Chromatography Research Today is a free monthly online journal that collates and summarizes the latest research about Chromatography, including details on column chromatography, gas chromatography (gc), liquid chromatograpy, hplc.

Quantitative aspects of the analysis of the monoclonal antibody trastuzumab using high-performance liquid chromatography coupled with electrospray mass spectrometry.

Damen CW, Rosing H, Schellens JH, Beijnen JH

Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066 EC Amsterdam, The Netherlands. carola.damen@slz.nl

The analytical possibilities of quantification of the intact monoclonal antibody trastuzumab by high-performance liquid chromatography coupled with electrospray mass spectrometry (HPLC-ESI-MS) were investigated. To clarify the results obtained by LC-MS, complementary experiments were performed using direct UV-spectrophotometry and high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV). A polystyrene-divinylbenzene (POROS) column was applied with gradient elution using formic acid 0.08% (v/v) in water and formic acid 0.08% (v/v) in acetonitrile as mobile phase for chromatographic analysis. Quantification on LC-MS was performed by using the peak area of the total ion current (TIC) chromatograms of one charge state. Non-linearity and sensitivity loss were the major limitations observed with the LC-MS method, of which the non-linearity is most likely caused by detector saturation. The sensitivity loss during analysis could be reduced by lowering the MS source temperature. This parameter is critical in creating a robust LC-MS system for the quantitative analysis of trastuzumab.

Published 14 January 2008 in J Pharm Biomed Anal, 46(3): 449-55.
Full-text of this article is available online (may require subscription).

© 2005-2008 Chromatography Research Today. All Rights Reserved.