Determination of putrescine, cadaverine, spermidine and spermine in different chemical matrices by high performance liquid chromatography-electrospray ionization-ion trap tandem mass spectrometry (HPLC-ESI-ITMS/MS).
The goal of this work was to establish a simple HPLC-ESI-ITMS/MS procedure, suitable for the determination of four common aliphatic polyamines in two different types of biological matrices. To this end, 1,6-diaminohexane was used as the internal standard (IS) and 4-fluoro-3-nitrobenzenotrifluoride (FNBT) as the derivatizing agent. Formation of fully derivatized compounds was confirmed by high resolution ESI-QTOFMS and MS/MS analysis. Reversed phase chromatographic separation was carried out by gradient elution with 0.1% (v/v) formic acid and methanol. In a positive ESI mode, the following pairs of precursor/quantifier ions were used for multiple reaction monitoring: 467.4/261.0 for PUT, 481.2/461.1 for CAD, 713.7/261.0 for SPD, 959.8/507.2 for SPM and 495.3/475.2 for IS. On-column instrumental detection limits of four polyamines were in the range 0.62-2.14fmol (0.039-0.215ng/ml). Versatility was demonstrated by analyzing plant extracts and human urine; prior to derivatization, all samples were cleaned-up by dichloromethane extraction. The evaluated signal suppression/enhancement was in the range 82.3-115.4% and the percentage recoveries obtained in the method of standard addition were in the range 83.7-114.4%. Statistically significant differences in polyamines concentrations were found in garden cress exposed to Cd(II) versus control seedlings (t-test, p<0.05); results obtained for urine from healthy volunteers and diabetic patients at different clinical conditions suggested possible utility of free polyamines as biomarkers of progressive diabetes.
Department of Chemistry, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico;
Alan Alexander González
AA
Ibarra
Department of Chemistry, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico;
Katarzyna
K
Wrobel
Department of Chemistry, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico;
Alma Rosa Corrales
AR
Escobosa
Department of Chemistry, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico;
Julio Cesar Torres
JC
Elguera
Department of Medical Sciences, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico.
Ma Eugenia
ME
Garay-Sevilla
Department of Chemistry, University of Guanajuato, L. de Retana 5, 36000 Guanajuato, Mexico;. Electronic address: kazimier@ugto.mx.
Kazimierz
K
Wrobel
D015306:Biogenic Monoamines / Q000032:analysis* / Q000652:urine; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003920:Diabetes Mellitus / Q000652:urine; D006801:Humans; D057230:Limit of Detection; D010936:Plant Extracts / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D015306:Biogenic Monoamines; D010936:Plant Extracts
4-fluoro-3-nitrobenzenotrifluoride (FNBT); Aliphatic polyamines; Diabetes; LC–MS, Multiple reaction monitoring; Plant abiotic stress; Urine