Premium
Retention of nucleic acids in ion‐pair reversed‐phase high‐performance liquid chromatography depends not only on base composition but also on base sequence
Author(s) -
Qiao Junqin,
Liang Chao,
Wei Lanchun,
Cao Zhaoming,
Lian Hongzhen
Publication year - 2016
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201600701
Subject(s) - nucleic acid , oligonucleotide , chemistry , chromatography , guanine , composition (language) , base pair , hydrophilic interaction chromatography , phase (matter) , high performance liquid chromatography , cytosine , base (topology) , reversed phase chromatography , memory retention , dna , nucleotide , biochemistry , organic chemistry , biology , gene , mathematical analysis , linguistics , philosophy , mathematics , neuroscience
The study on nucleic acid retention in ion‐pair reversed‐phase high‐performance liquid chromatography mainly focuses on size‐dependence, however, other factors influencing retention behaviors have not been comprehensively clarified up to date. In this present work, the retention behaviors of oligonucleotides and double‐stranded DNAs were investigated on silica‐based C 18 stationary phase by ion‐pair reversed‐phase high‐performance liquid chromatography. It is found that the retention of oligonucleotides was influenced by base composition and base sequence as well as size, and oligonucleotides prone to self‐dimerization have weaker retention than those not prone to self‐dimerization but with the same base composition. However, homo‐oligonucleotides are suitable for the size‐dependent separation as a special case of oligonucleotides. For double‐stranded DNAs, the retention is also influenced by base composition and base sequence, as well as size. This may be attributed to the interaction of exposed bases in major or minor grooves with the hydrophobic alky chains of stationary phase. In addition, no specific influence of guanine and cytosine content was confirmed on retention of double‐stranded DNAs. Notably, the space effect resulted from the stereostructure of nucleic acids also influences the retention behavior in ion‐pair reversed‐phase high‐performance liquid chromatography.