Monoclonal antibody-based enzyme-linked immunosorbent assay for the insecticide imidacloprid

An enzyme-linked immunosorbent assay (ELISA) was developed for the neonicotinoid insecticide imidacloprid, 1-[(6-chloro-3-pyridinyl)methyl]-N-nitro-2-imidazolidinimine using monoclonal antibodies (MAb). Three MAbs, designated as E6A6, E6F3 and H7F7, were raised from mice immunized with an imidacloprid hapten-ovalbumin conjugate. These MAbs performed similarly in indirect competition ELISA (icELISA), so one, E6F3, was selected for detailed study. The equilibrium constants (K_d) and association and dissociation rate constants (k_on, k_off) for five neonicotinoids and one imidacloprid metabolite to E6F3 were determined by kinetic exclusion fluoroimmunoassay (KinExA). Affinities (1/K_d) of E6F3 for acetamiprid and clothianidin were similar, but 50-fold weaker than that of imidacloprid. MAb E6F3 had no measurable affinity for the other neonicotinoids. The icELISA can tolerate up to 15% (v/v) acetone or 20% (v/v) methanol. Assay sensitivity was similar at pH 4-9, 1-10-fold concentration of PBS with or without 0.05% Tween 20, and incubation times of 30-180 min. The half-maximal inhibition and the limit of detection were approximately 0.8 and 0.1 μg/l of imidacloprid in icELISA, and 0.3 and 0.03 μg/l in direct competition ELISA (dcELISA), respectively. Analysis of imidacloprid-fortified water and cucumber samples by the icELISA showed average recoveries from 70 to 120%.     

Azopyridine-imidacloprid derivatives as photoresponsive neonicotinoids

A series of imidacloprid derivatives containing an azopyridine motif as a photoswitchable functional group were designed and synthesized. The new version of photoresponsive imidacloprid analogues showed improved solubility in comparison with their azobenzene analogues. 1.2 to 2-fold activity difference was observed for these azopyridine-imidacloprids against house fly (Musca domestica) and cowpea aphid (Aphis craccivora) upon irradiation.     

Zero‐Valent Aluminum as Reducer in Sodium Carbonate Solution for Degradation of Imidacloprid

Imidacloprid (IMI ) was reduced by zero‐valent aluminum (ZVAl ) in sodium carbonate solution and tested by high‐performance liquid chromatography (HPLC ). The IMI degradation rate was heavily dependent on Na₂CO₃ concentration and reaction time but only slightly on temperature, reaching 99.4% in 0.5 mol/L Na₂CO₃ solution with 5 g/L ZVAl in 60 min. The degradation of IMI follows first‐order kinetics. We used high‐performance liquid chromatography‐mass spectrometry (HPLC‐MS ) for analyzing the transformation products, and found that IMI disintegrated into IMI urea, IMI guanidine, IMI nitrosamine, and 1‐choloro‐5‐methcyclohea‐1,4‐diene.     

液相色谱-串联质谱法测定蜂产品中吡虫啉及其3种代谢物

建立了蜂蜜和蜂花粉中吡虫啉及其代谢物吡虫啉烯烃、吡虫啉脲、6-氯烟酸的液相色谱-串联质谱(LC-MS/MS)检测方法.在QuEChERS方法基础上,针对目标物化学性质和样品杂质情况,对提取溶液的pH值、提取次数、净化材料等参数进行了优化.最终以5％甲酸-乙腈溶液提取两次,无水MgSO4、NaCl盐析分层,提取液经增强型脂类去除材料(EMR)净化,以LC-MS/MS进行测定,基质外标法进行定量分析.结果表明,蜂蜜和蜂花粉中吡虫啉、吡虫啉烯烃、吡虫啉脲、6-氯烟酸的平均加标回收率为86.0％～111.5％, 相对标准偏差在1.7％～9.6％之间, 检出限分别为0.20, 3.50, 0.40和14.00 μg/kg,定量限分别为0.60, 11.64, 1.20和45.00 μg/kg.本方法分析速度快、灵敏度高、重现性好,适用于蜂蜜和蜂花粉中吡虫啉及其3种代谢物的快速测定.     

Absorption,transportation and distribution of imidacloprid in maize

The neonicotinoid imidacloprid is a very important insecticide in maize cultivation in China. A laboratory experiment was conducted to investigate its absorption, transportation and distribution in maize plants. Plants were exposed to an aqueous solution of imidacloprid at five concentrations (10, 20, 50, 100 and 200 μg/mL). The residues of imidacloprid in different plant parts were determined by using a quick and effective method of high-performance liquid chromatography. Results showed that the average recoveries ranged from 85.16% to 102.23%, with relative standard deviations of 1.82–4.40% at three different spiking levels in each different matrix. The half-lives of imidacloprid in hydroponic maize water were from 5.33 to 11.55 days. The concentrations in roots, stems and leaves were from 5.61 to 7.48, 1.03 to 4.03 and 0 to 30.57 μg/mL after 6 h–7 days exposed in 10 μg/mL imidacloprid aqueous solutions, respectively. Our study showed that imidacloprid was strongly absorbed by roots and mainly accumulated in leaves. Quantity ratio and bioconcentration factors (BCFs) were also used to estimate the distribution and accumulation in maize. The values of quantity ratios were the highest in the leaves while lowest in the root after 7 days treatments, with the ranking of leaves > stem > root. The BCFs were 0.63–1.66, 0.52–0.92 and 3.20–6.78 in root, stem and leaves, respectively. These results demonstrated that the exposed time and imidacloprid application concentrations were also the main factors influencing the absorption. This study enhances our understandings of the uptake and distribution of imidacloprid in maize plants.     

Liquid chromatography-mass spectrometry identification of imidacloprid photolysis products

Photolysis of imidacloprid by near ultraviolet (UV) radiation was investigated using reversed phase liquid chromatography (RPLC) and liquid chromatography-electrospray ionization/mass spectrometry (LC-ESI/MS) operated in the positive ion mode. The decomposition of imidacloprid by near UV light is first order with a half life of 10.18 h. Photo-degradation products of imidacloprid identified in this study included imidacloprid urea, imidacloprid olefin, and imidacloprid desnitro. A degradation pathway is proposed for imidacloprid from an examination of the product decay curves obtained in this monitoring study which was performed in a batch test mode.     

Determination of Imidacloprid in Tomato Grown in Greenhouse Based on Copper(II) Phthalocyanine Modified Carbon Ceramic Electrode by Differential Pulse Voltammetry

A new sol‐gel derived electrocatalytic carbon ceramic electrode was prepared by incorporating copper(II) phthalocyanine (CuPc) in a carbon ceramic network. This electrode was used as a sensitive electrochemical sensor for determination of the insecticide Imidacloprid (1‐(6‐chloro‐3‐pyridylmethyl)‐N‐nitro‐imidazolidin‐2‐ylideneamine) by differential pulse voltammetry (DPV). The resulting modified electrode exhibits a cathodic peak potential shifted positively and an increasing in cathodic peak current in comparison with unmodified electrode. The redox properties of this modified electrode at various pH values and CuPc percentage were investigated. The catalytic current obtained from differential pulse voltammetry is linearly dependent on Imidacloprid concentration over the two linear ranges of 0.67‐17 μM and 17‐93 μM with correlation coefficient of R² = 0.9999 and R² = 0.990, respectively. The detection limit for Imidacloprid was found to be 0.28 μM according to lower linear range. Possible interferences from several common pesticides were also evaluated. The inherent stability, high sensitivity, low detection limit and low cost for each preparation are advantages of this sensor. Determination of Imidacloprid in commercial formulation and residual Imidacloprid in tomato grown in greenhouse (protected cultivation) was also conducted. The results obtained from commercial formulation were completely consistent with those obtained through the standard high‐performance liquid chromatography (HPLC) method.     

QuEChERS-高效液相色谱法检测蔬菜中的吡虫啉、虫酰肼、阿维菌素和噻螨酮

将一种新的QuEChERS样品处理方法进行了提取及净化的改进,结合HPLC对蔬菜中吡虫啉、虫酰肼、阿维菌素和噻螨酮4种农药进行同时检测,取得了满意结果。以乙腈-水为流动相,梯度洗脱,269、240 nm双波长检测,流速1.0 mL/min。检出限分别为:吡虫啉0.02 mg/kg、虫酰肼0.02 mg/kg、阿维菌素0.10 mg/kg、噻螨酮0.05 mg/kg。在添加浓度为0.05~1.0 mg/kg范围内,4种化合物回收率在80%~100%之间,相对标准偏差1%~10%。方法适用于蔬菜中吡虫啉、虫酰肼、阿维菌素和噻螨酮4种农药残留量的同时检测。     

固相萃取-高效液相色谱法测定烟叶中吡虫啉农药残留量

建立了固相萃取-高校液相色谱法测定烟叶中吡虫啉农药残留量的方法。样品用乙腈提取,弗罗里硅土固相萃取柱净化.梯度洗脱方式分离,DAD二极管阵列检测器检测。吡虫啉加标回收率为87.83%—94.80%,相对标准偏差为1.64%,检出限为0.01μg/g。该方法灵敏、准确,适用烟草中吡虫啉农药残留的分析。     

离子交换固相萃取-高效液相色谱法研究吡虫啉在梨中的分布及残留动态

建立一种以固相萃取(SPE)、高效液相色谱(HPLC)为基础的测定梨果皮、果肉中吡虫啉残留量的方法。样品用乙腈进行提取,经二氯甲烷液液分配后用阳离子交换固相萃取(SCX-SPE)柱净化,高效液相色谱/二极管阵列检测法(HPLC/DAD)测定。各种添加水平(果皮:0.05、0.1、0.5、2 mg/kg;果肉:0.05、0.5、1mg/kg)的回收率为83%~103%,相对标准偏差(RSD)小于10%,方法的检出限为0.05 mg/kg。运用此方法分别对梨果皮和果肉中吡虫啉的残留动态进行研究,发现吡虫啉主要在梨果皮中残留,得到吡虫啉在果皮中的消解动态方程为:wt=w0×exp(-0.235t),半衰期是2.98 d。