High-speed gradient elution reversed-phase liquid chromatography of bases in buffered eluents

High speed gradient elution reversed-phase liquid chromatography.

A major disadvantage of gradient elution in terms of speed results from the need to adequately re-equilibrate the column. This work distinguishes two states of re-equilibration: (1) run-to-run repeatability and (2) full equilibration. We find that excellent repeatability (+/-0.002 min in retention time) is achieved with at most 2 column volumes of re-equilibration whereas full equilibration can...

New approach to linear gradient elution used for optimisation in reversed-phase liquid chromatography.

A new mathematical treatment concerning the gradient elution in reversed-phase liquid chromatography when the volume fraction psi of an organic modifier in the water-organic mobile phase varies linearly with time is presented. The experimental ln k versus psi curve, where k is the retention factor under isocratic conditions in a binary mobile phase, is subdivided into a finite number of linear ...

Reversed-phase high-performance liquid chromatography.

Solubility of Buffers in Aqueous–Organic Eluents for Reversed-Phase Liquid Chromatography

Address correspondence to P.W. Carr. hen preparing binary eluents containing both an organic solvent and a buffer for a buffer reversed-phase liquid chromatography (LC), one must know the solubility of the buffer in the organic cosolvent to avoid precipitation of the buffer salt. For example, one must know the highest percent organic cosolvent (that is, %B, v/v) in which a buffer salt remains s...

Analysis of sphingoid bases by reversed-phase high performance liquid chromatography.

A reversed-phase high performance liquid chromatography (HPLC) technique was developed for the analysis of sphingoid bases as their biphenylcarbonyl derivatives. The sphingoid bases (0.3-500 nmol) obtained from sphingolipids after hydrolysis were quantitatively derivatized with biphenylcarbonyl chloride at room temperature for 90 min. After removal of excess reagent by partitioning, the derivat...