Qualitative and quantitative reversed-phase high performance liquid chromatographic analysis of glycoprotein hormones in the presence of a large excess of human serum albumin

Autor(es)

Almeida, B. E. ^[1] ; Oliveira, J. E. ^[1] ; Damiani, R. ^[1] ; Dalmora, S. L. ^[2] ; Bartolini, P. ^[1] ; Ribela, M. T. C. P. ^[1]

Afiliação do(s) autor(es)

^[1] IPEN CNEN, Dept Biotechnol, BR-05508900 Sao Paulo - Brazil

^[2] Univ Fed Santa Maria, Dept Ind Pharm, BR-97119900 Santa Maria, RS - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Pharmaceutical and Biomedical Analysis; v. 63, p. 160-164, APR 7 2012.
• Citações Web of Science: 2

Resumo

The present work describes reversed-phase high performance liquid chromatographic methodologies (RP-HPLC) for the qualitative and quantitative analysis of the human glycoprotein hormones thyrotropin (hTSH), follitropin (hFSH), choriogonadotropin (hCG) and lutropin (hLH) in the presence of a large excess (up to 250:1) of human serum albumin (HSA). Chromatographic profiles with a good separation between the hormone and HSA were obtained by using a C4 column and specific gradient elution conditions for each hormone. Parameters such as resolution factor, tailing factor and relative retention time, were determined, and are useful for the evaluation of the quality of the separation obtained between the active pharmaceutical ingredient and the excipient present in the final formulation. The potential of each method for quantification of both HSA and the hormone was also demonstrated. Besides furnishing chromatographic quantifications that can substitute for in vivo bioassays and animal use, the chromatograms also provide a direct panorama of the quality and heterogeneity of the protein of interest. (C) 2012 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 11/07289-0 - Síntese e caracterização de diferentes glicoformas de tireotrofina humana recombinante (rhTSH) farmacologicamente ativas
• Beneficiário: Maria Teresa de Carvalho Pinto Ribela
• Modalidade de apoio: Auxílio à Pesquisa - Regular