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the migration time of compounds that have a charge opposite of the micelle. Other alkyl surfactants that have gained popularity are the alkyl glycosides. Micelles composed of alkyl glycosides have been used in conjunction with borate buffers, and under these alkaline conditions in situ charged micelles are formed. Varying the ionic strength and the pH of the borate in the run buffer can change the density of these micelles. El Rassi and coworkers have been successful with these types of separations.62 64 A list of commonly used alkyl surfactants and the CMC of each is given in Table 8.1. Bile salts are anionic surfactants that can be found in biological sources. They have steroidal structures and can form reverse micelles.65 Bile salt surfactants are typically used to separate extremely hydrophobic.34 40 Bile salts are an excellent alternative choice when compounds do not separate and migrate with the tmc of a SDS run buffer system. Figure 8.2 shows how changing the micelle from SDS to sodium cholate can affect the resolution of corticosteroids. The effect of the hydrophobicity of different bile salt can also be seen in Figure 8.3. Resolution of the same corticosteroids can be improved when replacing sodium cholate with sodium deoxycholate. Because of their unique structure various bile salts have demonstrated the ability to separate chiral compounds.32,35,66 68 Bile salt micelles can handle organic solvents better than the alkyl chain surfactants broadening the scope of their applications.65 The source of the bile salts is important to ensure a clean electropherogram, since they can contain impurities that interfere with UV detection at low wavelengths, potentially decreasing the sensitivity of the assay. Therefore several sources of bile salts should be considered to ensure that the highest purity salts are selected for the run buffer. Table 8.2 shows a list of the most commonly used bile salts. Adding an organic modi er to the run buffer41,69 71 can enhance the separation of neutral and hydrophobic molecules. Organic solvents have been used widely to affect the selectivity of HLPC separations. They usually obtain optimal resolution, reasonable retention times, and help remove compounds that are very hydrophobic and highly retained by the stationary phase. Increasing the concentration of the organic modi er causes the compounds to elute at faster retention times. However, when an organic modi er is added to the run buffer, a reverse effect is observed with MEKC. When the organic modi er is added to the run buffer, it increases the solubility of the compounds in the aqueous portion of the run buffer, causing them to migrate at a time faster than the tmc. Besides changing the resolution adding an organic modi er decreases the electroosmotic ow of the system. This creates a larger migration window, which increases the analysis time of the system. The observed migration time appears to be directly proportional to the viscosity changes that come with using different organic
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Table 8.1. Long-Chain Alkyl Surfactants for MEKC
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Surfactant Cationic Decyltrimethylammonium bromide (DeTAB) Decyltrimethylammonium chloride (DeTAc) Dodecyltrimethylammonium bromide (DTAB) Dodecyltrimethylammonium chloride (DTAC) Tetradecylammonium bromide (TTAB) Tetradecylammonium chloride (TTAC) Cetyltrimethylammonium bromide (CTAB) Cetyltrimethylammonium chloride (CTAC) Hexadecyltrimethylammonium bromide (HTAB) Hexadecyltrimethylammonium chloride (HTAC) Anionic Sodium dodecyl sulfate (SDS) sodium tetradecyl sulfate (STS) Sodium decanesulfonate Sodium dodecanesulfonate Sodium N-lauroyl-N-methyltaurate (LMT) Sodium polyoxythylene(3) dedecyl ether sulfate Potassium per uoraoheptanoate Sodium tetradecene sulfonate (OS-14) Sodium polyoxyethylene(3) dodecyl ether acetate Sodium di-2-ethylhecyl sulfosuccinate Disodium 5,12-bis(doedecyloxymethyl)-4,7,10,13 -tetraoxa-1,16-hexadecanedisulfonate Per uorooctanesulfonate 3b-glucopyranosyl-5b-cholan-12a-hydroxy-24-oic acid Neutral Polyoxyethylene(6) dodecyl ether Polyoxyethylene(23) dodecyl ether (brij 35) Polyoxyethylene (20) sorbitanmonolaurate (tween 20) Polyoxyethylene (20) sorbitanmonooctadecanate (tween 60) Heptyl-b-d-glucopyranoside Octyl-b-d-glucopyranoside Nonyl-b-d-glucopyranoside Decyl-b-d-glucopyranoside Octanoyl-N-methylglucamide Nonanoyl-N-methylglucamide Decanoyl-N-methylglucamide Octyl-b-maltopyranoside n-Octanoylsucrose
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