Volatile Biomarkers in Breath Associated With Liver Cirrhosis — Comparisons of Pre- and Post-liver Transplant Breath Samples

• Breath volatiles were compared for cirrhotic patients and controls and pre- and post-liver transplant.
• Three volatiles (limonene, methanol, 2-pentanone) have been found to have excellent diagnostic capabilities.
• Limonene shows washout characteristics following transplant supporting a hypothesis that it accumulates in fat.There are numerous previous studies investigating breath volatiles in patients with liver disease but with conflicting results. It is impossible to tell which volatiles from previous studies may be false discoveries, and which are actually associated with the disease. We measured breath samples in patients and controls and in patients after transplant. Methanol, 2-pentanone and limonene show differences not only between patients and controls but also in cases pre- and post-transplant and have excellent diagnostic capabilities. We show evidence that limonene accumulates in the body, probably because the cirrhotic liver fails to metabolise dietary limonene.

BackgroundThe burden of liver disease in the UK has risen dramatically and there is a need for improved diagnostics.AimsTo determine which breath volatiles are associated with the cirrhotic liver and hence diagnostically useful.MethodsA two-stage biomarker discovery procedure was used. Alveolar breath samples of 31 patients with cirrhosis and 30 healthy controls were mass spectrometrically analysed and compared (stage 1). 12 of these patients had their breath analysed after liver transplant (stage 2). Five patients were followed longitudinally as in-patients in the post-transplant period.ResultsSeven volatiles were elevated in the breath of patients versus controls. Of these, five showed statistically significant decrease post-transplant: limonene, methanol, 2-pentanone, 2-butanone and carbon disulfide. On an individual basis limonene has the best diagnostic capability (the area under a receiver operating characteristic curve (AUROC) is 0.91), but this is improved by combining methanol, 2-pentanone and limonene (AUROC curve 0.95). Following transplant, limonene shows wash-out characteristics.ConclusionsLimonene, methanol and 2-pentanone are breath markers for a cirrhotic liver. This study raises the potential to investigate these volatiles as markers for early-stage liver disease. By monitoring the wash-out of limonene following transplant, graft liver function can be non-invasively assessed.