In this work, a novel approach for simple and sensitive determination of alkaline phosphatase (ALP) is developed on the basis of an inner filter effect of p-nitrophenylphosphate (PNPP) on the fluorescence of gold nanoclusters (AuNCs). AuNCs with a high quantum yield of 12% were synthesized by one-pot strategy and were directly applied as fluorescent substance. When AuNCs were mixed with PNPP, the fluorescence of the AuNCs was remarkably quenched or was decreased via the inner filter effect since the absorption spectrum of PNPP overlaps well with the excitation spectrum of the AuNCs. While in the presence of ALP, PNPP was catalytically hydrolyzed into p-nitrophenol, which has different absorption characteristics from those of PNPP, resulting in the recovery of the AuNCs fluorescence. Thus, a novel "turn-on" fluorescent sensor for detecting ALP was established with a detection limit as low as 0.002 U/L (signal-to-noise ratio of 3). The turn-on fluorescent sensor exhibits many merits such as high sensitivity, excellent selectivity, and high signal output because of the low background signals. In addition, the developed sensing method was successfully applied to investigate ALP inhibitors and ALP determination in serum samples. A good linear relationship was obtained in the range from 0.02 to 50 U/L, and satisfactory recoveries at four spiking levels of ALP ranged from 95% to 106% with precision below 5%. The very simple sensing approach proposed here should promote the development of fluorescence turn-on chemosensors for chemo/biodetection.
Release assessment of organo-modified montmorillonite (O-MMT) nanoclay and the organo-modifiers (surfactants) was performed on two types of polymer–clay nanocomposites: polypropylene (PP) and polyamide 6 (PA6) with O-MMT. In accordance with ASTM D4754-11, nanocomposite films were exposed to ethanol as a fatty-food simulant at 70 °C. The release of O-MMT, with Si and Al used as the nanoclay markers, was evaluated by graphite furnace atomic absorption spectrometry. The nanoclay particles released in ethanol were visualized by transmission electron microscopy (TEM). More nanoclay particles were released from PP–clay films (0.15 mg L(–1)) than from PA6–clay films (0.10 mg L(–1)), possibly due to the lack of interaction between the nanoclay and PP as indicated by the structure and morphology in the TEM images. The surfactant release was quantified by a liquid chromatography tandem mass spectrometry (LC-MS/MS) method. A substantial amount of surfactant was released into ethanol (3.5 mg L(–1) from PP–clay films and 16.2 mg L(–1) from PA6–clay films), indicating changes in the nanoclay structure within the nanocomposite while it was exposed to ethanol. This research has provided information for the determination of exposure doses of nanoclay and surfactant in biosystems and the environment, which enabled the risk assessment.
We report a methodology developed to detect and track stable fluorescent-labeled nanoclay, in polymer-clay nanocomposite films, and in a contact solvent after migration testing. Fluorescein-5-maleimide (fluorescein) or tetramethylrhodamine-5-maleimide (rhodamine) was covalently bonded to organically modified montmorillonite (o-MMT). Fluorescein- and rhodamine-labeled nanoclay showed good thermal stability up to 220 °C and the rhodamine-labeled nanoclay remained stable at 250 °C. Confocal laser scanning microscopy was used to confirm the tagging and to detect the fluorescent-labeled nanoclays in various systems.
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