9-Fluoreneacetyl-tagged, solid-phase reagent for derivatization in direct plasma injection

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Accurate quantification of amphetamine in biological matrices is crucial for clinical and forensic toxicology.
• Existing methods for amphetamine analysis often involve complex sample preparation steps.

Purpose of the Study:

• To develop a simplified, sensitive, and direct injection method for amphetamine analysis in plasma.
• To introduce a novel resin-based solid-phase derivatization reagent for on-line pre-column derivatization.

Main Methods:

• A resin-based solid-phase derivatization reagent with a 9-fluoreneacetyl tag was synthesized and characterized.
• On-line, pre-column derivatization was performed by direct injection of diluted plasma into a sodium dodecyl sulfate-containing mobile phase.
• Amphetamine was trapped, derivatized at elevated temperature, separated by reversed-phase high-performance liquid chromatography, and detected by fluorescence.

Main Results:

• The developed method demonstrated good reproducibility and sensitivity for amphetamine analysis.
• The solid-phase reagent effectively facilitated on-line derivatization, simplifying the analytical procedure.
• Successful evaluation through single blind spiking analysis confirmed the method's reliability.

Conclusions:

• The combination of the solid-phase reagent and surfactant-containing mobile phase offers a sensitive and straightforward approach for direct injection analysis of biological fluids.
• This method significantly reduces sample preparation time and complexity for amphetamine determination.