Validation of an LC-MS/MS Method for Urinary Lactulose and Mannitol Quantification: Results in Patients with Irritable Bowel Syndrome

Franco Scaldaferri, Andrea Urbani, Silvia Persichilli, Jacopo Gervasoni, Arcangelo Schiattarella, Valentina Giorgio, Aniello Primiano, Valentina Tesori, Cecilia Zuppi

Risultato della ricerca: Contributo in rivistaArticolo in rivista

8 Citazioni (Scopus)

Abstract

Aim. Lactulose/mannitol ratio is used to assess intestinal barrier function. Aim of this work was to develop a robust and rapid method for the analysis of lactulose and mannitol in urine by liquid chromatography coupled to tandem mass spectrometry. Lactulose/mannitol ratio has been measured in pediatric patients suffering from irritable bowel syndrome. Methods. Calibration curves and raffinose, used as internal standard, were prepared in water : acetonitrile 20:80. Fifty mu L of urine sample was added to 450 mu L of internal standard solution. The chromatographic separation was performed using a Luna NH2 column operating at a flow rate of 200 mu L/min and eluted with a linear gradient from 20% to 80% water in acetonitrile. Total run time is 9 minutes. The mass spectrometry operates in electrospray negative mode. Method was fully validated according to European Medicine Agency guidelines. Results and Conclusions. Linearity ranged from 10 to 1000 mg/L for mannitol and 2.5 to 1000 mg/L for lactulose. Imprecision in intra-and interassay was lower than 15% for both analytes. Accuracy was higher than 85%. Lactulose/mannitol ratio in pediatric patients is significantly higher than that measured in controls. The presented method, rapid and sensitive, is suitable in a clinical laboratory.
Lingua originaleEnglish
pagine (da-a)1-6
Numero di pagine6
RivistaDisease Markers
Volume2016
DOI
Stato di pubblicazionePubblicato - 2016

Keywords

  • AUTOMATED ENZYMATIC ASSAYS
  • INTESTINAL PERMEABILITY PROBES

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