RESUMO
Fluorescent probes are used in drug nanocarrier pre-clinical studies or as active compounds in theranostics and photodynamic therapy. In the biological medium, nanoparticles interact with proteins, which can result in the off-target release of their cargo. The present study used asymmetric flow field-flow fractionation with online multi-angle laser light scattering and fluorescence detection (AF4-MALLS-FLD) to study the release, transfer, and partition of fluorescent dyes from polymeric nanoparticles (NP). NP formulations containing the dyes Rose Bengal, Rhodamine B, DiI, 3-(α-azidoacetyl)coumarin and its polymer conjugate, Nile Red, and IR780 and its polymer conjugate were prepared. NP suspensions were incubated in a medium with serum proteins and then analyzed by AF4. AF4 allowed efficient separation of proteins (< 10 nm) from fluorescently labeled NP (range of 54 - 180 nm in diameters). The AF4 analyses showed that some dyes, such as Rose Bengal, IR780, and Coumarin were transferred to a high extent (68-77%) from NP to proteins. By contrast, for DiI and dye-polymer conjugates, transfer occured to a lower extent. The studies of dye release kinetics showed that the transfer of IR780 from NP to proteins occurs at a high extent (~50%) and rate, while Nile Red was slowly released from the NP over time with reduced association with proteins (~20%). This experiment assesses the stability of fluorescence labeling of nanocarriers and probes the transfer of fluorescent dyes from NP to proteins, which is otherwise not accessible with commonly used techniques of separation, such as dialysis and ultrafiltration/centrifugation employed in drug encapsulation and release studies of nanocarriers. Determining the interaction and transfer of dyes to proteins is of utmost importance in the pre-clinical evaluation of drug nanocarriers for improved correlation between in vitro and in vivo studies.
Assuntos
Proteínas Sanguíneas/análise , Portadores de Fármacos/química , Corantes Fluorescentes/química , Fracionamento por Campo e Fluxo/métodos , Nanopartículas/química , Polímeros/química , Adsorção , Fluorescência , Humanos , Hidrodinâmica , Cinética , Oxazinas/química , Teoria Quântica , Rodaminas/química , Espalhamento de RadiaçãoRESUMO
In this study, molecular properties of wheat starch from three different types of breads were analyzed using asymmetric flow field-flow (AF4) connected to multi-angle light scattering (MALS) and differential refractive index (dRI) detectors. This analysis allowed the determination of molecular properties, i.e. molar mass (M), root-mean-square radius (rrms), apparent density (ρapp) and conformation. Complementary analyses, such as resistant starch and amylose content, were also performed. The results show that wheat starch extracted from breads can have different properties reflected in changes in M, rrms and ρapp. In addition, the results suggest that some of the changes in molecular properties may be related to the presence of resistant starch.
Assuntos
Pão/análise , Fracionamento por Campo e Fluxo/métodos , Refratometria/métodos , Amido/química , Triticum/química , Amilose/análise , Bolívia , Análise de Alimentos/métodos , Peso Molecular , Espalhamento de RadiaçãoRESUMO
Yacon (Smallanthus sonchifolius Poepp. & Endl.) roots are largely grown in Andean countries and have attracted recent interest due to their antioxidant and prebiotic effects. Yacon is typically consumed as a fruit due to its sweet taste and juiciness. The macromolecular properties of an aqueous extract of yacon are investigated using asymmetric flow field-flow fractionation (AF4) coupled to UV, multiangle light scattering (MALS) and differential refractive index (dRI) detection. The method allows for determination of molar mass and size over the size distribution. Three major populations were found of which one strongly dominates in concentration. Through collection of fractions from AF4, carbohydrate composition and glycosidic linkage analysis for the dominating population was performed. The results show that the dominating population consists of a highly branched arabinogalactan (type 2) with a molar mass of approximately 1-2·105g/mol, a hydrodynamic radius of approximately 6-10nm and a relatively high apparent density (approx. 70-150kg/m3).