Analytical and Bioanalytical Chemistry

, Volume 389, Issue 6, pp 1869–1878 | Cite as

Rapid determination of pK a values of 20 amino acids by CZE with UV and capacitively coupled contactless conductivity detections

  • Yveline Henchoz
  • Julie Schappler
  • Laurent Geiser
  • Josiane Prat
  • Pierre-Alain Carrupt
  • Jean-Luc Veuthey
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

A rapid and universal capillary zone electrophoresis (CZE) method was developed to determine the dissociation constants (pK a) of the 20 standard proteogenic amino acids. Since some amino acids are poorly detected by UV, capacitively coupled contactless conductivity detection (C4D) was used as an additional detection mode. The C4D coupling proved to be very successful on a conventional CE-UV instrument, neither inducing supplementary analyses nor instrument modification. In order to reduce the analysis time for pK a determination, two strategies were applied: (i) a short-end injection to reduce the effective length, and (ii) a dynamic coating procedure to generate a large electroosmotic flow (EOF), even at pH values as low as 1.5. As a result, the analysis time per amino acid was less than 2 h, using 22 optimized buffers covering a pH range from 1.5 to 12.0 at a constant ionic strength of 50 mM. pK a values were calculated using an appropriate mathematical model describing the relationship between effective mobility and pH. The obtained pK a values were in accordance with the literature.

Figure

a UV (1) and C4D (2) detectors placed on-line on the CE capillary. b Curve of effective mobility as a function of pH for histidine

Keywords

Amino acids Capillary zone electrophoresis Capacitively coupled contactless conductivity detection Dissociation constants Dynamic coating 

Abbreviations

ADME

Absorption, distribution, metabolism and excretion

BGE

Background electrolyte

C4D

Capacitively coupled contactless conductivity detection

CE

Capillary electrophoresis

CZE

Capillary zone electrophoresis

EOF

Electroosmotic flow

FS

Fused silica

ID

Internal diameter

NCE

New chemical entity

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Yveline Henchoz
    • 1
    • 2
  • Julie Schappler
    • 1
  • Laurent Geiser
    • 1
  • Josiane Prat
    • 1
  • Pierre-Alain Carrupt
    • 2
  • Jean-Luc Veuthey
    • 1
  1. 1.Laboratoire de Chimie Analytique Pharmaceutique, Section des sciences pharmaceutiquesUniversité de Genève, Université de LausanneGeneva 4Switzerland
  2. 2.Unité de Pharmacochimie, Section des sciences pharmaceutiquesUniversité de Genève, Université de LausanneGeneva 4Switzerland

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