Yujuan Cao

Molecularly imprinted polymer coated solid-phase microextraction fiber prepared by surface reversible addition-fragmentation chain transfer polymerization for monitoring of Sudan dyes in chilli tomato sauce and chilli pepper samples.

Surface reversible addition-fragmentation chain transfer (RAFT) polymerization method was firstly applied to the preparation of molecularly imprinted polymer (MIP) coated silicon solid-phase microextraction (SPME) fibers. With Sudan I as template, an ultra-thin MIP coating with about 0.55-μm thickness was obtained with homogeneous structure and controlled composition, due to the controllable radical growing and chain propagation in surface RAFT polymerization. The MIP-coated fibers were found with enhanced selectivity coefficients (3.0-6.5) to Sudan I-IV dyes in contrast with those reported in our previous work. Furthermore, the ultra-thin thickness of MIP coating was helpful to the effective elution of template and fast adsorption/desorption kinetics, so only about 18 min was needed for MIP-coated SPME operation. The detection limits of 21-55 ng L(-1) were achieved for four Sudan dyes, when MIP-coated SPME was coupled with liquid chromatography (LC) and mass spectrometry (MS) detection. The MIP-coated SPME-LC-MS/MS method was tested for the monitoring of ultra trace Sudan dyes in spiked chilli tomato sauce and chilli pepper samples, and high enrichment effect, remarkable matrix peaks-removing capability, and consequent high sensitivities were achieved to four Sudan dyes.

Molecularly imprinted polymer coated solid-phase microextraction fibers for determination of Sudan I–IV dyes in hot chili powder and poultry feed samples

In this research, a novel strategy was developed to prepare molecularly imprinted polymer (MIP) coated solid-phase microextraction fibers on a large scale with Sudan I as template and stainless steel fibers as substrate. More than 20 fibers could be obtained in one glass tube, and the efficiency and coating repeatability were enhanced remarkably in contrast with the yield of only one fiber in our previous works. The obtained MIP-coated stainless steel fibers were characterized by homogeneous and highly cross-linked coating, good chemical and thermal stabilities, high extraction capacities, and specific selectivities to Sudan I-IV dyes. Based on the systemic optimization of extraction conditions, a simple and cost-effective method based on the coupling of MIP-coated SPME with high-performance liquid chromatography was developed for the fast and selective determination of trace Sudan I-IV dyes in hot chili powder and poultry feed samples. The limits of detection of Sudan I-IV dyes were within 2.5-4.6 ng g(-1), and the spiked recoveries were in the range of 86.3-96.3% for hot chili powder sample and 84.6-97.4% for poultry feed sample.

Novel liquid–liquid–solid microextraction method with molecularly imprinted polymer-coated stainless steel fiber for aqueous sample pretreatment

A novel liquid-liquid-solid microextraction (LLSME) method was developed to overcome the well-known water-compatibility problem of molecularly imprinted polymers (MIPs). The enrichment factors with MIP-LLSME method were within 70-210 for trace chloroacetanilide herbicides under optimized extraction conditions. The method was characterized by simplicity, low solvent-consumption and high selectivity, and it was suitable for the one-step pretreatment of various aqueous samples such as river water and farm water.

A Highly Selective and Sensitive Fluorescence Detection Method of Glyphosate Based on an Immune Reaction Strategy of Carbon Dot Labeled Antibody and Antigen Magnetic Beads

A sensitive fluorescence detection method for glyphosate (GLY) was established based on immune reaction. First, carbon dot labeled antibodies (lgG-CDs) which were able to specifically identify glyphosate were prepared with the environmentally friendly carbon dots (CDs) and glyphosate antibody (lgG). lgG-CDs could be used to in situ visualize the distribution of glyphosate in plant tissues. In order to eliminate the effects of excess lgG-CDs on the determination of GLY, antigen magnetic beads Fe3O4-GLY based on magnetic nanoparticles Fe3O4 and glyphosate were constructed and utilized to couple with the excess lgG-CDs. After magnetic separation to remove antigen magnetic beads, there was a linear relationship between the fluorescence intensity of lgG-CDs and the logarithmic concentration of glyphosate in the range of 0.01-80 μg/mL with a detection limit of 8 ng/mL. The method was used for the detection of glyphosate in Pearl River water, tea, and soil samples with satisfactory recovery ratio between 87.4% and 103.7%.

Recognition of DNA based on changes in the fluorescence intensity of CdSe/CD QDs–phenanthroline systems

The CdSe quantum dots (QDs) modified by mercapto-beta-cyclodextrin (CD) were synthesized and characterized by transmission electron microscopy, powder X-ray diffraction, excitation and emission spectra, and fluorescence lifetime. When lambda(ex)=370nm, the fluorescence peak of CdSe/CD QDs is at 525nm. Phenanthroline (Phen) is able to quench their fluorescence, which can be recovered by the addition of DNA. The quenching and restoration of fluorescence intensity were found to be linearly proportional to the amount of Phen and DNA, respectively. The variation of the fluorescence intensity of the CdSe/CD QDs-Phen system was studied, and it was demonstrated to result from a static mechanism due to the formation of a Phen inclusion complex with the CdSe QDs modified by mercapto-beta-cyclodextrin. The fluorescence recovery was due to the binding of DNA with Phen in the inclusion complex, leading to the freeing of the CdSe/CD QDs. The binding constants and sizes of the binding sites of the Phen-DNA interaction were calculated to be 1.33x10(7)mol(-1)L and 10.79bp.

Synthesis and application of intercellular Ca2+-sensitive fluorescent probe based on quantum dots.

A novel Ca(2+)-sensitive fluorescent probe was synthesized and characterized with a coupled method that coupled di[2-(N,N-dicarboxylmethyl)amino]ethyl ether (EGTA) to the surface of mercaptoethylamine-modified CdTe quantum dots (CdTe/MA-EGTA QDs). The application of this probe to detect intercellular Ca(2+) change in the leaf cells of Arabidopsis thaliana was studied. Results from transmission electron micrographs showed that the particle size of CdTe/MA-EGTA was about 3-4 nm; the fluorescent spectrum indicated that the excitation spectral ranged from 350 to 490 nm with a narrow and symmetric emission spectral peak at 565 nm when excited by 400 nm, and capillary electrophoresis demonstrated that CdTe/MA-EGTA was obtained by a coupling reaction. When the detected conditions were set as an excitation wavelength of 514 nm and detection wavelength of 561-604 nm, the increase of Ca(2+) in A. thaliana leaf cells and the rapidly quenching effect of fluorescence signal induced by exogenous treatment of jasmonate acid (JA) could be measured using laser scanning confocal microscopy. The quenching rate of traditional Ca(2+)-sensitive fluorescent probe Fluo-3 reached about 80% within a minute when exciting at 488 nm, which was much faster than the novel fluorescent probe CdTe/MA-EGTA. CdTe/MA-EGTA, however, was better at resisting photo bleaching and was more suitable for long-term tracking and monitoring than Fluo-3.

Point-of-care testing for streptomycin based on aptamer recognizing and digital image colorimetry by smartphone

The rapid detection of antibiotic residual in everyday life is very important for food safety. In order to realize the on-site and visual detection of antibiotic, a POCT method was established by using digital image colorimetry based on smartphone. Streptomycin was taken as the analyte model of antibiotics, streptomycin aptamer preferentially recognized analyte, and the excess aptamer hybridized with the complementary DNA to form the dsDNA. SYBR Green I combined with the dsDNA and then emitted obvious green fluorescence, thus the fluorescence intensity decreased with the increasing of streptomycin concentration. Then a smartphone-based device was constructed as the fluorescence readout. The smartphone camera acquired the images of the fluorescence derived from the samples, and the Touch Color APP installed in smartphone read out the RGB values of the images. There was a linear relationship between the G values and the streptomycin concentrations in the range of 0.1-100µM. The detection limit was 94nM, which was lower than the maximum residue limit defined by World Health Organization. The POCT method was applied for determining streptomycin in chicken and milk samples with recoveries in 94.1-110%. This method had the advantages of good selectivity, simple operation and on-site visualization.

In situ detection of salicylic acid binding sites in plant tissues

The determination of hormone-binding sites in plants is essential in understanding the mechanisms behind hormone function. Salicylic acid (SA) is an important plant hormone that regulates responses to biotic and abiotic stresses. In order to label SA-binding sites in plant tissues, a quantum dots (QDs) probe functionalized with a SA moiety was successfully synthesized by coupling CdSe QDs capped with 3-mercaptopropionic acid (MPA) to 4-amino-2-hydroxybenzoic acid (PAS), using 1-ethyl-3-(3-dimethyllaminopropyl) carbodiimide (EDC) as the coupling agent. The probe was then characterized by dynamic light scattering and transmission electron microscopy, as well as UV/vis and fluorescence spectrophotometry. The results confirmed the successful conjugation of PAS to CdSe QDs and revealed that the conjugates maintained the properties of the original QDs, with small core diameters and adequate dispersal in solution. The PAS-CdSe QDs were used to detect SA-binding sites in mung bean and Arabidopsis thaliana seedlings in vitro and in vivo. The PAS-CdSe QDs were effectively transported into plant tissues and specifically bound to SA receptors in vivo. In addition, the effects of the PAS-CdSe QDs on cytosolic Ca(2+) levels in the tips of A. thaliana seedlings were investigated. Both SA and PAS-CdSe QDs had similar effects on the trend in cytosolic-free Ca(2+) concentrations, suggesting that the PAS-CdSe QDs maintained the bioactivity of SA. To summarize, PAS-CdSe QDs have high potential as a fluorescent probe for the in vitro/in vivo labeling and imaging of SA receptors in plants.

Fluorescence Determination of Omethoate Based on a Dual Strategy for Improving Sensitivity

Omethoate is a frequently used organophosphorus pesticide, and the establishment of a sensitive, selective, and simple method to determine omethoate is very important for food safety. In this paper, a dual strategy was applied to improve the detection sensitivity of omethoate. In the first strategy, graphene quantum dots (GQDs) were doped with nitrogen to increase the fluorescence quantum yield to 30%. By coupling N-GQDs with omethoate aptamer, an N-GQDs-aptamer probe was synthesized. The fluorescence of the N-GQDs-aptamer probe was turned off by graphene oxide (GO), but recovered by omethoate. Based on this principle, the fluorescence method for detecting omethoate was established with a detection limit of 0.041 nM. To further improve the detection sensitivity, the fluorescence polarization analysis method was applied as another strategy based on the polarization signal of GQDs. The detection limit was decreased to 0.029 pM by using the fluorescence polarization method. The detection limits in this paper were lower than those in other reports. The imaging of omethoate on plant leaves showed that the probe could be used for visual semiquantitative determination of omethoate.

In situ fluorescence labelling of jasmonic acid binding sites in plant tissues with cadmium-free quantum dots.

The fluorescence labelling of plant hormone binding sites is an important analytical technique in research on the molecular mechanisms of plant hormone activities. The authors synthesised a jasmonic acid (JA)-conjugated ZnS:Mn quantum dot (QD) probe, with a cubic structure and average hydrodynamic sizes of about 17.0 nm. The maximum fluorescence emission of the probe was recorded at about 585 nm. The probe was used for fluorescence labelling of JA binding sites in mung bean seedling tissues. Analysis revealed that the probe exhibited high selectivity to JA binding sites and good performance in eliminating interference from background fluorescence in plant tissues. In addition, the probe did not exhibit any apparent biotoxicity, and is much more suitable than probes constructed from CdTe QDs for the analysis of biological samples.