Spermine-graft-dextran non-covalent copolymer as coating material in separation of basic proteins and neurotransmitters by capillary electrophoresis

J Chromatogr A. 2010 Jul 30;1217(31):5130-6. doi: 10.1016/j.chroma.2010.06.019. Epub 2010 Jun 11.

Abstract

Spermine-graft-dextran (Spe-g-Dex) copolymer was synthesized and used as a non-covalent coating for the separation of proteins and neurotransmitters by capillary electrophoresis. The coating was obtained via flushing the capillary with 1.0% Spe-g-Dex copolymer solution for 2min. Electroosmotic flow (EOF) was strongly suppressed, ranging from -1.60x10(-9) to 3.65x10(-9)m(2)V(-1)s(-1). Effect of experimental conditions, such as the copolymer concentration, the concentration and pH of the background electrolyte (BGE), on the Spe-g-Dex coating was investigated. Separation of lysozyme, cytochrome c, ribonuclease A and alpha-chymotrypsinogen yielded high separation efficiencies ranging from 141000 to 303000plates/m and recoveries from 85.4% to 98.3% at pH 4.0 (284.0mM sodium acetate-acetic acid buffer, I=50mM). Run-to-run repeatabilities and day-to-day, and capillary-to-capillary reproducibilities were all below 1.7%. In addition, Spe-g-Dex coating allowed the successful separation of five neurotransmitters, 5-hydroxytryptamine, dopamine, epinephrine, isoprenaline, dobuamine at pH 4.0 with high separation efficiencies of 290000-449000plates/m.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dextrans / chemistry
  • Electrophoresis, Capillary / instrumentation*
  • Electrophoresis, Capillary / methods*
  • Neurotransmitter Agents / analysis
  • Neurotransmitter Agents / isolation & purification*
  • Polymers / chemistry*
  • Protein Binding
  • Proteins / analysis
  • Proteins / isolation & purification*
  • Spermine / chemistry

Substances

  • Dextrans
  • Neurotransmitter Agents
  • Polymers
  • Proteins
  • Spermine