Nemitz Marina C, Yatsu Francini K J, Bidone Juliana, Koester Letícia S, Bassani Valquiria L, Garcia Cássia V, Mendez Andreas S L, von Poser Gilsane L, Teixeira Helder F
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.
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Pampa (UNIPAMPA), BR 472 Km 585 Prédio Administrativo CEP 97500-970, Uruguaiana, RS, Brazil.
Talanta. 2015 Mar;134:183-193. doi: 10.1016/j.talanta.2014.10.062. Epub 2014 Nov 11.
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.
在制药领域，人们对异黄酮局部给药系统的兴趣与日俱增，尤其是在其护肤特性和抗疱疹活性方面。在此背景下，本研究描述了一种超快速液相色谱法（UFLC），用于在开发含大豆异黄酮苷元（IA）的局部给药系统过程中，测定不同基质中的大豆苷元、染料木黄酮和黄豆黄素。该方法在测定大豆酸提取物、纯化后得到的富含IA的馏分、负载IA的纳米乳液和局部水凝胶中的IA时，以及在猪皮肤和猪食管黏膜的渗透/滞留试验中，显示出特异性、精密度、准确性和线性（0.1至5μg mL⁻¹）。测定了所有复杂基质的基质效应，结果表明分析过程中基质效应较低。通过将IA置于酸性、碱性、氧化和热应激条件下，验证了该稳定性指示UFLC方法，分析过程中未检测到降解产物的干扰。进行了质谱分析，以显示大豆酸水解后产生的主要化合物，并推测应激条件下形成的主要降解产物。除了IA外，大豆提取物酸水解后还产生并鉴定出羟甲基糠醛和乙氧基甲基糠醛，且它们能被UFLC方法很好地分离。使用Plackett-Burman实验设计证实了该方法的稳健性。因此，该新方法可在常规过程、提取物纯化、产品开发和生物分析测定中快速分析IA。
