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Urine polyamines determination using dansyl chloride derivatization in solid-phase extraction cartridges and HPLC
C Molins-Legua1, P Campíns-Falcó, A Sevillano-Cabeza
1Departament de Química Analítica, Facultad de Química, Universitat de Valencia, Spain.
The Analyst
|December 22, 1999
Summary
Solid phase extraction using C18 cartridges offers an efficient method for derivatizing polyamines with dansyl chloride. This technique enhances analysis time and sensitivity for polyamine determination in biological samples.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Biogenic amines, including putrescine, cadaverine, spermidine, and spermine, are crucial biomarkers.
- Accurate quantification of polyamines is essential for clinical diagnostics.
Purpose of the Study:
- To develop and optimize a solid-phase extraction (SPE) method for dansyl chloride derivatization of polyamines.
- To apply the developed method for polyamine determination in human urine samples.
Main Methods:
- Testing various SPE cartridge materials, with C18 showing optimal analyte retention.
- Optimizing reaction conditions: pH 12 buffer, 2 mM dansyl chloride, room temperature, and 30 min reaction time.
- Utilizing High-Performance Liquid Chromatography (HPLC) with 1,7-diaminoheptane as an internal standard for quantification.
Main Results:
- C18 cartridges provided the highest retention for the tested polyamines.
- Optimal derivatization and extraction conditions were established.
- The method achieved detection limits of 10 ng mL⁻¹, improved to 2 ng mL⁻¹ after extract concentration.
- Accurate and precise quantification of polyamines in urine samples from healthy controls and cancer patients was demonstrated.
Conclusions:
- Solid-phase extraction offers advantages over solution-phase derivatization, including reduced analysis time and improved stability of dansyl derivatives.
- The developed SPE method is robust, sensitive, and suitable for automated polyamine analysis.
- This method provides a valuable tool for clinical research and diagnostics involving polyamine levels.