Quantitative Determination of Drugs Using Supercritical Fluid Chromatography with Triple Quadrupole Mass Spectrometry

Significance of the Topic

Supercritical fluid chromatography coupled with triple quadrupole mass spectrometry offers rapid separation of multiple drug classes in forensic toxicology while maintaining high sensitivity and precision. This approach addresses the need for efficient screening of amphetamines benzodiazepines morphine analogues and cannabinoids in disciplines such as doping control postmortem analysis and drug screening.

Objectives and Study Overview

This application note aimed to evaluate an analytical system comprising the 1260 Infinity Analytical SFC and 6460 triple quadrupole MS for quantitative analysis of 25 forensic relevant drugs. The study focused on method development calibration performance sensitivity and a demonstration using amphetamines in a spiked urine sample.

Methodology and Instrumentation

The chromatographic separation was achieved on a C8 stationary phase with a gradient of methanol containing formic acid and ammonium formate at a flow rate of 2 mL per minute and back pressure of 200 bar. Detection was performed in positive ion mode using dynamic multiple reaction monitoring. Key instrument components included the 1260 Infinity SFC control module isocratic pump binary pump degasser autosampler thermostatted column compartment high pressure diode array detector make up flow delivery and the 6460 triple quadrupole LC MS instrument equipped with a Jet Stream source. Standards were prepared from a forensic toxicology test mixture diluted to generate calibration points from 0.2 to 100 nanograms per milliliter. A urine sample was spiked at 100 nanograms per milliliter then diluted filtered and injected directly.

Main Results and Discussion

The optimized SFC method resolved all 25 analytes within five minutes with retention time relative standard deviations below 0.3 and peak area RSDs below 4. Calibration curves exhibited linearity coefficients above 0.994 across the range down to individual LOQs below 0.1 nanograms per milliliter and LODs below 0.03 nanograms per milliliter. Precision of concentration determinations was better than 3.5 and accuracy ranged from 95 to 110. In the spiked urine test the amphetamine class showed retention RSDs below 0.4 concentration RSDs below 3 and accuracy between 82 and 98. Comparative analysis of quantifier qualifier ion ratios confirmed the specificity of the method.

Benefits and Practical Applications

Rapid analysis time high sensitivity and robust quantification make this method ideal for high throughput forensic toxicology laboratories. The simplicity of a single injection covering multiple drug classes improves workflow efficiency in clinical toxicology doping control and law enforcement testing.

Future Trends and Potential Uses

Advancements in SFC stationary phases and expanded dynamic monitoring libraries will further extend the range of detectable substances and increase throughput. Integration with automated sample preparation and data processing software will enhance routine screening capabilities and support emerging applications in clinical diagnostics environmental analysis and metabolomics.

Conclusion

The combined use of supercritical fluid chromatography and triple quadrupole mass spectrometry delivers a powerful platform for rapid multiplexed drug quantification. This configuration meets the stringent requirements of forensic toxicology offering excellent sensitivity linearity precision and speed.

Reference

Stone PJW An Application Kit for Screening of Forensic and Toxicological Analytes using LC QQQ MS MS with Dynamic MRM Database Agilent Technologies application note publication 5990 4254EN 2016