Figure 16.
HPLC elution profiles of MAT time-dependent reactions with (A) TFSeM (as 7b) or (B) SeM as substrates. Authentic standard compounds are shown in the black dotted traces. Elution times in minutes are as follows: 3.7 (ATP); 4.2 (ADP); 7.8 (AMP); 11.5 (SAM); 9.8 and 12.3 (SeSAM); 18.2 (adenine); 21.8 (SAH); 25.2 (tryptophan IS); 29.7 (MTA); 31.3 (SeMTA). TFSeM resulted in no apparent formation of TFSeSAM or SeSAM over the course of 60 min, whereas SeM was converted to SeSAM by MAT as evidenced by the time-dependent formation of SeSAM (elution times of 9.8 and 12.3 min). The two elution times for SeSAM match that of the authentic standard; under Method 1 conditions SeSAM elutes as one sharp, defined peak, indicating the double peak is a result of the chromatographic conditions.