Cp225d

Thirty males and thirty females emerged within 24 h of each treatment and controls were moved into 50 mL centrifuge tubes, respectively. These tubes were placed in a −20 °C freezer for 10 min to kill the adults, and were then transferred to an oven to be dried at 60 °C for 48 h. After the drying procedure, each dried fly was weighed individually using a semimicro balance (CP225D, accuracy of 0.01 mg, Sartorius, Göttingen, Germany).

The solubility data of DCBNT were measured by the dynamic method and collected by the CrystalSCAN system (E1320, HEL Ltd, UK; figure 4). The mass of DCBNT was weighed using an analytical balance (CP225D, Sartorius, Germany) with an accuracy of 10⁻⁴ g. Circulating oil solution from a thermostat (Huber CC1-505wl vpc55, Germany) used with an uncertainty of u(T) = 0.01 K controlled the temperature of the mixture. ¹H and ¹³C spectra were recorded on a 400 MHz (Bruker AVANCE 400) or 600 MHz (Bruker AVANCE 600) nuclear magnetic resonance spectrometer.
Figure 4.

Schematic of the HEL CrystalSCAN system for solubility measurement.

The content of Fer or Fer-Me in the microparticulate powders was determined by the following method [28 (link)]. The microparticles (about 5 mg) were accurately weighed using a high precision analytical balance (d = 0.01 mg; Sartorius, model CP 225D, Goettingen, Germany), and dissolved in methanol at 80 °C for 15 min. The samples were then cooled at room temperature, and the final volume of the solution was adjusted at 2 mL. Then, 10 μL of filtered solutions (regenerate cellulose, 0.45 μm) was injected into the HPLC system for Fer or Fer-Me quantification. The drug loading and entrapment efficiency were calculated according to the following Equations (2) and (3): $$\mathrm{Drug} \mathrm{loading} \left(\frac{\mathrm{W}}{\mathrm{W}}\right)(\%)=\frac{\mathrm{mass} \mathrm{of} \mathrm{drug} \mathrm{in} \mathrm{microparticles}}{\mathrm{mass} \mathrm{of} \mathrm{loaded} \mathrm{microparticles}}\times 100$$
$$\mathrm{Entrapment} \mathrm{efficiency} (\%)=\frac{\mathrm{mass} \mathrm{of} \mathrm{drug} \mathrm{in} \mathrm{microparticles}}{\mathrm{starting} \mathrm{mass} \mathrm{of} \mathrm{drug}}\times 100$$
All the values obtained are the mean of four independent experiments.

An accurately weighed amount of Fer or Fer-Me (about 0.6 mg weighed with the analytical balance Sartorius CP 225D), or microparticles containing an equivalent quantity of encapsulated substances, were added to 30 mL of PBS (pH 7.4). The samples were maintained at 37 °C and stirred mechanically (100 rpm). Aliquots (200 μL) were withdrawn at fixed time intervals, and 10 μL of filtered samples (regenerate cellulose, 0.45 μm) was injected into the HPLC system. An equal volume of medium was added after each sampling to maintain sink conditions. The collected samples were quantified for Fer and Fer-Me using the developed HPLC method. All the values obtained were the mean of four independent experiments.

Weight measurements were performed in a Sartorious CP225 D micro analytical balance (Göttingen, Germany). At each irradiation time, the mean value of three independent weight measurements was calculated and mass loss determined by subtracting the corresponding value obtained at t = 0 h. Final mass loss corresponds to the mean of the mass loss values of the three replicates for each typology (includes nine readings) converted to percentage of total initial weight. Samples were kept in a desiccator after irradiation until weight measurements were performed.

Body mass of the species collected during field sampling were measured to the nearest 0.01 mg using an electronic balance (CP225D; Sartorius Co, Arvada, CO, USA). Six individuals of each species were measured. The movement rate of individuals was measured by recording the time in took to move across cage enclosure tiles from edge to edge, Movement rate were recorded on the first day of experiments (N = 6 for each species).
To estimate grazer growth rates, the initial and dry body weights of larvae were measured before and after the experiment. Twelve larvae from each species were collected before the experiment began and dried at 55 °C for 24 h. The larvae were then weighed, and the dry weight per larva was calculated for each species. At the end of the experiment, all larvae were collected from cage enclosures, and preserved in 4% formaldehyde. These larvae were then dried and weighed as described above. The daily relative growth rates for each species (RGR) was calculated as follows:
${\displaystyle \mathrm{RGR}=\left[\left(\mathrm{final dry weight}\right)-\left(\mathrm{initial dry weight}\right)\right]∕12\left(\mathrm{d}\right).}$

For the purpose of estimating the amount of spinetoram residue, an Alliance 2695 Separations Module (Waters, Milford, MA, USA) was used in conjunction with a Micromass Quattro Micro triple-quadrupole mass spectrometer (Micromass, Manchester, UK). MassLynx software V4.1 and QuanLynx were used to control the instruments and analyse the data, respectively. Using a Turbo Vap LV device from Caliper Life Science, the samples were evaporated (Hopkinton, MA, USA). A high-speed chilled centrifuge, Model Avanti J-30I, was used to centrifuge the extracts (Beckman coulter, Brea, CA, USA). The rotor heads could contain eighteen 10 mL (JA-21) and eight 50 mL (JA-30.50 T1) samples. The samples and powder reagents were weighed using a top-loading balance with a digital display (Sartorius, CP 225D, Göttingen, Germany). Fluorinated ethylene propylene (FEP) centrifuge tubes (Nalgene, Rochester, NY, USA) of 10 mL and 50 mL were used, respectively, for the extraction and the dSPE clean-up processes. The final extracts were stored in standard 1.8 mL dark glass autosampler vials. For sample preparation, a homogenizer (Polytron, PT-MR-3100, Kinemetica AG, Malters, Switzerland) as well as a pH metre (CL 46 type, Toshniwal Instruments Pvt. Ltd., Chennai, India) were also employed.