A LC Infinity II 1290 (Agilent) coupled to a Zeno TOF 7600 with a Turbo V source (SCIEX) was used. Chromatographic separation was based on the same method described by Jamnik et al (2022). In summary, a 20 min reversed-phase gradient was employed on an HSS T3 column (Waters), with a flow rate of 0.4 mL/min and an injection volume of five microliters. LC-MS data acquisition was performed for both a SWATH-only method and the newly developed Zeno MRM-HR + SWATH methods. Ion source parameters were standardized across both methodologies, as follow: GS1 -50 psi; GS2 -50 psi; curtain gas -35 psi; CAD gas -9; temperature -550 °C. In both cases, MS 1 data was acquired from m/z 100 to 1000, with declustering potential at 80V (-80 V for negative mode) and collision energy at 10V (-10 V for negative mode). Spray voltage was set at 5500 V for positive ionization mode (ESI+) and -4500 V for negative ionization mode (ESI-). For the developed Zeno MRM-HR + SWATH data, accumulation time was set at 0.05 s for individuals MRM transitions and at 0.02 s for each SWATH window (n= 10). SWATH windows were optimized individually for each SRM using the SWATH windows optimizer provided by SCIEX. Both SWATH-only and MRM-HR+SWATH methods had the exact same parameter for SWATH acquisition, including windows range, declustering potential (80 or -80V) and CE, as described in details in Table . For MRM experiment, Start/Stop option was selected, with TOF Start Mass at m/z 50 and TOF Stop Mass set as compound-dependent by adding 2 m/z units to precursor m/z. Retention time windows, declustering potentials and collision energies were adjusted for each compound (see Results section) and Zeno Pulsing was activated at a threshold of 8000 cps. To allow for a point of comparison, the cycle time for the SWATH-only method was adjusted to a similar value as the maximum scan time for the MRM-HR + SWATH method. To evaluate possible differences in increased accumulation time for MS or SWATH windows, two different approaches were taken. For the Zeno MRM-HR+SWATH in negative mode, accumulation time for MS was increased from 0.05 s to 0.15 s (for both SRM 1958 and SRM 1950). For positive mode, on the other hand, MS 1 accumulation time was kept the same as for MRM-HR + SWATH method (0.05 s) and, instead, accumulation times for SWATH-only windows were increased such as total cycle time would compare to maximum cycle time for MRM-HR + SWATH, as previously explained. For instance, maximum scan time for Zeno MRM-HR +SWATH method was 0.435 s while SWATHonly total cycle time was 0.25 s (0.05 s for MS1 and 0.02 s for each of the 10 SWATH window). Therefore, at the maximum number of overlapping MRM windows, the total cycle time would be 0.685 s. Based on that, the SWATH-only method for positive mode was set as total scan time of 0.643 by keeping MS1 acquisition at 0.05 s and increasing SWATH windows accumulation time from 0.02 to 0.05 s. For direct comparison, key method parameters are also summarized in Table .