Development and validation of a rapid reversed-phase HPLC method for the determination of insulin from nanoparticulate systems

• Tipo: Artigo em Revista Científica Internacional
• Data: 2006

Autores

Bruno Sarmento (Autor) Outra; Antonio Ribeiro (Autor) Outra; Francisco Veiga (Autor) Outra; Domingos Ferreira (Autor) FFUP

Revista

Título: Biomedical Chromatography

Vol. 20

Páginas: 898-903

ISSN: 0269-3879

Editora: Wiley-Blackwell

Indexação

ISI Web of Knowledge - 62 Citações

Scopus - 79 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Química

Outras Informações

ID Authenticus: P-004-HCP

DOI: 10.1002/bmc.616

Abstract (EN): A reversed-phase high-performance liquid chromatographic (HPLC) method has been developed and validated for the determination of insulin in nanoparticulate dosage forms. Its application for the development and characterization of insulin-loaded nanoparticulates composed of polyelectrolytes has also been carried out. A reversed-phase (RP) C-18 column and gradient elution with a mobile phase composed of acetonitrile (ACN) and 0.1% aqueous trifluoroacetic acid (TFA) solution at a flow rate of 1 mL/min was used. Protein identification was made by UV detection at 214 nm. The gradient changed from 30:70 (ACN:TFA, v/v) to 40:60 (v/v) in 5 min followed by isocratic elution at 40:60 (v/v) for a further five minutes. The method was linear in the range of 1-100 mu g/mL (R-2 = 0.9996), specific with a good inter-day and intra-day precision based on relative standard deviation values (less than 3.80%). The recovery was between 98.86 and 100.88% and the detection and quantitation limits were 0.24 and 0.72 mu g/mL, respectively. The method was further tested for the determination of the association efficiency of insulin to nanoparticulate carriers composed of alginate and chitosan, as well as its loading capacity for this protein. Encapsulant release under simulated gastrointestinal fluids was evaluated. The method can be used for development and characterization of insulinloaded nanoparticles made from cross-linked chitosan-alginate. Copyright (c) 2006 John Wiley & Sons, Ltd.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 6