Cells were incubated in 2 ml culture medium with 10 μM 2-ClHDyA for 5 h. At indicated time points, medium and cells were extracted in the presence of the internal standards 2-ClHA and 2-ClHOH (100 ng each). After conversion to the corresponding PFB-ester derivatives the content of 2-ClHyA and 2-ClHyOH of (A) cells and (B) the cellular supernatant was quantitated by NICI-GC-MS analysis. Results are displayed as mean values ± SD of triplicate determinations.
Data in (C) represent loss of 2-ClHDyA from the medium (Fig. 3A) versus recovery of 2-ClHyX (sum of 2-ClHDyA, 2-ClHyA, and 2-ClHDyOH) in cells and the cellular supernatant (from Figs. 3B, 4A and B).