The observed change in ADC DAR post-dose enables evaluation of ADC deconjugation over time. The DAR can be calculated by comparing the total Ab and ADC data from the absolute LC-MRM quantification (Figure ), or with the identified LC and HC species from relative intact LC-HRMS (Figures ). It is notable that both methods, to various extent, showed that ADC1 and 4 had a slower deconjugation rate over the time compared to ADC2 and 3. However, LC-HRMS assay was able to also characterize the structural differences in various species and kinetics of associated reactions.