A simple uric acid assay by using 3-hydroxytyramine as a chromogenic colorimetric sensor in human serum samples: Density functional theory supported mechanistic approach

A simple, fast, sensitive, convenient, and economical accurate enzymatic colorimetric sensor is developed for the quantification of the clinically important biomarker uric acid (UA) in human serum samples. The method is based on the quantification of UA using 3-hydroxytyramine (3-HT) as a chromogenic substrate by using peroxidase (POD) and uricase (UOx). UOx catalyzes the oxidation of UA where H₂O₂ is generated in-situ to produce allantoin and CO₂. The POD in oxidative coupling of peroxide in presence of 3-HT, forms an intense orange-colored product with λ_max = 500 nm. The reaction was carried out in a citric acid-tripotassium-citrate buffer (12.5 mM) of pH = 6.8 at room temperature. Computational calculations were done by Gaussian-16 employing the B3LYP functional basis set. The standard curve for UA was found to be linear between 12.3–297.3 and 3.07–396.5 μM by rate and fixed time method, respectively. The LOD and LOQ for UA were 1.5 and 2.9 μM, respectively. Within-day and day-to-day precision was 1.50–3.07 and 3.10–4.16% (n = 7), respectively. The accuracy ranges for UA concentrations of 24.7, 198 and 297.3 μM were 87%–101.50%, 90%–105%, and 89%–107%, respectively. The UA recovery range by the proposed method was 98.00%–100.47% with a mean recovery of 99.5% and has A good correlation coefficient of 0.981 with the standard method.     

Synthetic Monosaccharide Channels: Size-Selective Transmembrane Transport of Glucose and Fructose Mediated by Porphyrin Boxes

Here we report synthetic monosaccharide channels built with shape-persistent organic cages, porphyrin boxes ( PB s), that allow facile transmembrane transport of glucose and fructose through their windows. PB s show a much higher transport rate for glucose and fructose over disaccharides such as sucrose, as evidenced by intravesicular enzyme assays and molecular dynamics simulations. The transport rate can be modulated by changing the length of the alkyl chains decorating the cage windows. Insertion of a linear pillar ligand into the cavity of PB s blocks the monosaccharide transport. In vitro cell experiment shows that PB s transport glucose across the living-cell membrane and enhance cell viability when the natural glucose transporter GLUT1 is blocked. Time-dependent live-cell imaging and MTT assays confirm the cyto-compatibility of PB s. The monosaccharide-selective transport ability of PB s is reminiscent of natural glucose transporters (GLUTs), which are crucial for numerous biological functions.