Capillary electrophoresis buffer additives

Capillary Zone Electrophoresis The simplest form of capillary electrophoresis is capillary zone electrophoresis (CZE). In CZE the capillary tube is filled with a buffer solution and, after loading the sample, the ends of the capillary tube are placed in reservoirs containing additional buffer solution. Under normal conditions, the end of the capillary containing the sample is the anode, and solutes migrate toward... 

Tran, A. D., Park, S., Lisi, P. J., Huynh, O. T., Ryall, R. R., and Lane, P. A., Separation of carbohydrate-mediated microheterogeneity of recombinant human erythropoietin by free solution capillary electrophoresis. Effects of pH, buffer type and organic additives,. Ckromatogr., 542, 459, 1991. 

Lin et al. [95] used capillary electrophoresis with dual cyclodextrin systems for the enantiomer separation of miconazole. A cyclodextrin-modified micellar capillary electrophoretic method was developed using mixture of /i-cyclodextrins and mono-3-0-phenylcarbamoyl-/j-cyclodextrin as chiral additives for the chiral separation of miconazole with the dual cyclodextrins systems. The enantiomers were resolved using a running buffer of 50 mmol/L borate pH 9.5 containing 15 mmol/L jS-cyclodextrin and 15 mmol/L mono-3-<9-phcnylcarbamoyl-/j-cyclodextrin containing 50 mmol/L sodium dodecyl sulfate and 1 mol/L urea. A study of the respective influence of the /i-cyclodcxtrin and the mono-3-(9-phenylcarbamoyl-/i-cyclodextrin concentration was performed to determine the optical conditions with respect to the resolution. Good repeatability of the method was obtained. 

A high performance capillary electrophoresis (HPCE) was described for the separation and simultaneous determination of OTC, TC, CTC, DC, and chloramphenicol in honey. The use of buffer pH 3.2 containing 0.02 mol/L Na2HP04 and 0.01 mol/L citric acid with addition of 4% (v/v) A-methylmorpholine and 12% (v/v) acetonitrile demonstrated a good separation of these five antibiotics within 20 min. The proposed method gave detection limit (signal to noise ratio > 5) of 20 pg/L for OTC [26],... 

Anions and uncharged analytes tend to spend more time in the buffered solution and as a result their movement relates to this. While these are useful generalizations, various factors contribute to the migration order of the analytes. These include the anionic or cationic nature of the surfactant, the influence of electroendosmosis, the properties of the buffer, the contributions of electrostatic versus hydrophobic interactions and the electrophoretic mobility of the native analyte. In addition, organic modifiers, e.g. methanol, acetonitrile and tetrahydrofuran are used to enhance separations and these increase the affinity of the more hydrophobic analytes for the liquid rather than the micellar phase. The effect of chirality of the analyte on its interaction with the micelles is utilized to separate enantiomers that either are already present in a sample or have been chemically produced. Such pre-capillary derivatization has been used to produce chiral amino acids for capillary electrophoresis. An alternative approach to chiral separations is the incorporation of additives such as cyclodextrins in the buffer solution. 

Capillary electrophoresis (CE) instrument is quite simple. A CE, at its core, is merely a high-voltage power supply (capable of voltages in excess of 30,000 V), capillary (approximately 25 to 100 pm inner diameter), buffers to complete the circuit (e.g., citrate, phosphate, acetate, etc.), and a detector (e.g., UV-Vis). There are additional complexities, of course, but at its heart, the CE is a simple instrument. 

P Sun, N Wu, GE Barker, RA Hartwick. Chiral separations using dextran and bovine serum albumin as run buffer additives in affinity capillary electrophoresis. J Chromatogr 648 475-480, 1993. 

J Yang, DS Hage. Chiral separations in capillary electrophoresis using human serum albumin as a buffer additive. Anal Chem 66 2719-2725, 1994. 

In its simplest form capillary electrophoresis is termed capillary zone electrophoresis . The conditions used in this type of analysis are relatively simple and the mobile phase used consists of a buffer with various additives. Many applications focus on critical separations which are difficult to achieve by HPLC. In many cases it is difficult to explain completely the types of effects produced by buffer additives. 

PQQ was successfully separated from three closely related isomers by capillary electrophoresis with UV detection <2000JCH(876)193>. Rapid and efficient separation of all four compounds 17-20 as their negatively charged carboxylate ions with baseline resolution was achieved by the addition of 1-5 mM R4N salts to the capillary buffer. Detection limits of PQQ and its three isomers were in the range of 7-15 xM with mass detection limits of 98-210 fmol. 

Chiral crown ethers can be generally utilized as chiral selectors. They have been used as additives to mobile phases or running buffer in MECK and capillary electrophoresis (CE) systems124,125 (see Section 3.1.6.4.). 

The analysis of ILs may afford considerable insight into the physicochemical properties underlying the rich potential interaction chemistries of ILs [14] and suggest possibilities for future applications. Simultaneously, the unique features of ILs provide some intriguing new possibilities in the area of separations that have yet to be realized. Hence, topics to be covered in this chapter include analysis of ILs by LC, applications of ILs in liquid-phase microextraction (LPME), in high-performance LC (HPLC) as mobile-phase additives, and in capillary electrophoresis (CE) as buffer additives as well as applications of surface-confined ILs (SCIL) as novel stationary phases for LC. 

Marszalt, M. R, Markuszewski, M. J., and Kaliszan, R., Separation of nicotinic acid and its structural isomers using l-ethyl-3-methylimidazolium ionic liquid as a buffer additive by capillary electrophoresis, ]. Pharm. Biomed. Anal., 41, 329-332,2006. 

Alternatively, chiral separation can be performed by capillary electrophoresis (CE) with, for example, quaternary ammonium-/ -cyclodextrin as chiral buffer additive [184, 185]. 

Capillary electrophoresis (CE) provided an orthogonal separation technique. The retention mechanism can be manipulated with buffers or addition of surfactants to form micelles for the analysis where size and charge differences... 

One of the first steps in modifying the performance of capillary electrophoresis was the deactivation of silica groups of the capillary column by physically coating the capillary wall with methylcellulose (58,59), as well as via silane derivatization (10,44,60). Presently, many other changes have been carried out either to the capillary surface or addition of chemical agents to the separation buffer (see Table II), including manipulation of... 

TABLE II. Improvements on the Capillary Electrophoresis Separation of Proteins, Peptides and Amino Acids by Deactivation of the Silica Surface or by Addition of Chemical Agents to the Separation Buffer... 

Capillary electrophoresis (CE) provides a valid alternative to HPLC methods for chiral separations. The direct resolution of racemates requires only an enantiomerically pure additive (chiral selector) to be dissolved in the running buffer. The experimental conditions affecting the separations and an overview of practical applications have been compiled <1999ELP2605>. 

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