All reagents were analytical grade or higher. Five standards of neonicotinoid insecticide including clothianidin, imidacloprid, acetamiprid, and thiamethoxam were obtained from Dr. Ehren-storfer GmbH (Augsburg, Germany), and thiacloprid were purchased from Sigma-Aldrich (Taufkirchen, Germany). All standard neonicotinoid insecticides were of analytical standard grade. The stock solution of each insecticide (1000 µg mL−1) was prepared by dissolving an appropriate amount in methanol. Working standard solutions were prepared by diluting the stock standard solution with water. Chromatographic grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). Sodium chloride (NaCl), anhydrous sodium sulphate (anh. Na2SO4), sodium carbonate (Na2CO3) and sodium hydroxide (NaOH) were obtained from Ajax Finechem (Auckland, New Zealand), Sodium acetate (CH3COONa) was obtained from Carlo Erba (Val de Reuil, France). Triton™ X-114 was obtained from Sigma-Aldrich (St. Louis, MI, USA). Sodium dodecyl sulfate (SDS) and Triton™ X-100 were provided by Merck. Cetyltrimethyl ammonium bromide (Calbiochem, Germany) was also used. Water used throughout the experiment was produced by RiOsTM Type I Simplicity 185 (Millipore Waters, Burlington, MA, USA) with the resistivity of 18.2 MΩ.cm.
Clothianidin
Manufactured by LGC
Sourced in Germany
Clothianidin is a neonicotinoid insecticide used in the production of various agricultural and horticultural products. It functions as a systemic insecticide, which means it is absorbed by the plant and distributed throughout its tissues. The core function of Clothianidin is to provide protection against a wide range of sucking and chewing insect pests.
Automatically generated - may contain errors
Lab products found in correlation
Imidacloprid, by LGC (3 mentions)
Acetamiprid, by LGC (2 mentions)
Thiamethoxam, by LGC (2 mentions)
Triton x 114, by Merck Group (1 mentions)
Thiacloprid, by Merck Group (1 mentions)
Sodium acetate ch3coona, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions)
Ovalbumin (ova), by Merck Group (1 mentions)
Dinotefuran, by LGC (1 mentions)
Sodium sulfate, by Merck Group (1 mentions)
Milli q system, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
4 protocols using clothianidin
1
Neonicotinoid Insecticide Analysis
2
Anti-Imidacloprid Monoclonal Antibody Production
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The anti-imidacloprid monoclonal antibody was produced in the State & Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China [37 (link)]. Acetamiprid, thiacloprid, thiamethoxam, imidacloprid, dinotefuran, nitenpyram, clothianidin and 2,4-D were purchased from Dr. Ehrenstorfer (Augsburg, Germany). Horseradish peroxidase (HRP) Conjugated Streptavidin was purchased from Boster Biological Technology Co., Ltd. (Wuhan, China). (+)-biotin N-hydroxysuccinimide ester (BNHS), polyoxyethylene sorbitan monolaurate (Tween-20), 3,3′,5,5′-tetramethylbenzidine (TMB), and Ovalbumin (OVA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Chemical reagents such as 3-mercaptopropionic acid (3-MPA), dimethyl sulfoxide (DMSO), H2O2, and others were all purchased from Aladdin (Shanghai, China).
3
Bee Exposure to Clothianidin and Fungicides
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Clothianidin (purity 99%) was purchased from Dr Ehrenstorfer Gmbh. The stock solution with a concentration of 1.05 mg of Clothianidin/mL of acetone was used to prepare the test solutions. To obtain a range of appropriate concentrations based on the desired exposure level, the stock solution was first diluted in acetone until we reached the final nominal concentrations ranging from 2 to 160 mg/L (actual concentrations: 2.7 to 176.8 mg/L). These solutions were added to the feeding solution (500 g of sucrose in 1 L of purified distilled water) at the ratio of 10 µL/mL (see 2.3 Experimental design).
The final concentration of acetone in the feeding solution was adjusted to 3% (v:v) in all treatments (solvent control, propiconazole, Clothianidin and the combination fungicideneonicotinoid). To reach this level, pure acetone was added as needed. Following the exposure phase, bees were fed ad libitum with a sucrose solution (33% w:w).
The final concentration of acetone in the feeding solution was adjusted to 3% (v:v) in all treatments (solvent control, propiconazole, Clothianidin and the combination fungicideneonicotinoid). To reach this level, pure acetone was added as needed. Following the exposure phase, bees were fed ad libitum with a sucrose solution (33% w:w).
4
Purity Quantification of Neonicotinoid Pesticides
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Imidacloprid, clothianidin, and thiamethoxam were purchased from Dr. Ehrenstorfer GmbH (Augsburg, Germany). Purity assays of these pesticides were performed according to protocols developed in this laboratory at the National Metrology Institute of Korea. Details of the purity assay have been described in previous studies (Kim et al. 2013; Lee and Kim 2014) . Briefly, LC/UV analysis for structure-related impurities, Karl-Fisher titration for water content, thermogravimetric analysis for nonvolatile impurities, and head-space GC/MS for residual solvents were performed, respectively, and the chemicals were subsequently used without further purification. The purity of Imidacloprid, clothianidin, and thiamethoxam was determined as (99.5 ± 0.3)%, (99.4 ± 0.3)%, and (99.6 ± 0.1)%, respectively. Isotope analogues, Imidacloprid-d 4 , clothianidin-d 3 , and thiamethoxam-d 4 , were obtained from LGC (Teddington, UK). Ammonium acetate for the aqueous mobile phase was purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium chloride and sodium sulfate for sample preparation procedure were also purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade organic solvents, methanol, and acetonitrile were obtained from Burdick and Jackson (Musekegon, MI, USA). Water was purified with a Millipore Milli-Q system at 18.2 MΩ/cm 2 .
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