Entity: Universitat Rovira i Virgili (URV)
Confidenciality: No
Education area(s): Genètica, Física i Química Forense
Title in different languages: Determination of polyamines in urine by UHPLC-MS/MS
Abstract: In the last years, it has been shown that polyamines are a potential candidate to act as cancer biomarkers. In this work a method has been developed by coupling ultra-high performance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of polyamines precursors, polyamines and polyamine catabolite in human urine. Polyamine precursors including ornithine, lysine and arginine; polyamines including agmatine, putrescine, cadaverine, 1,3-diaminopropane, spermidine, spermina, N-acetylputrescine, N1-acetylspermidine, N8-acetylspermidine, N-acetylspermine, N1,N8-diacetylspermidine and N1,N12-dialcetylspermine; polyamine catabolite including ϒ-aminobutyric acid had been determined. The samples have been pre-treated using different procedures to compare them and choose the most suitable. The first step consists of a precipitation of the polyamines by methanol and chloroform or percloric acid extraction, followed by derivatization with dansyl chloride. The last extraction has been carried out with solid phase extraction or liquid-liquid extraction with ethyl acetate and TFA. The procedure selected has been extraction with methanol and chloroform/ derivatization/ Liquid-Liquid due to its lower matrix effect and better recovery. Derived polyamines have been separated with a C18 column using a gradient program. The detection has been performed with the UHPLC-MS/MS system with electrospray ionitzation source in the positive ion (ESI+) and multiple reactions-monitoring mode (MRM). The calibration curve has been obtained for each polyamine with a determination coefficient greater than 0.993. The limits of quantitation for all analytes were within 0.04 and 6.86 ng/ml. The limits of quantitation for all analytes were within 4 amb 32000 ng/ml in urine. Finally, the intra-and inter-day CVs for all analyts were 5.23-21.66% and 0.78-18.24%, respectively.
Subject: Química
Academic year: 2017-2018
Language: Català
Work's public defense date: 2018-06-26
Subject areas: Chemistry
Student: Ribas López, Verónica
Work's codirector: Del Pino, Antoni Rius
Department: Química Analítica i Química Orgànica
Creation date in repository: 2018-09-26
TFM credits: 18
Keywords: Determination, polyamines, urine
Title in original language: Determinació de poliamines en orina mitjançant UHPLC-MS/MS
Access Rights: info:eu-repo/semantics/openAccess
Project director: Aguilar Anguera,Carme
Repositori URV