title: Development and validation of a rapid HPLC method for the quantification of GSE4 peptide in biodegradable PEI-PLGA nanoparticles.
abstract: In this work a high performance liquid chromatographic (HPLC) method has been developed and validated for the content determination of GSE4 peptide in PEI-PLGA nanoparticles. Chromatographic separation was performed on a C18 column, and a gradient elution with a mobile phase composed of methanol and 0.1% aqueous trifluoroacetic acid (TFA) solution, at a flow rate of 1ml/min, was used. GSE4 peptide identification was made by fluorescence detection at 290nm. The elution of methanol:TFA was initially maintained at (20:80, v/v) for one min and the gradient changed to (80:20, v/v) in 6min. This ratio was then followed by isocratic elution at (80:20, v/v) during another min and for further 3min it was linearly modified to (20:80, v/v). The developed method was validated according to the ICH guidelines, being specific, linear in the range 10-100μg/ml (R(2)=0.9996), precise, exhibiting good inter-day and intra-day precision reflected by the relative standard deviation values (less than 3.88%), accurate, with a recovery rate of 100.18±0.95%, and stable for 48h at 5°C or at RT when encapsulated in nanoparticles. The method was simple, fast, and successfully used to determine the peptide content in GSE4-loaded PEI-PLGA nanoparticles.
authors.affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country, Faculty of Pharmacy, Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain. Electronic address: susanapatricia.egusquiaguirre@ehu.es.
authors.forename: Susana P
authors.initials: SP
authors.lastname: Egusquiaguirre
authors.affiliation: Instituto de Investigaciones Biomédicas CSIC/UAM, IDIPaz and CIBER de Enfermedades Raras CIBERER, Arturo Duperier, 4, 28029 Madrid, Spain. Electronic address: cmanguan@iib.uam.es.
authors.forename: Cristina
authors.initials: C
authors.lastname: Manguán-García
authors.affiliation: Instituto de Investigaciones Biomédicas CSIC/UAM, IDIPaz and CIBER de Enfermedades Raras CIBERER, Arturo Duperier, 4, 28029 Madrid, Spain. Electronic address: rperona@iib.uam.es.
authors.forename: Rosario
authors.initials: R
authors.lastname: Perona
authors.affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country, Faculty of Pharmacy, Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain. Electronic address: joseluis.pedraz@ehu.es.
authors.forename: José Luís
authors.initials: JL
authors.lastname: Pedraz
authors.affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country, Faculty of Pharmacy, Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain. Electronic address: rosa.hernandez@ehu.es.
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Hernández
authors.affiliation: NanoBioCel Group, Laboratory of Pharmaceutics, University of the Basque Country, Faculty of Pharmacy, Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain; Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Paseo de la Universidad, 7, 01006 Vitoria-Gasteiz, Spain. Electronic address: manoli.igartua@ehu.es.
authors.forename: Manuela
authors.initials: M
authors.lastname: Igartua
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019344:Lactic Acid / Q000737:chemistry*; D053758:Nanoparticles; D010455:Peptides / Q000032:analysis*; D011094:Polyethyleneimine / Q000737:chemistry*; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D015203:Reproducibility of Results
chemical_list: D010455:Peptides; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D011094:Polyethyleneimine
keywords: GSE4; HPLC; Nanoparticles; PEI; PLGA; Validation