The Role of Fluorinated Alcohols as Mobile Phase Modifiers for LC-MS Analysis of Oligonucleotides

被引:72
作者
Basiri, Babak [1 ]
van Hattum, Hilde [2 ]
van Dongen, William D. [2 ]
Murph, Mandi M. [1 ]
Bartlett, Michael G. [1 ]
机构
[1] Univ Georgia, Coll Pharm, Dept Pharmaceut & Biomed Sci, 250 W Green St, Athens, GA 30602 USA
[2] ProQR Therapeut NV, Zernikedreef 9, NL-2333 CK Leiden, Netherlands
关键词
Oligonucleotides; LC-MS; Electrospray ionization; Fluorinated alcohols; IONIZATION MASS-SPECTROMETRY; PERFORMANCE LIQUID-CHROMATOGRAPHY; CHARGE-STATE DISTRIBUTION; ELECTROSPRAY-IONIZATION; GAS-PHASE; CONFORMATIONAL-CHANGES; G-QUADRUPLEXES; NUCLEIC-ACIDS; NEGATIVE-IONS; ESI-MS;
D O I
10.1007/s13361-016-1500-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:190 / 199
页数:10
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