A non-invasive breath test for research to measure hepatic metabolic capacity

LIBRA CASE STUDy examples

Comparing Healthy and Cirrhotic Subjects

Using the LIBRA Test, limonene breath profiles can be generated for each participant. As seen in figure 3, limonene ingestion elicits a >100-fold spike of breath limonene compared to baseline. The investigated timepoints post-administration showed excellent classification performance. At 60 minutes we measured an area under the roc curve (AUROC) of 0.91, sensitivity of 0.83 ± 0.07, and specificity of 0.9 ± 0.06 (Ferrandino et al. Biomedicine 2023).

Ferrandino and colleagues effectively identified subjects who had been incorrectly allocated into the cirrhotic and healthy control groups. Figure 4 demonstrates that subject DYL10297 was initially recruited as a control but exhibited an abnormal breath profile. A follow-up ultrasound revealed previously unknown liver disease. Similarly, Subject DYL10008, classified in the cirrhosis group based on a three-year-old autoimmune hepatitis diagnosis, displayed a breath profile resembling that of healthy subjects. A further ultrasound confirmed cirrhosis regression, suggesting successful immunosuppressant treatment. These findings highlight LIBRA’s potential for both chronic liver disease detection and treatment monitoring.

Detecting Cirrhosis in a High Risk Population

In a recent unpublished study, LIBRA identified subjects with cirrhosis. The cohort included high-risk individuals who had symptoms of cirrhosis. 78 of 147 individuals were diagnosed with early compensated cirrhosis across Chile, the United Kingdom, and the United States. Figure 5 shows the classification performance, with an area under the curve (AUC) of 0.82, sensitivity of 0.73 and specificity of 0.73 (at Youden index).

Figure 6a demonstrates that a progressive increase of EVOCs on breath was observed across the spectrum of liver fibrosis estimated using FibroScan®. In figure 6b, we can see breath limonene 15 minutes post administration of the LIBRA Oral Solution. There is a positive correlation between limonene levels and FIB-4 in subjects with cirrhosis.

Portal Hypertension

Dietary limonene is has been associated with signs of portal hypertension. Figure 7 a) is an analysis of 133 cirrhotic patients across four studies and revealed higher breath limonene levels derived from dietary exposure in individuals with thrombocytopenia (platelet count <150 × 10/L).

In an exploratory study of 29 cirrhotic patients, ultrasound evaluation of spleen size showed that those with splenomegaly (spleen >12 cm) exhibited increased breath limonene levels from random dietary exposure (figure 7 b).

Since thrombocytopenia and splenomegaly are established markers of portal hypertension, these findings suggest LIBRA’s potential for longitudinal monitoring, aiding in the early detection of portal hypertension and timely intervention.

Chronic Liver Disease

A progressive rise in limonene through the chronic liver disease spectrum has been seen in four independent studies conducted by Owlstone Medical. These looked at limonene levels found on breath from dietary intake (no intervention). Figure 8 illustrates that healthy individuals display low levels of breath limonene, whilst pre-cirrhotic and cirrhotic patients show increasingly high levels. Changes in VOCs are thus detectable at pre-cirrhotic stages, suggesting early biomarkers for liver dysfunction.

The LIBRA test may facilitate early identification of individuals with chronic liver disease before the cirrhosis stage, enabling early intervention. The progressive nature of VOC alterations could serve for monitoring liver function improvement.

Would you be interested in using the LIBRA test in your research? Do not hesitate to get in touch to learn more about how we can help.