Rapid quantification of bile acids in serum by LC-MS/MS and application to serum bile acid profile in voriconazole administered patients with invasive fungal infections

Disruption of bile acid homeostasis is implicated in the pathogenesis and progress of several liver diseases, including drug-induced liver injury (DILI). A rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying 30 bile acids in human serum was developed and applied to analyze serum bile acid profile in voriconazole administered patients with invasive fungal infections. The stable isotope-coded bile acids were used as internal standards. The analytes in serum samples (50 μL) were extracted by protein precipitation and isolated by a Kinetex EVO C18 column (50 × 2.1 mm, 2.6 μm). The detection was performed using multiple reaction monitoring (MRM) in electrospray negative ionization mode. All of the 30 bile acids were sufficiently separated within 10 min with good linearity (r > 0.99) over tested calibration ranges. The accuracy and precision values were in the range of 85.42–114.3% and 1.8–13.8%, respectively. The matrix effect, recovery, stability, and dilution integrity all met the acceptance criteria. Serum bile acid profile changed in patients with supratherapeutic voriconazole concentration (>10 μg/mL). This LC-MS/MS method for the quantification of 30 bile acids in human serum might apply a new analytical approach for liver function evaluation. We developed an easy, fast, and reliable method to measure 30 bile acids in human serum, using a technique called LC-MS/MS. Bile acids are substances produced in the body that help with energy regulation, and they also act as chemical messengers in the body. When the balance of bile acids is disturbed, it may play a role in the development and worsening of some liver diseases, including liver damage caused by certain medications. For this reason, having a trustworthy way to measure bile acids is important. Our new method was highly accurate and consistent, and it could measure 30 bile acids in a serum sample at the same time. The bile acids we measured included several main types, as well as their conjugated forms that were combined with two small molecules called glycine or taurine. This method was well-suited for use in clinic because it was simple to perform and could process many samples quickly. We used it to evaluate the levels of bile acids in patients being treated for invasive fungal infections with a drug called voriconazole. The results showed that patients who had higher-than-recommended levels of voriconazole in their blood (more than 10 μg/mL) had different bile acid levels compared to normal conditions. These findings helped us better understand the relationship between voriconazole-induced liver damage and bile acid levels, and could support further research into how this drug impacts the liver.