Separation and quantitative analysis of alkyl sulfobetaine-type detergents by high-performance liquid chromatography and light-scattering detection - Analytical Methods & Tools for Lyophilization

Separation and quantitative analysis of alkyl sulfobetaine-type detergents by high-performance liquid chromatography and light-scattering detection
Received 10 March 2006; accepted 22 August 2006. Available online 15 September 2006.
Laurence Zannaa and Jean-Fran?ois Haeuw
Journal of Chromatography B
Article in Press
ScienceDirect
Copyright ? 2006 Published by Elsevier B.V.
aCentre d?Immunologie Pierre Fabre, 5 Avenue Napol?on III, BP 497, 74160 Saint-Julien-en-Genevois, France
Abstract
An improved high-performance liquid chromatographic (HPLC) method for the separation of zwitterionic detergents is described. It is based on a reversed-phase liquid chromatography with evaporative light-scattering detection (ELSD). The method was shown to be highly specific, allowing the separation of three detergents of the alkyl sulfobetaine family: 3-(N-dodecyl-N,N-dimethyl-ammonio)-propane-1-sulfonate (SB12), 3-(N-tetradecyl-N,N-dimethyl-ammonio)-propane-1-sulfonate (SB14) and 3-(N-hexadecyl-N,N-dimethyl-ammonio)-propane-1-sulfonate (SB16). It was further used to develop a quantitation method for SB14, which was validated for linearity, precision, robustness, limits of detection and quantitation, specificity and accuracy. Linearity was found in the range of 50?500 ?g/ml with a correlation coefficient of 0.9938 ? 0.0029. The mean value of slope and intercept were 1.567 ? 0.06 and 0.1541 ? 0.0271, respectively. The limits of detection (LOD) and quantitation (LOQ) were 2 and 10 ?g/ml, respectively. The validated method was used to determine the concentration of SB14 in different biological samples, specially in bulks of a recombinant membrane protein, the Klebsiella pneumoniae outer membrane protein A, which is produced at the pilot scale for human clinical studies.
Keywords: High-performance liquid chromatographic (HPLC); Evaporative light-scattering detection (ELSD); Detergent; Zwitterionic; Alkyl sulfobetaine; Surfactant
Corresponding author. Tel.: +33 4 50 35 35 54; fax: +33 4 50 35 35 90.
You can view the abstract online. A subscription is required to view the full text or it can be purchased online.