Analysis of underivatized polyamines by reversed phase liquid chromatography with electrospray tandem mass spectrometry.
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.
Häkkinen
Merja R
MR
Department of Biosciences, Laboratory of Chemistry, University of Kuopio, PO Box 1627, FI-70211 Kuopio, Finland. merja.hakkinen@uku.fi
Keinänen
Tuomo A
TA
Vepsäläinen
Jouko
J
Khomutov
Alex R
AR
Alhonen
Leena
L
Jänne
Juhani
J
Auriola
Seppo
S
D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D015394:Molecular Structure; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry; D011073:Polyamines / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014835:Volatilization; D000097667:Polyamine Oxidase
D005466:Fluorocarbons; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011073:Polyamines; D011994:Recombinant Proteins; D000097667:Polyamine Oxidase; C033094:perfluorobutyric acid