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Half-lives ( t 1/2 ) in respective buffers and half-lives for GSH adduct formation ( t 1/2 GSH) were calculated with the following equations: Mass spectrometry analysis of PDE adducts after Glu-C digestion For each replicate, 1.5 g covalent adduct of PDE sample in PBS buffer was denatured, reduced, alkylated and digested by Glu-C and desalted, together with a negative control of unbound vehicle-treated PDE sample prior to mass spectrometry analysis