title: Analysis of free, mono- and diacetylated polyamines from human urine by LC-MS/MS.
abstract: Polyamines are promising biochemical markers of cancer and many other pathophysiological conditions, and thus their concentrations in biological fluids are a matter of interest. However, since the concentrations of these compounds are low, their quantitation is typically based on methods requiring laborious sample preparation. Here we developed and validated an LC-MS/MS method to analyze simultaneously free (DAP, PUT, CAD, SPD, SPM) monoacetylated (AcPUT, AcCAD, N(1)AcSPD, N(8)AcSPD, N(1)AcSPM) and diacetylated (DiAcPUT, DiAcCAD, DiAcSPD, DiAcSPM) polyamines from human urine without the need for derivatization. Deuterium labeled polyamines were the internal standards for each analyte. Diluted urine samples spiked with internal standards were filtered through a strong anion exchange resin prior to LC-MS/MS analysis. The chromatographic separation of 14 polyamines was achieved in 12min on C18 column with 0.1% HFBA (v/v) as the ion-pairing agent and a water-acetonitrile gradient. Ionization was performed with positive electrospray ionization (ESI) and detection was with a triple quadrupole mass spectrometer with selected reaction monitoring. Calibration curves ranged from up to 5 to 10,000nM. The accuracy and precision of the method were determined using urine based quality control samples, and matrix effects were examined by using standard addition methods. This novel method is suitable for elucidating differences in urinary polyamine excretion in cancer patients and healthy humans.
authors.affiliation: School of Pharmacy, Biocenter Kuopio, University of Eastern Finland, Kuopio, Finland. Electronic address: Merja.Hakkinen@uef.fi.
authors.forename: Merja R
authors.initials: MR
authors.lastname: Häkkinen
authors.forename: Antti
authors.initials: A
authors.lastname: Roine
authors.forename: Seppo
authors.initials: S
authors.lastname: Auriola
authors.forename: Antti
authors.initials: A
authors.lastname: Tuokko
authors.forename: Erik
authors.initials: E
authors.lastname: Veskimäe
authors.forename: Tuomo A
authors.initials: TA
authors.lastname: Keinänen
authors.forename: Terho
authors.initials: T
authors.lastname: Lehtimäki
authors.forename: Niku
authors.initials: N
authors.lastname: Oksala
authors.forename: Jouko
authors.initials: J
authors.lastname: Vepsäläinen
mesh_terms: D000107:Acetylation; D000328:Adult; D000368:Aged; D002138:Calibration; D016022:Case-Control Studies; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011073:Polyamines / Q000652:urine*; D011786:Quality Control; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D011073:Polyamines
keywords: AcCAD; AcPUT; CAD; Cancer diagnostic markers; DAP; DiAcCAD; DiAcPUT; DiAcSPD; DiAcSPM; Liquid chromatography–tandem mass spectrometry; N(1)-(3-aminopropyl)butane-1,4-diamine trihydrochloride; N(1)AcSPD; N(1)AcSPM; N(8)AcSPD; N,N′-((butane-1,4-diylbis(azanediyl))bis(propane-3,1-diyl))diacetamide dihydrochloride; N,N′-(butane-1,4-diyl)diacetamide; N,N′-(pentane-1,5-diyl)diacetamide; N,N′-bis-(3-aminopropyl)butane-1,4-diamine tetrahydrochloride; N-(3-((4-((3-aminopropyl)amino)butyl)amino)propyl)acetamide trihydrochloride; N-(3-((4-acetamidobutyl)amino)propyl)acetamide hydrochloride; N-(3-((4-aminobutyl)amino)propyl)acetamide dihydrochloride; N-(4-((3-aminopropyl)amino)butyl)acetamide dihydrochloride; N-(4-aminobutyl)acetamide hydrochloride; N-(5-aminopentyl)acetamide hydrochloride; N-acetylated; PUT; Polyamines; Prostate cancer; SPD; SPM; Urine; butane-1,4-diamine dihydrochloride; pentane-1,5-diamine dihydrochloride; propane-1,3-diamine dihydrochloride