Amino acids electrolytic separation

Reverse miceUes have been applied in the separation of amino acids and proteins. The separation is based on the balance between electrostatic forces and hydrophobic interactions [120]. The pH value is a crucial parameter determining this balance. If reversed miceUes are applied in LMs, then the underlying interactions are determined by interfacial partition coefficients of the amino acids/proteins separated, that is, hydrophobicity of the compounds separated, ionic strength of the feed and stripping solutions, the chemical nature of the electrolytes present, and the intertacial curvature of the amphiphilic film [121]. Changing the above-mentioned conditions, the overaU charge of the reverse miceUe can be altered, and so the separation conditions can be manipulated [122]. 

Patients who can only be fed parenterally over a longer pcricxl of lime need not only calories but also amino acids, electrolytes, trace elements, and vitamins. However, if separate infusions of the different solutions are applied very large volume.s (up to 5 L) must be given. For this reason so-called all-in-one solution.s are aimed at, A popular standard formulation (=2500 kcal. 2600 mL) consists of HKHl-mL ammo acid solution (isoleucinc. leucine, lysine, methionine, phenylalanine. threonine, tryptophane, valine, alanine, arginine, histidine, prolinc. tyro-... 

Table 3-1. Values of enantiomeric resolution of DNP-amino acids in a running electrolyte containing the three fractions 1, 2, and 3 of the cyclo(Arg-Lys-X-Pro-X-(3 Ala) sublibrary separated by preparative HPLC.

The gradient in IEF is a pH gradient that is generated by electrolyte systems consisting of ampholytes. The capillary wall is coated in order to avoid electroosmosis. The IEF mode is especially suitable for the separation of zwitter ionic compounds like proteins, peptides, amino acids, various drugs, etc. After injection of the sample and application of the potential, the sample constituents migrate to a pH in the capillary... 

CE has been applied extensively for the separation of chiral compounds in chemical and pharmaceutical analysis.First chiral separations were reported by Gozel et al. who separated the enantiomers of some dansylated amino acids by using diastereomeric complex formation with Cu " -aspartame. Later, Tran et al. demonstrated that such a separation was also possible by derivatization of amino acids with L-Marfey s reagent. Nishi et al. were able to separate some chiral pharmaceutical compounds by using bile salts as chiral selectors and as micellar surfactants. However, it was not until Fanali first showed the utilization of cyclodextrins as chiral selectors that a boom in the number of applications was noted. Cyclodextrins are added to the buffer electrolyte and a chiral recognition may... 

Wang and Porter [92] resolved the enantiomers of oxazepam, lorazepam, and temazepam using /1-cyclodextrin as the CMPA by CEC. The authors varied separation parameters such as voltage and mobile phase. Wei et al. [93] resolved the enantiomers of phenylephrine and synephrine by varying the concentration of /1-cyclodextrin (CMPA), pH, electrolyte concentration, and temperature. Lelievre et al. [99] separated the enantiomers of chlorthalidone using hydroxypropyl /1-cyclodextrin as the CMPA. Lammerhofer and Lindner [90] resolved the enantiomers of N-derivatized amino acids (e.g., 3,5-dinitrobenzoyl, 3,5-dinitro-benzyloxycarbonyl, 2,4-dinitrophenyl, and 9-fluorenylmethoxycarbonyl amino... 

J.N. Weinstein and S.R. Caplan, Charge-mosaic Membranes Dialydc Separation of Electrolytes from Nonelectrolytes and Amino Acids, Science 296, 169 (1970). 

Lammerhofer and Lindner [62] reported on the enantiomer separation of deriva-tized amino acids on an ODS-packed capillary with a chiral quinine-derived selector as buffer additive in two different modes (i) in an electrophoretically dominated mode at high electrolyte concentration and (ii) in an electroosmotically dominated mode at a low electrolyte concentration. Enantiomer separation in the electrophoretically dominated mode (i) leads to high efficieny (about two to three times higher than in LC) but to a moderate enantioselectivity (about the same as in LC). In the electroosmotically dominated mode (ii) a higher enantioselectivity but a lower efficiency (even inferior to LC) occurs. The separations can also been performed in a non-aque-ous buffered mobile phase. Pressurization (8-10 bar) of the flow system on both ends of the separation capillary was applied. 

Capillary zone electrophoresis (CZE), micellar capillary electrokinetic chromatography (MECC), capillary gel electrophoresis (CGE), and affinity capillary electrophoresis (ACE) are CE modes using continuous electrolyte solution systems. In CZE, the velocity of migration is proportional to the electrophoretic mobilities of the analytes, which depends on their effective charge-to-hydrodynamic radius ratios. CZE appears to be the simplest and, probably, the most commonly employed mode of CE for the separation of amino acids, peptides, and proteins. Nevertheless, the molecular complexity of peptides and proteins and the multifunctional character of amino acids require particular attention in selecting the capillary tube and the composition of the electrolyte solution employed for the separations of these analytes by CZE. 

Enantiomeric separations of amino acids and short peptides are performed using either a direct or the indirect approach [10]. The indirect approach employs chiral reagents for diasteromer formation and their subsequent separation by various modes of CE. The direct approach uses a variety of chiral selectors that are incorporated into the electrolyte solution. Chiral selectors are optically pure compounds bearing at least one functional group with a chiral center (usually represented by an asymmetric carbon atom) which allows sterically selective interactions with the two enantiomers. Among others, cyclodextrins (CDs) are the... 

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