Featuring ultimate sensitivity of high-resolution LC-MS analysis of phenolics in rat plasma.

Sensitive analysis of very low molecular weight metabolites using LC with quadrupole-time-of-flight MS is challenging due to the high losses of ions in a time-of-flight analyzer. Improvement in sensitivity for these analytes via the optimization of advanced parameters, including quadrupole profile, ion guide parameters, and duty cycle, has been achieved. The optimization of the method was carried out using a large spectrum of structurally different compounds, including (iso)flavonoids and their known metabolites. These compounds can be categorized into two major groups, i.e., compounds with (iso)flavonoid core and low molecular weight phenolics. The optimization of the duty cycle enabled up to a 15-fold increase in analyte responses while the contribution of tuning ion optic and quadrupole profile was negligible. The limits of quantifications of our new method were assessed using both standard solutions and rat plasma. They were decreased at least ten times for several low molecular weight phenolics enabling measurement of their concentrations in a range of 1-50 ng/mL in rat plasma after protein precipitation. Concurrently, the limits of quantifications for compounds with (iso)flavonoid core did not increase distinctly allowing their detection in a range of 0.5-10 ng/mL. The new method was used for the targeting of phenolics in biological samples from pharmacokinetics experiments. This article is protected by copyright. All rights reserved.