Assay of protein drug substances present in solution mixtures by fluorescamine derivatization and capillary electrophoresis.
A method is described to enhance the resolution and detection sensitivity of proteins, peptides, and amino acids in capillary electrophoretic analysis of solution mixtures. The method consists of derivatizing the analytes with fluorescamine, which is normally used as a fluorogenic reagent for compounds containing a reactive primary amine functional group, and then using the derivative as an ultraviolet chromophore to enhance detection sensitivity (measured at 280 nm) in capillary electrophoresis. The results demonstrated a significant improvement in the separation and detection sensitivity of the derivatized analytes as compared to their underivatized counterparts. The use of chromophores, such as fluorescamine, in capillary electrophoresis facilitates the analysis of components of solution mixtures, such as pharmaceutical formulations, that could not be resolved and/or detected by conventional capillary electrophoresis procedures.
Guzman
N A
NA
Pharmaceutical Research and Development, Hoffmann-La Roche, Nutley, NJ 07110.
Moschera
J
J
Bailey
C A
CA
Iqbal
K
K
Malick
A W
AW
D000911:Antibodies, Monoclonal; D004586:Electrophoresis / Q000379:methods; D005450:Fluorescamine / Q000737:chemistry; D007370:Interferon Type I / Q000032:analysis; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D011994:Recombinant Proteins; D013056:Spectrophotometry, Ultraviolet
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D004364:Pharmaceutical Preparations; D011506:Proteins; D011994:Recombinant Proteins; D005450:Fluorescamine