In this work, a liquid crystal (LC) biosensor based on whispering gallery mode (WGM) lasing is reported and demonstrated for real-time and high-sensitive detection of acetylcholinesterase (AChE) and its inhibitors. Benefit from the double amplification by the WGM resonance and the 5CB molecules, small changes in biological processes are significantly enhanced in WGM spectral responses. The spectral responses provide direct information about the molecular adsorption/desorption at the LC/aqueous solution interface and can be used as an indicator of the enzymatic reaction. The limit of detection achieved was as low as 0.1 pg/mL for fenobucarb and 1 pg/mL for dimethoate, which is considerably lower than the standard levels of pesticides specified for water quality standards. Results indicate that this versatile method holds prospects for application in real-time and high-sensitive monitoring of biochemical reactions and can serve as an alternative solution to compensate for missing capabilities in conventional POM observations.
In this study, porous sandwich structure Fe 3 O 4 nanoparticles coated by polyhedral oligomeric silsesquioxanes and β-cyclodextrin were prepared by surface polymerization and were used as the magnetic solid phase extraction adsorbent for the extraction and determination of carbaryl and carbofuran. The Fe 3 O 4 nanoparticles coated with polyhedral oligomeric silsesquioxanes and β-cyclodextrin were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, vibrating sample magnetometry, and scanning electron microscopy. After optimizing the extraction conditions, a method that combined magnetic solid phase extraction with high-performance liquid chromatography was developed for the determination of carbaryl and carbofuran in apple. The method exhibited a good linearity in the range of 2–400 μg/kg for carbaryl and carbofuran ( R 2 = 0.9995), respectively. The limits of detection were 0.5 μg/kg of carbaryl and 0.7 μg/kg for carbofuran in apple, respectively. Extraction recoveries ranged from 94.2 to 103.1% with the preconcentration factor of 300 and the relative standard deviations were less than 5.9%. These results indicated that the method combined magnetic solid phase extraction with high-performance liquid chromatography and was promising for the determination of carbaryl and carbofuran at trace amounts.
Yidong Zhou,Zhijian Ke,Hangting Ye,Mengting Hong,Yifei Xu,Mingliang Zhang,Wankui Jiang,Qing Hong
Carbaryl is a widely used carbamate pesticide in agriculture. The strain Rhizobium sp. X9 possesses the typical carbaryl degradation pathway in which carbaryl is mineralized via 1-naphthol, salicylate, and gentisate. In this study, we cloned a carbaryl hydrolase gene cehA and a novel two-component 1-naphthol hydroxylase gene cehC1C2. CehA mediates carbaryl hydrolysis to 1-naphthol and CehC1, an FMNH2 or FADH2-dependent monooxygenase belonging to the HpaB superfamily, and hydroxylates 1-naphthol in the presence of reduced nicotinamide-adenine dinucleotide (FMN)/flavin adenine dinucleotide (FAD), and the reductase CehC2. CehC1 has the highest amino acid similarity (58%) with the oxygenase component of a two-component 4-nitrophenol 2-monooxygenase, while CehC2 has the highest amino acid similarity (46%) with its reductase component. CehC1C2 could utilize both FAD and FMN as the cofactor during the hydroxylation, although higher catalytic activity was observed with FAD as the cofactor. The optimal molar ratio of CehC1 to CehC2 was 2:1. The Km and Kcat/Km values of CehC1 for 1-naphthol were 74.71 ± 16.07 μM and (8.29 ± 2.44) × 10–4 s–1·μM–1, respectively. Moreover, the enzyme activities and substrate spectrum between CehC1C2 and previously reported 1-naphthol hydroxylase McbC were compared. The results suggested that McbC had a higher 1-naphthol hydroxylation activity, while CehC1C2 had a broader substrate spectrum.
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