Use of Dyes To Investigate Migration of the Chiral Selector in CFFE and the Impact on the Chiral Separations

Abstract

Continuous free flow electrophoresis was investigated as a tool for the preparative chiral separation of piperoxan enantiomers using sulfated beta-cyclodextrin (sbeta-CD) as the chiral additive. Bulk migration of sbeta-CD was confirmed using LC-MS analysis of the individual fractions collected and visualized with the addition of crystal violet to the separation buffer. In the absence of sbeta-CD, the crystal violet-containing buffer was reddish/purple and the crystal violet was deflected cathodically in the chamber. In the presence of sbeta-CD, the crystal violet-containing buffer was blue and was deflected anodically. However, formation of accumulation and depletion zones was apparent in both cases. The addition of sbeta-CD to the cathodic wash solution allowed for almost complete resolution of the piperoxan enantiomers with a processing rate of 0.45 mg/ h.

Quantitative investigation of resolution increase of free‐flow electrophoresis via simple interval sample injection and separation

“…As an important preparative purification technique , free‐flow electrophoresis (FFE) has been used for separation of small ions , DNA , peptides , proteins , and membrane vesicles as well as cells . Due to its complete liquid‐phase electrophoresis, FFE has numerous advantages, such as being with quite gentle and controllable experiment conditions, relatively high‐yield preparation, well protection of sample activity, and wholly continuous separation mode in contrast to a batch one in chromatography .…”

Reassemblable quasi‐chip free‐flow electrophoresis with simple heating dispersion for rapid micropreparation of trypsin in crude porcine pancreatin

“…These merits bequeath FFE with widespread potential application to bioseparation [13][14][15][16]. For example, Lanz et al [17] and Gratz et al [18] carried out the enantiomeric separation via LS-FFE. Loseva et al [19], Kasicka et al [20], and Malmstrom et al [21] performed a series of significant investigations on separation of proteins or peptides via LS-FFE.…”

Chiral Separations

“…Continuous-free flow electrophoresis was used for preparative chiral separations of piperoxan with sulfated -CD by Stalcup et al (186), and this method was again used with dyes to investigate the migration of the chiral selector and its impact on the chiral separation (187). Chiral separations utilizing ultrafiltration with a stereospecific binding agent in free solution were performed using bovine serum albumin as the binding agent for the analyte tryptophan (188).…”