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. 2018 Mar 13;8:4433. doi: 10.1038/s41598-018-22806-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Separation of nicotinamide mononucleotide (NMN) from (NAM) using uncharged chaotrope mobile phase (50 mM 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP)), isocratic elution, flow: 3 mL/min on PolyHEA column 250 × 9.4 mm, 5 μm, 60-Å. Jasco PU2089 HPLC pump. Detectors: Thermo Velos Pro linear ion trap MS running in ESI + mode, Spray Voltage: 3 kV, Capillary temp: 375 °C, H-ESI-II ion source, connected parallel to MD2015 UV-Vis detector using 1:10 post column flow splitter. Sample volume: 20 μL of standard NMN 10 mM (Sigma Aldrich). Total Ion Current (A); 260 nm absorption chromatogram (B); 335 Da ions (C); 123 Da ions (D); The same elution conditions but mobile phase supplemented with 200 mM NaCl, to eliminate the electrostatic repulsion effect, 260 nm chromatogram UV-Vis detector only (E).