Oxidative stress is a pathological condition characterized by an imbalance between body's antioxidant defenses and oxidizing agents, resulting in damage of endogenous molecules. These products can be used as markers of oxidative conditions; in particular, isoprostanes (IsoPs) come from the reaction of arachidonic acid with reactive oxygen species (ROS) and are currently defined as gold markers of oxidative stress in urine. Our main goal was the development of a reliable analytical method for the determination and quantification of the IsoPs in human urine by dispersive Liquid–Liquid Micro Extraction (dLLME) coupled with micro Solid Phase Extraction (μSPE) clean-up and HPLC-MS/MS analysis. The selected compounds are present in very small concentration in urine, furthermore, due to relevant matrix effect, they are challenging for ESI-MS/MS analysis. This approach provided selectivity and sensitivity for 8-isoprotaglandine F2α (8-iso-PGF2α), the “gold” OS marker, together with the main isomers. dLLME extraction allowed a significant enrichment factor and μSPE clean-up provided the removal of ion-suppressing compounds from the sample resulting in low matrix effect. The chromatographic separation was also challenging as the target compounds possess very similar chemical characteristics, so experimental conditions were carefully tuned. The reported method represents a useful tool for the detection of IsoPs in urine taking advantage of the combination of dLLME extraction and μSPE clean-up; overall recoveries were above 50 % and matrix effects were ≤15 %, with LOQs ranging between 0.020 and 0.060 ng mL−1. The procedure is easy to use and rapid allowing the removal of interfering compounds and matrix effect maintaining a highly sensitive determination.

dLLME-μSPE extraction coupled to HPLC-ESI-MS/MS for the determination of F2α-IsoPs in human urine / Fanti, F.; Vincenti, F.; Montesano, C.; Serafini, M.; Compagnone, D.; Sergi, M.. - In: JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS. - ISSN 0731-7085. - 186:(2020), pp. 1-7. [10.1016/j.jpba.2020.113302]

dLLME-μSPE extraction coupled to HPLC-ESI-MS/MS for the determination of F2α-IsoPs in human urine

Vincenti F.;Montesano C.;Compagnone D.;Sergi M.
2020

Abstract

Oxidative stress is a pathological condition characterized by an imbalance between body's antioxidant defenses and oxidizing agents, resulting in damage of endogenous molecules. These products can be used as markers of oxidative conditions; in particular, isoprostanes (IsoPs) come from the reaction of arachidonic acid with reactive oxygen species (ROS) and are currently defined as gold markers of oxidative stress in urine. Our main goal was the development of a reliable analytical method for the determination and quantification of the IsoPs in human urine by dispersive Liquid–Liquid Micro Extraction (dLLME) coupled with micro Solid Phase Extraction (μSPE) clean-up and HPLC-MS/MS analysis. The selected compounds are present in very small concentration in urine, furthermore, due to relevant matrix effect, they are challenging for ESI-MS/MS analysis. This approach provided selectivity and sensitivity for 8-isoprotaglandine F2α (8-iso-PGF2α), the “gold” OS marker, together with the main isomers. dLLME extraction allowed a significant enrichment factor and μSPE clean-up provided the removal of ion-suppressing compounds from the sample resulting in low matrix effect. The chromatographic separation was also challenging as the target compounds possess very similar chemical characteristics, so experimental conditions were carefully tuned. The reported method represents a useful tool for the detection of IsoPs in urine taking advantage of the combination of dLLME extraction and μSPE clean-up; overall recoveries were above 50 % and matrix effects were ≤15 %, with LOQs ranging between 0.020 and 0.060 ng mL−1. The procedure is easy to use and rapid allowing the removal of interfering compounds and matrix effect maintaining a highly sensitive determination.
2020
dispersive liquid liquid micro extraction; HPLC-MS/MS; isoprostanes; micro SPE; urine
01 Pubblicazione su rivista::01a Articolo in rivista
dLLME-μSPE extraction coupled to HPLC-ESI-MS/MS for the determination of F2α-IsoPs in human urine / Fanti, F.; Vincenti, F.; Montesano, C.; Serafini, M.; Compagnone, D.; Sergi, M.. - In: JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS. - ISSN 0731-7085. - 186:(2020), pp. 1-7. [10.1016/j.jpba.2020.113302]
File allegati a questo prodotto
File Dimensione Formato  
Fanti_dLLME_2020.pdf

solo gestori archivio

Tipologia: Documento in Post-print (versione successiva alla peer review e accettata per la pubblicazione)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 770.63 kB
Formato Adobe PDF
770.63 kB Adobe PDF   Contatta l'autore

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1411295
Citazioni
  • ???jsp.display-item.citation.pmc??? 0
  • Scopus 9
  • ???jsp.display-item.citation.isi??? 7
social impact