Capillary zone electrophoresis of pharmaceutical peptides

Langenhuizen, Marleen H.J.M.; Janssen, P. S. L.

doi:10.1016/0021-9673(93)83443-v

Cited by 21 publications

(5 citation statements)

References 13 publications

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“…In CE, a commonly solution to this is a use of a low pH [11] or high ionic strength buffer, sometimes with an organic modifier [12,13], and a covalent (static) [14] or dynamic coating [14][15][16][17] modification of the capillary surface. Recently, there have been many reviews on this subject [18,19].…”

Section: Introductionmentioning

confidence: 99%

Cationic amylopectin derivatives as additives for analysis of proteins in capillary electrophoresis

Kato

Sakai‐Kato

et al. 2006

Electrophoresis

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Positively charged amylopectin, which is a major constituent of cationic starch, was used to modify the inner surface of fused-silica capillaries by addition to the running solution, which was subsequently employed in CE. Capillaries filled with cationic amylopectin derivatives were shown to generate a stable reversed EOF in the investigated range of pH 4-8. Among the additives studied, quaternary ammonium amylopectin derivatives with high amino and low hydroxypropyl groups showed fast electroosmotic mobility and very effectively suppressed the adsorption of proteins. The run-to-run and batch-to-batch repeatability of the procedures were satisfactory with RSDs of 0.5% and 2.4%, respectively. A basic protein, alpha-chymotrypsinogen, migrated within 6 min and the theoretical plate number of it reached 560 000 plates/m.

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Section: Introductionmentioning

confidence: 99%

Cationic amylopectin derivatives as additives for analysis of proteins in capillary electrophoresis

Kato

Sakai‐Kato

et al. 2006

Electrophoresis

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“…These factors may reduce the resolution to an unacceptable level. The use of a low-pH buffer sometimes with an organic modifier is one of the useful methods for the separation of cationic analytes. − The acidic pH suppresses the dissociation of silanol groups on the inner surface of the capillary and allows peptide or protein separations with minimum wall interactions. Another challenge in implementing CE/MS is that the running buffer must bring about an adequate CE separation of peptides without disturbing the MS signal.…”

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confidence: 99%

Integration of On-Line Protein Digestion, Peptide Separation, and Protein Identification Using Pepsin-Coated Photopolymerized Sol−Gel Columns and Capillary Electrophoresis/Mass Spectrometry

et al. 2004

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A miniaturized pepsin reactor was prepared inside a fused-silica capillary (i.d. 75 microm) by coating a pepsin-containing gel on a photopolymerized porous silica monolith. The pepsin-encapsulated film was prepared by a sol-gel method. The sol-gel reaction was optimized so that the sol solution containing pepsin forms a thin film on the photopolymerized sol-gel (PSG) monolith that was initially fabricated at the inlet of the capillary. Pepsin was encapsulated into the gel matrix without losing its activity. The large surface area of the PSG monolith enabled the immobilized pepsin to achieve a high catalytic turnover rate, and the porous nature of the PSG promotes penetration of large molecular proteins into the column. The immobilized pepsin-digested peptides and proteins, and the resulting mixture of peptide fragments, could be directly separated in the portion of the capillary where no PSG monolith exists. The durability and repeatability of the fabricated pepsin-coated column was tested and found to be satisfactory. An acidic solution consisting of 0.5 M formic acid was used as the running buffer, because it suppresses the adsorption of proteins or peptides on the inner surface of the capillary as well as enables direct connection of the output of the capillary electrophoresis column to a mass spectrometer. The on-line digestion of insulin chain beta and lysozyme provides identification of the proteolytic peptides. Recovery was achieved for 100% of the insulin chain beta amino acid sequence and 73% of the lysozyme amino acid sequence.

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“…Peptide Separation and Detection Methodology. Protocols for manipulation of pH and buffer concentration for peptide separation methods were described (480). The separation of dipeptides and small peptides by various modes of capillary electrophoresis was investigated to identify the best separation conditions and to compare the different charge to size parameters used in correlating peptide migration.…”

Section: Applicationsmentioning

confidence: 99%