Enzymatic treatment efficiently removes ADPR and ADPRP contamination of pyridine dinucleotides. (A–C) TLC images showing the purification of (A) NAD, (B) NAAD, and (C) NAADP. Loaded samples (1 µl) were as follows: (A) 10 mM ADPR (lane 1), 10 mM AMP (lane 2), 10 mM NAD (lane 3), and 100 mM NAD before (lane 4) and after (lane 5) treatment with NUDT9. (B) 10 mM ADPR (lane 1), 10 mM AMP (lane 2), 10 mM NAAD (lane 3), and 100 mM NAAD before (lane 4) and after (lane 5) treatment with NUDT9. (C) 10 mM NAADP (lane 1), 10 mM ADPRP (lane 2), 10 mM AMPP (lane 3), and 100 mM NAADP before (lane 4) and after (lane 5) treatment with NUDT9. (D) Hydrolytic activity of NUDT9 on ADPR and ADPRP, showing 10 mM ADPR (lane 1), 10 mM AMP (lane 2), 10 mM ADPR treated with NUDT9 (lane 3), 10 mM ADPRP (lane 4), 10 mM AMPP (lane 5), 10 mM ADPRP treated with NUDT9 (lane 6), mixture of 10 mM ADPR and 10 mM ADPRP before (lane 7), and after (lane 8) treatment with NUDT9. Developing solution was DS2 in A and C, and DS1 in B and D.