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Analysis of underivatized polyamines by reversed phase liquid chromatography with electrospray tandem mass spectrometry.

Journal of pharmaceutical and biomedical analysis (2007-10-20)
Merja R Häkkinen, Tuomo A Keinänen, Jouko Vepsäläinen, Alex R Khomutov, Leena Alhonen, Juhani Jänne, Seppo Auriola
ABSTRACT

A reversed phase liquid chromatography-electrospray ionization-tandem mass spectrometric method (RP-LC-ESI-MS/MS) was developed to separate and detect polyamines with minimal sample pre-treatment and without any derivatization. Prior to MS/MS analysis, a complete chromatographic separation of polyamines was achieved by a linear gradient elution using heptafluorobutyric acid as a volatile ion-pair modifier, and signal suppression was prevented by post-column addition of 75% propionic acid in isopropanol to the column flow. The developed method was successfully applied to the identification of metabolites formed from N(1),N(12)-diethylspermine in the reaction catalyzed by recombinant human polyamine oxidase and to the detection of eight different polyamines in a standard mixture.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Silver heptafluorobutyrate, 97%
Sigma-Aldrich
Heptafluorobutyric acid, 98%
Supelco
Heptafluorobutyric acid, suitable for ion chromatography, ≥99.5% (GC)
Sigma-Aldrich
Heptafluorobutyric acid, ≥99.0% (GC)
Supelco
Heptafluorobutyric acid solution, 0.5 M in H2O, LiChropur, suitable for ion pair chromatography