Department of Pharmaceutical and Biomedical Sciences, The University of Georgia College of Pharmacy, 250 W. Green Street, Athens, GA, 30602-2352, USA.
ProQR Therapeutics NV, Zernikedreef 9, 2333 CK, Leiden, The Netherlands.
J Am Soc Mass Spectrom. 2017 Jan;28(1):190-199. doi: 10.1007/s13361-016-1500-3. Epub 2016 Sep 19.
Hexafluoroisopropanol (HFIP) has been widely used as an acidic modifier for mobile phases for liquid chromatography-mass spectrometry (LC-MS) analysis of oligonucleotides ever since the first report of its use for this purpose. This is not surprising, considering the exceptional performance of HFIP compared with carboxylic acids, which cause significant MS signal suppression in electrospray ionization. However, we have found that other fluorinated alcohols can also be utilized for mobile phase preparation and the choice of optimal fluorinated alcohol is determined by the ion-pairing (IP) agent. Although HFIP is a very good choice to be used alongside less hydrophobic IP agents, other fluorinated alcohols such as 1,1,1,3,3,3-hexafluoro-2-methyl-2-propanol (HFMIP) can significantly outperform HFIP when used with more hydrophobic IP agents. We also found that more acidic fluorinated alcohols assist with the transfer of oligonucleotides with secondary structure (e.g., folded strands and hairpins) into the gas phase. Graphical Abstract ᅟ.
六氟异丙醇 (HFIP) 自首次被报道用于寡核苷酸的液相色谱-质谱 (LC-MS) 分析的流动相酸性改性剂以来，已被广泛应用。这并不奇怪，因为与羧酸相比，HFIP 的性能非常出色，羧酸在电喷雾电离中会导致显著的 MS 信号抑制。然而，我们发现其他氟化醇也可用于流动相制备，而最佳氟化醇的选择取决于离子对试剂 (IP)。尽管 HFIP 是与疏水性较低的 IP 试剂一起使用的绝佳选择，但在与疏水性更高的 IP 试剂一起使用时，其他氟化醇，如 1,1,1,3,3,3-六氟-2-甲基-2-丙醇 (HFMIP)，性能可显著优于 HFIP。我们还发现，酸性更强的氟化醇有助于将具有二级结构（例如折叠链和发夹）的寡核苷酸转移到气相中。
