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Similarly, substrate nicotinamide at 100 μM, methyl donor S-adenosyl-L-methionine (SAM) at 5 μM, and S-adenosyl methionine (SAH) at 5 μM concentration) samples were also run individually in the reaction buffer [1 mM Tris [pH 8.6], 1 mM DTT, 10% trichloroacetic acid, 4% methanol, and water] to identify elution time and define substrate, co-factor, and product peaks