Competitive immune-nanoplatforms with positive readout for the rapid detection of imidacloprid using gold nanoparticles.

Two high-sensitivity competitive immune-nanoplatforms based on the inner filter effect (IFE-IN) and magnetic separation (MS-IN) with a positive readout were developed to rapidly detect imidacloprid (IMI) using gold nanoparticles (AuNPs). For IFE-IN, IMI competes with AuNPs-labeled IMI antigens (IMI-BSA-AuNPs) to bind with anti-IMI monoclonal antibody (mAb)-conjugated NaYF4:Yb,Er upconversion nanoparticles, which changes the fluorescence signal at excitation/emission wavelength of 980/544 nm. For MS-IN, the immunocomplex of IMI-BSA-AuNPs and magnetic-nanoparticles-labeled mAb (mAb-MNPs) dissociates in the presence of IMI, and the optical density of IMI-BSA-AuNPs at 525 nm increases with the IMI concentration after magnetic separation. Under the optimal conditions, the IMI concentration producing a 50% saturation of the signal (SC50) and linear range (SC10- SC90) were found to be 4.30 ng mL-1 and 0.47 - 21.37 ng mL-1 for IFE-IN, while 1.21 ng mL-1 and 0.07 - 10.21 ng mL-1 for MS-IN, respectively. Both IFE-IN and MS-IN achieved excellent accuracy for the detection of IMI in different matrices. The quantities of IMI in apple samples detected by IFE-IN and MS-IN were consistent with the high-performance liquid chromatography results. For IFE-IN, analyte competes with AuNPs-labeled-antigen to bind with the mAb-conjugated-UCNPs, which changes the fluorescence signal at 544 nm. For MS-IN, the immunocomplex of AuNPs-labeled-antigen and mAb-conjugated-MNPs dissociates in the presence of analyte, and the optical density of AuNPs-labeled-antigen at 525 nm increases with increasing analyte concentration after separation.

Upconversion fluorescence immunoassay for imidaclothiz by magnetic nanoparticle separation.

A sensitive fluorescence immunoassay for the detection of imidaclothiz was established by using magnetic nanoparticles (MNPs) as concentration elements and upconversion nanoparticles (UCNPs) as signal labels. The NaYF4/Yb,Er UCNPs and MNPs were conjugated with imidaclothiz monoclonal antibody and imidaclothiz antigen, respectively. Imidaclothiz could compete with the antigen-conjugated MNPs for binding to the antibody-conjugated UCNPs and resulted in a decreased fluorescence signal when the MNPs were separated by an external magnet. Under the optimal conditions, the concentration of imidaclothiz producing 50% inhibition of the signal (IC50), limit of detection (LOD, IC10), and the linear assay range (IC10-IC90) were 14.59, 0.74, and 0.74-289.30 ng mL-1, respectively. The immunoassay exhibited no obvious cross-reactivity with analogues of imidaclothiz except for imidacloprid, with 89.2% cross-reactivity. The average recoveries measured in paddy water, pear, soil, peach, rice, tomato, wheat, and pakchoi were 75.7-105.2%, and the relative standard deviations (RSDs) were less than 11.2%. In addition, the results of the immunoassay correlated well with that of high-performance liquid chromatography (HPLC) for authentic samples. Graphical abstract Development of an upconversion fluorescence immunoassay for the detection of imidaclothiz by using antibody-modified upconversion nanoparticles (UCNPs) as the detection probe and antigen-modified magnetic nanoparticles (MNPs) as the capture probe.

Development of Fluorescence Polarization Immunoassay for Imidacloprid in Environmental and Agricultural Samples.

A fluorescence polarization immunoassay (FPIA) for the determination of imidacloprid (IMI) was developed with advantages of simple operation and short assay time. The haptens of IMI, acetamiprid (ACE), and thiamethoxam (THI) were conjugated with fluorescein isothiocyanate ethylenediamine (EDF) and 4'-Aminomethyl fluorescein (AMF), respectively, to prepare six fluorescence tracers. The conjugation of IMI hapten and EDF (IMI-EDF) was selected to develop the FPIA due to the largest fluorescent polarization value increase in the presence of anti-IMI monoclonal antibody. Under the optimum condition, the limit of detection, 50% inhibition concentration and detection range of the FPIA were 1.7, 4.8, and 1.7-16.3 µg/L, respectively. The cross-reactivities (CRs) with the analogs of IMI were negligible except for imidaclothiz with CR of 79.13%. The average recovery of spiked paddy water, corn and cucumber samples were 82.4-118.5% with the RSDs of 7.0-15.9%, which indicated the FPIA had good accuracy. Thus, the developed FPIA was a potential tool for the rapid and accurate determination of IMI in agricultural and environmental samples.

Competitive immunoassay for simultaneous detection of imidacloprid and thiacloprid by upconversion nanoparticles and magnetic nanoparticles.

A rapid and sensitive immunoassay for the simultaneous detection of imidacloprid and thiacloprid was developed by using magnetic nanoparticles (MNPs) and upconversion nanoparticles (UCNPs). The UCNPs of NaYF4:Yb, Er and NaYF4:Yb, Tm were synthesized and conjugated with anti-imidacloprid monoclonal antibody (mAb) and anti-thiacloprid mAb as signal labels, while the MNPs were conjugated with antigens of thiacloprid and imidacloprid as separation elements. The fluorescence intensities of Yb/Er- and Yb/Tm-doped UCNPs were detected simultaneously in 544 nm and 477 nm under the excitation of NIR light (980 nm). The amounts of mAb-conjugated UCNPs that were separated by antigen-conjugated MNPs were determined based on competitive immunoassays. Under the optimal conditions, the 50% inhibiting concentration (IC50) and limit of detection (LOD, IC10) were 5.80 and 0.32 ng/mL for imidacloprid and 6.45 and 0.61 ng/mL for thiacloprid, respectively. The immunoassay exhibited negligible cross-reactivity with analogs of imidacloprid and thiacloprid except imidaclothiz (86.2%). The average recoveries of imidacloprid and thiacloprid in environmental and agricultural samples, including paddy water, soil, pears, oranges, cucumbers, and wheat, ranged from 78.4 to 105.9% with relative standard deviations (RSDs) of 2.1-11.9% for imidacloprid and ranged from 82.5 to 102.3% with RSDs of 1.0-16.5% for thiacloprid. In addition, the results of the immunoassay correlated well with high-performance liquid chromatography for the detection of the authentic samples.

Development of immunochromatographic assays for the detection of imidacloprid in soil chemical barrier.

A soil chemical barrier is the most important and common way to control termites; fast and on-site detection methods are significant tools to verify pesticide content meeting the standard requirements. In this study, conventional and enhanced immunochromatographic assays (ICAs) containing two test lines (TLs) were developed to semi-quantitatively detect imidacloprid in soil chemical barrier, and detection results were quantified by a smart phone. According to the results, the disappearance concentrations of first TL (TL-1) and second TL (TL-2) in an enhanced ICA and conventional ICA were 5 and 20 ng/mL and 20 and 80 ng/mL with the naked eye. The sensitivity of TL-2 was four times that of TL-1 in both ICAs, consistent with the maximum and minimum concentration differences for imidacloprid in Jiangsu province's "the technical regulation of assay and evaluation on chemical soil barrier of termite prevention treatment in buildings". The results of TLs can be used to judge whether the amount of imidacloprid in soil chemical barrier meets the standard. Enhanced and conventional ICAs were available for further quantitative testing with a smart phone, and the limit of detection (LOD) was 0.74 and 3.17 ng/mL, respectively. Moreover, some soil chemical barrier samples from several areas in Wuxi, Jiangsu province, were used to test by ICAs and high-performance liquid chromatography (HPLC), and the results of ICAs correlated well with HPLC.

Development of an upconversion fluorescence DNA probe for the detection of acetamiprid by magnetic nanoparticles separation.

An upconversion fluorescence DNA probe which consists of aptamer-conjugated magnet nanoparticles (apt-MNPs) and complementary DNA-conjugated upconversion nanoparticles (cDNA-UCNPs) was developed to detect acetamiprid. Acetamiprid can specifically conjugate with the apt-MNPs to dissociate the cDNA-UCNPs from the apt-MNPs and resulted in reduced fluorescence intensity through an external magnet. The change of fluorescence intensity (△I) is positively related to the concentration of acetamiprid, which can be applied for the quantification of acetamiprid. Under optimal conditions, a linear detection range and detection limit are 0.89-114.18 µg/L and 0.65 µg/L, respectively. The probe was successfully used to detect acetamiprid in spiked paddy water, soil, pear, apple, wheat and cucumber. Average recoveries are 78.2%-103.5% with intra-day relative standard deviations (RSDs) of 2.6%-10.9% and inter-day RSDs of 4.3%-10.2%. The amounts of acetamiprid in the authentic paddy water and pear samples detected by the DNA probe are significantly correlated with that detected by high-performance liquid chromatography (HPLC).