Lochmüller C H, Liu Q, Huang L, Li Y
Gross Chemical Laboratory, Duke University, Durham, NC 27708, USA.
J Chromatogr Sci. 1999 Jul;37(7):251-4. doi: 10.1093/chromsci/37.7.251.
This paper is the first report on the retention behavior of synthetic oligonucleotides and nucleotide oligomers on a continuous-bedmatrix, strong-anion-exchange column. The separation mechanism is predominantly an anion-exchange process, but hydrophobic interaction plays a role as well. The separation is based on the chain length of the oligonucleotide. Both the addition of organic mobile phase modifiers and changes in column temperature affect the retention of oligomers significantly. A volatile buffer system (e.g., triethylamine acetate) could be employed to purify oligonucleotides, and no desalting procedure would be required after the column separation step. The recoveries from the separation are 70% or higher. The maximum loading capacity of an analytical column (35 x 7-mm i.d.) was found to be more than 366 micrograms.
本文是关于合成寡核苷酸和核苷酸低聚物在连续床基质强阴离子交换柱上保留行为的首次报道。分离机制主要是阴离子交换过程，但疏水相互作用也起作用。分离基于寡核苷酸的链长。有机流动相改性剂的添加和柱温的变化均会显著影响低聚物的保留。可采用挥发性缓冲体系（如三乙胺乙酸盐）来纯化寡核苷酸，柱分离步骤后无需脱盐程序。分离回收率达70%或更高。发现一根分析柱（内径35×7 mm）的最大进样量超过366微克。
