FIG. 5.
NADP(H) extraction from mouse liver. (A) Data show metabolite yields using seven different extraction methods. (B) Repeated extractions with 40:40:20 with 0.1 M formic acid. Two μM of 13C1015N5-ADP was added to the extraction solvent to track overall metabolite recovery. The relative amount of 13C1015N5-ADP, NADPH, and NADP+ detected in 1st, 2nd, and 3rd extracts was determined. Y-axis refers to the labeled ADP standard. For NADPH and NADP+, we assume that yield in the 1st extract is equivalent to that of the ADP standard, and amounts in subsequent extracts are relative to the 1st extract. Dash line shows signal for an equivalent amount of 13C1015N5-ADP standard. Results are the average of three replicates and the error bar is the SD. ADP, adenosine diphosphate