Ion-pair reversed-phase high-performance liquid chromatography analysis of oligonucleotides: retention prediction

An ion-pair reversed-phase HPLC method was evaluated for the separation of synthetic oligonucleotides. Mass transfer in the stationary phase was found to be a major factor contributing to peak broadening on porous C18 stationary phases. A small sorbent particle size (2.5 microm), elevated temperature and a relatively slow flow-rate were utilized to enhance mass transfer. A short 50 mm column allows for an efficient separation up to 30mer oligonucleotides. The separation strategy consists of a shallow linear gradient of organic modifier, optimal initial gradient strength, and the use of an ion-pairing buffer. The triethylammonium acetate ion-pairing mobile phases have been traditionally used for oligonucleotide separations with good result. However, the oligonucleotide retention is affected by its nucleotide composition. We developed a mathematical model for the prediction of oligonucleotide retention from sequence and length. We used the model successfully to select the optimal initial gradient strength for fast HPLC purification of synthetic oligonucleotides. We also utilized ion-pairing mobile phases comprised of triethylamine (TEA) buffered by hexafluoroisopropanol (HFIP). The TEA-HFIP aqueous buffers are useful for a highly efficient and less sequence-dependent separation of heterooligonucleotides.     

Idempotent bounded C-semigroups

A semigroup with zero isidempotent bounded (IB) if it is the 0-direct union of idempotent generated principal left ideals and the 0-direct union of idempotent generated principal right ideals. Notable examples are completely 0-simple semigroups and the wider class of primitive abundant semigroups. Significant to the structure of these semigroups is that they are all categorical at zero. In this paper we describe IB semigroups that are categorical at zero in terms ofdouble blocked Rees matrix semigroups. This generalises Fountain's characterisation of primitive abundant semigroups via blocked Rees matrix semigroups [1], which in turn yields the Rees theorem for completely 0-simple semigroups.