Rapid and Simultaneous Quantification of Polyamines in Human Plasma by LC–MS/MS After Pre-column Derivatization with N-(9-Fluorenylmethoxycarbonyloxy)succinimide

Abstract

Polyamines play an important role in growth and proliferations of cells, synthesis of proteins and nucleic acids. Thus, a sensitive analytical method for quantitative analysis of each polyamine in biological samples is a prerequisite for the further investigation of the application of polyamines in vivo. A high-performance liquid chromatography-tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of 1,3-diaminopropane, putrescine, cadaverine, spermidine and spermine in human plasma. The analytes were first extracted from plasma samples by protein precipitation procedure, and then derivatized by N-(9-fluorenylmethoxycarbonyloxy)succinimide with 1,6-diaminohexane as internal standard. The derivatives were separated on a Capcell core ADME reverse-phase column (2.7 μm, 2.1 mm i.d. ×50 mm) using a gradient program. Employing multiple-reaction monitoring, lower limit of quantification for polyamines ranged from 0.2 to 20 ng mL⁻¹, except spermidine and spermine from 2 to 200 ng mL⁻¹. The intra- and inter-day RSD for all polyamines were 0.9–13.9 % and 4.3–19.9 %, respectively. The proposed method was simple, effective and sensitive, and seems to be useful for the assessment of the polyamine status of cancer patients.