Polyamines in biological samples: rapid and robust quantification by solid-phase extraction online-coupled to liquid chromatography-tandem mass spectrometry.
Polyamines are ubiquitous active biogenic amines which contribute to basic cellular functions. Hence, their quantification in samples of diverse biological origins is essential for understanding how they function, especially in disease-relevant conditions. We present here a robust, high-throughput solid-phase extraction online coupled to a liquid chromatography-tandem mass spectrometry (SPE-LC/MS/MS) approach for the simultaneous quantification of eight polyamines in various biological samples. The polyamines include 1,3-diaminopropane, putrescine, cadaverin, N-acetyl-putrescine, spermidine, spermine, N(1)-acetyl-spermine, and l-ornithine. The novelty of the work is the use of two SPE columns online coupled to LC/MS/MS, which minimizes the sample pretreatment to a single derivatization step. The analysis is complete within 4min, making the method highly suitable for routine clinical analysis and high throughput screenings. The method was fully validated with serum samples. Dynamic ranges were 0.03 to 15μg/ml for ornithine and 1 to 500ng/ml for other polyamines, which cover physiological concentrations in serum samples. Lower limits of quantification (LLoQ) were found to be between 0.1 and 5ng/ml. As a proof of concept, we investigated gender differences in polyamine levels by analyzing the serum levels of 102 subjects.
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria. Electronic address: ca.health@joanneum.at.
Christoph
C
Magnes
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria.
Alexander
A
Fauland
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria.
Edgar
E
Gander
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria.
Sophie
S
Narath
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria.
Maria
M
Ratzer
Institute of Molecular Biosciences, Karl-Franzens-University of Graz, Graz, Austria.
Tobias
T
Eisenberg
Institute of Molecular Biosciences, Karl-Franzens-University of Graz, Graz, Austria.
Frank
F
Madeo
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria; Division of Endocrinology and Metabolism, Department of Internal Medicine, Medical University of Graz, Graz, Austria.
Thomas
T
Pieber
HEALTH-Institute for Biomedicine and Health Sciences, Joanneum Research, Forschungsgesellschaft m.b.H., Graz, Austria; Division of Endocrinology and Metabolism, Department of Internal Medicine, Medical University of Graz, Graz, Austria.
Frank
F
Sinner
D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D011073:Polyamines / Q000032:analysis* / Q000097:blood; D012737:Sex Factors; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
D011073:Polyamines
Biogenic amines; Online solid phase extraction; Polyamine; Spermidine; Spermine; Tandem mass spectrometry