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

A simple method for the determination of polyamines and their N-acetylated forms was developed using 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 calibration 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 concentration 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). Successful determination of the studied polyamines and their N-acetylated forms was performed on the saliva of 17 volunteers.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper
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