A reversed phase HPLC assay for the simultaneous quantitation of non-ionic and ionic surfactants in bioprocess intermediates.
J Pharm Biomed Anal
; 40(4): 964-70, 2006 Mar 03.
Article
em En
| MEDLINE
| ID: mdl-16242286
ABSTRACT
This report describes a rapid and accurate reversed phase HPLC method for the simultaneous quantitation of multiple surfactants in various bioprocess solution matrices including cell lysates. Separation and quantitation of a mixture of the cationic detergent domiphen bromide from the non-ionic detergent Triton X-100 in crude cell mixtures can be achieved within 15 min using a TSK-gel C18-NPR reversed phase column and an aqueous mobile phase gradient of acetonitrilewater with the reagent PIC-B8 as ion-pairing modifier. The linear dynamic range for quantitation of domiphen bromide (DB) and Triton in this assay extends from 20 to 2000 microM. Linear regression analyses from the standard curve determinations showed an R2 of > or = 0.990. The assay does not show any interferences from proteins or other cellular contaminants such as nucleic acids. The assay has been used to evaluate clearance of these compounds throughout the purification process of an adenovirus-based vaccine candidate, as well as to determine the effects of process changes on detergent clearance.
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Base de dados:
MEDLINE
Assunto principal:
Tensoativos
/
Cromatografia Líquida de Alta Pressão
/
Vacinas de DNA
Idioma:
En
Revista:
J Pharm Biomed Anal
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
Estados Unidos