title: A versatile, stability-indicating and high-throughput ultra-fast liquid chromatography method for the determination of isoflavone aglycones in soybeans, topical formulations, and permeation assays.
abstract: There is a growing interest in the pharmaceutical field concerning isoflavones topical delivery systems, especially with regard to their skin care properties and antiherpetic activity. In this context, the present work describes an ultra-fast liquid chromatography method (UFLC) for determining daidzein, glycitein, and genistein in different matrices during the development of topical systems containing isoflavone aglycones (IA) obtained from soybeans. The method showed to be specific, precise, accurate, and linear (0.1 to 5 µg mL(-1)) for IA determination in soybean acid extract, IA-rich fraction obtained after the purification process, IA loaded-nanoemulsions, and topical hydrogel, as well as for permeation/retention assays in porcine skin and porcine esophageal mucosa. The matrix effect was determined for all complex matrices, demonstrating low effect during the analysis. The stability indicating UFLC method was verified by submitting IA to acidic, alkaline, oxidative, and thermal stress conditions, and no interference of degradation products was detected during analysis. Mass spectrometry was performed to show the main compounds produced after acid hydrolysis of soybeans, as well as suggest the main degradation products formed after stress conditions. Besides the IA, hydroxymethylfurfural and ethoxymethylfurfural were produced and identified after acid hydrolysis of the soybean extract and well separated by the UFLC method. The method's robustness was confirmed using the Plackett-Burman experimental design. Therefore, the new method affords fast IA analysis during routine processes, extract purification, products development, and bioanalytical assays.
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Marina C
authors.initials: MC
authors.lastname: Nemitz
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Francini K J
authors.initials: FKJ
authors.lastname: Yatsu
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Juliana
authors.initials: J
authors.lastname: Bidone
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Letícia S
authors.initials: LS
authors.lastname: Koester
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Valquiria L
authors.initials: VL
authors.lastname: Bassani
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Cássia V
authors.initials: CV
authors.lastname: Garcia
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Pampa (UNIPAMPA), BR 472 Km 585 Prédio Administrativo CEP 97500-970, Uruguaiana, RS, Brazil.
authors.forename: Andreas S L
authors.initials: ASL
authors.lastname: Mendez
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil.
authors.forename: Gilsane L
authors.initials: GL
authors.lastname: von Poser
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga 2752, CEP 90610-000, Porto Alegre, RS, Brazil. Electronic address: helder.teixeira@ufrgs.br.
authors.forename: Helder F
authors.initials: HF
authors.lastname: Teixeira
mesh_terms: D000287:Administration, Topical; D000818:Animals; D001692:Biological Transport; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004947:Esophagus / Q000187:drug effects / Q000378:metabolism; D005662:Furaldehyde / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D019833:Genistein / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D020100:Hydrogels; D006868:Hydrolysis; D007529:Isoflavones / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D009092:Mucous Membrane / Q000187:drug effects / Q000378:metabolism; D010539:Permeability; D010936:Plant Extracts / Q000737:chemistry; D012867:Skin / Q000187:drug effects / Q000378:metabolism; D013025:Glycine max / Q000737:chemistry*; D013552:Swine
chemical_list: D020100:Hydrogels; D007529:Isoflavones; D010936:Plant Extracts; C004742:daidzein; C008046:5-hydroxymethylfurfural; C086566:glycitein; D019833:Genistein; D005662:Furaldehyde
keywords: Isoflavone aglycones; Matrix effect; Permeation/retention assays; Soybean acid extract; Stability indicating UFLC method; Topical delivery systems