Abstract
Polyamines are ubiquitous active biogenic amines which contribute to basic cellular functions. Hence, their quantification in samples of diverse biological origins is essential for understanding how they function, especially in disease-relevant conditions. We present here a robust, high-throughput solid-phase extraction online coupled to a liquid chromatography-tandem mass spectrometry (SPE-LC/MS/MS) approach for the simultaneous quantification of eight polyamines in various biological samples. The polyamines include 1,3-diaminopropane, putrescine, cadaverin, N-acetyl-putrescine, spermidine, spermine, N(1)-acetyl-spermine, and l-ornithine. The novelty of the work is the use of two SPE columns online coupled to LC/MS/MS, which minimizes the sample pretreatment to a single derivatization step. The analysis is complete within 4min, making the method highly suitable for routine clinical analysis and high throughput screenings. The method was fully validated with serum samples. Dynamic ranges were 0.03 to 15μg/ml for ornithine and 1 to 500ng/ml for other polyamines, which cover physiological concentrations in serum samples. Lower limits of quantification (LLoQ) were found to be between 0.1 and 5ng/ml. As a proof of concept, we investigated gender differences in polyamine levels by analyzing the serum levels of 102 subjects.