Full-capillary sample stacking/sweeping-MEKC for the separation of naphthalene-2,3-dicarboxaldehyde-derivatized tryptophan and isoleucine.
In an attempt to improve the sensitivity of detection in capillary electrophoresis (CE), a novel online sample-concentration method, full-capillary sample stacking (FCSS)/sweeping-micellar electrokinetic chromatography (sweeping-MEKC) mode, is proposed. Naphthalene-2,3-dicarboxaldehyde (NDA)-derivatized tryptophan and isoleucine were selected as model compounds. In the initial step, the weakly acidic compounds, dissolved in a low-conductivity buffer (35.1 microS/cm; apparent ph (pH*) in a mixed solution of acetonitrile/methanol/water, 4.6), fill the entire capillary, two vials of a high-conductivity buffer (2.06 mS/cm; pH* 2.0) are placed on each end, and a negative polarity is then applied. Under these conditions, the direction of the electroosmotic flow (EOF) is toward the inlet. Meanwhile, the anionic analytes move in the reverse direction and are neutralized and stacked at the boundary of a dynamic pH-junction (between the sample matrix and the nonmicellar background solution (BGS)). When the sample concentration is completed, the BGS is quickly changed to solutions containing SDS-BGS for the subsequent separation. Since the mobility of SDS-analytes is then greater than the EOF, the following steps occur by the sweeping (for focusing) and MEKC (for separation) mode. Using these steps, a full-capillary sample injection/separation can be achieved.
Shih
Chun-Min
CM
Department of Chemistry, National Taiwan Normal University, Taipei, Taiwan.
Lin
Cheng-Huang
CH
D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D007532:Isoleucine / Q000031:analogs & derivatives* / Q000302:isolation & purification*; D009281:Naphthalenes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000302:isolation & purification*
D009281:Naphthalenes; D007532:Isoleucine; C027717:2,3-naphthalenedicarboxaldehyde; D014364:Tryptophan