Comparative study on sample stacking by moving reaction boundary formed with weak acid and weak or strong alkali in...

The condensation of low abundance zwitterion substance, such as protein and peptide, has great significance to the study on proteomics. This paper develops the theory on design of online stacking conditions of zwitterion by a moving reaction boundary (MRB) in capillary electrophoresis (CE). This concerns the choice of running and sample buffers, velocity design of MRB, and salt effect on the stacking. The theoretical results unveil that: (1) the velocity of MRB formed with weak acidic buffer and strong alkali should be set between zero and the velocity of zwitterion in the alkali phase, or no stacking occurs; (2) if a strong alkali is used to prepare the sample, a much long front plug of strong base must be injected before the alkaline sample plug for complete stacking, whereas no such front plug is needed if a weak alkali with enough high concentration and pH value is used to prepare the sample buffer; (3) the existence of salt in sample matrix has a weak effect on the stacking of zwitterion if sample is prepared with weak alkaline buffer, while has a dramatic effect on the same stacking if with a strong base buffer. In addition, the concentration of weak alkali used for preparation of sample should be set at the point, at which the velocity of MRB is as much as possible close to that of negative zwitterion. The developed theory and its computation are quantitatively proved by the experiments of zwitterion stacking by the MRB as shown in the previous and the accompanying papers. The proposed theoretic results hold obvious significances on-column stacking of low abundance zwitterions, such as amino acid, or peptides or proteins, in CE.