Purification of commercial preparations of NADP+ from AMP contamination.
A method for purification of commercial preparations of NADP+ from AMP contamination is described. The purification procedure includes one-step anion-exchange chromatography on Dowex-1 (formate) and results in a highly purified salt-free coenzyme with a yield of 70-80%. The chromatography conditions have been selected allowing for complete separation of AMP from NADP+ in a HCOOH concentration gradient. This is followed by NADP+ elution with 1.5 M HCOOH containing HCOOK at a concentration at which the salt remains in solution during subsequent precipitation and washing of NADP+ with acetone. An addition of HCOOK is necessary to reduce the coenzyme elution volume that is important for further precipitation of NADP+ with acetone.
D000249:Adenosine Monophosphate; D000818:Animals; D002852:Chromatography, Ion Exchange; D004340:Drug Contamination; D009249:NADP / Q000302:isolation & purification*; D011817:Rabbits
D000249:Adenosine Monophosphate; D009249:NADP