Development of a method for the determination of polyamines including N-acetylated forms in human saliva via benzoylation and gas chromatography-mass spectrometry

Abstract

[EN]A simple method for the determination of polyamines and their N -acetylated forms was developed us- ing benzoyl chloride as derivatization reagent, and 1,6-diaminohexane as internal standard, followed by liquid-liquid extraction with ethyl acetate. The organic extract was injected in a gas chromatograph using a programmed temperature vaporizer and the determination and quantification was performed with a quadrupole mass spectrometer. There was no matrix effect with the proposed method, so internal cali- bration was used to quantify the corresponding derivatives. Good linear responses were obtained in the range from the limits of detection to 500 μg L −1 (50 μg L −1 for spermidine), with correlation coefficients varying from 0.9591 to 0.9968. The limits of quantification (S/ N = 10) ranged 1.0 –8.3 μg L −1 . Recoveries were found between 82 –117%, showing the good accuracy of the proposed method. Intra- and inter-day precision assays, expressed as relative standard deviation (RSD) were evaluated at two different concen- tration levels (low and high), showing values in the range of 2.4 –6.1% and 5.2 –9.0% for repeatability and reproducibility, respectively (6.9 –9.7% and 14.1 –14.6% for spermidine). Successfuldetermination of the studied polyamines and their N -acetylated forms was performed on the saliva of 17 volunteers.