Application note:
Frédérique L. van Holthoon, William D. van Dongen, Anne J. Kleinnijenhuis
Triskelion, Zeist, The Netherlands
For clinical development of biopharmaceuticals it is required to investigate their in vivo behavior using validated bioanalytical methods In guidances for bioanalytical method validation 1 3 it is strongly recommended to investigate matrix effects for the analyte(s) in hemolyzed plasma or serum, in addition to clean matrix During hemolysis, the contents of red blood cells, mainly hemoglobin, will be released into the surrounding matrix When UPLC MS (ultra performance liquid chromatography mass spectrometry) is applied, hemolysis can affect analyte suppression or enhancement and the
extraction efficiency 4 or analyte stability.
