimizing matrix effects, maximizing analyte recovery, and streamlining workflows relative to traditional protein precipitation and liquid-liquid extraction methods.

Methodology and Instrumentation

The workflow employs a single-step lipid removal cartridge (Captiva EMR-Lipid) in a pass-through format. Samples (100 µL plasma or whole blood) are mixed with 500 µL acidified acetonitrile, briefly vortexed or allowed to mix in situ, and drawn through the cartridge under low vacuum. A water/acetonitrile wash follows, then eluates are dried and reconstituted in methanol with formic acid. Quantitation is performed on an LC-MS/MS system monitoring MRM transitions for each analyte. Key instrumentation:

• Captiva EMR-Lipid cartridges
• High-performance LC coupled to triple quadrupole MS
• Heidolph Multi Reax® vortex mixer

Key Results and Discussion

Matrix removal performance was assessed by tracking 11 phospholipid transitions. Captiva EMR-Lipid eliminated over 99% of monitored phospholipids compared to protein precipitation, resulting in significantly reduced ion suppression. In plasma, recoveries for THC, THC-OH, and THC-COOH ranged from 90% to 116% across 0.5–100 ng/mL, with intra- and inter-day RSDs below 10%. Whole blood recoveries were similarly robust (92–113%), demonstrating the method’s consistency. The simplified protocol yielded cleaner chromatograms and extended column lifetimes by preventing lipid build-up.

Benefits and Practical Applications

Key advantages of the Captiva EMR-Lipid approach include:

• Rapid, one-step cleanup compatible with high-throughput workflows
• Consistently high analyte recoveries and low variability
• Substantial reduction of lipid-related ion suppression
• Broader applicability to diverse drug panels and lipophilic compounds

These benefits support forensic, clinical, pharmaceutical, and veterinary laboratories requiring reliable quantitation of cannabinoids and other mid- to late-eluting analytes.

Future Trends and Opportunities

Emerging forensic and clinical applications demand even greater sensitivity and multiplexing capacity. Integrating enhanced matrix removal with automated sample handlers and miniaturized LC-MS platforms could further accelerate turnaround and reduce solvent consumption. Expanding sorbent chemistries to target additional endogenous interferences—such as metabolic conjugates or polar lipid classes—will broaden applicability. Advances in data analytics and machine learning promise to optimize method parameters in real time, enhancing throughput and robustness.

Conclusion

The Captiva EMR-Lipid cartridge delivers a streamlined, one-step cleanup that effectively removes lipids, preserves target analytes, and simplifies sample preparation for forensic quantitation of THC and its metabolites. This approach outperforms traditional cleanup methods by improving chromatographic clarity, reducing ion suppression, and maintaining high recovery and precision. It offers a robust solution for laboratories seeking to enhance throughput and data quality.

Reference

Deckers C. Fast quantitative forensic analysis of THC and its metabolites in biological samples using Captiva EMR-Lipid and LC/MSMS. Agilent Technologies Application Note.