The process oil, toluene and water were selected as sonication media in this investigation. The prepared GRs were suspended in media and then exposed to sonication using a probe ultrasonic device (BANDELIN SONOPULS HD3100, HF power of maximum 100 W, microtip MS 73, Italy). The sonication power changed ranging from 30 W to 60 W and sonication time altered between 10 and 30 min. A control sample was prepared by mechanical devulcanization via two roll milling of the GR sample for 20 min (i.e. DGR-Ctrl). To understand the swelling effect on devulcanization yield, GRs were also pre-immersed in oil, toluene and water for 0, 24 and 48 h.
Hd3100
Manufactured by Bandelin
Sourced in Germany
The HD3100 is a laboratory equipment designed for high-frequency ultrasonic cleaning. It features a digital timer and temperature control for precise operation.
Automatically generated - may contain errors
9 protocols using hd3100
1
Ultrasonic Devulcanization of Rubber
2
Microbial Cell Lysis Protocol
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Using the reporter assays as indicators, a basic method for lysate production comprising only the most necessary processing steps was established (experiments 1 –3 , Fig. 1 ). This method was modified (Table A.1 ) to test the influence of single experimental parameters on enzymatic activities, which were assessed using the target analyte assay.
Ultrasonication (physical disruption) was applied as a standard method for the disruption of microbial cells from activated sludge. Cell lysis was performed with a sonication microtip (Bandelin MS73 connected to ultrasonic transducer HD3100) at 10 mm immersion depth in 50 mL tubes filled with 25 mL of concentrated sludge (∼19 gTSS L−1 in HN-buffer). Sonication was applied to the ice cooled samples with a power density of 1 W/mL for 10 min (net) in intervals of 15 s with 15 s breaks to avoid sample heating.
Cell debris was removed by centrifugation for 20 min at 14,000 rpm, 4 °C (Sigma 3K30, rotor 19776-H) and the supernatant, i.e. the crude cell lysate, was then filtered through 0.2 μm syringe filters (Whatman 25 mm GD/X polyethersulfone (PES) membrane with glass microfiber prefilter) to remove residual cells or cell debris. The resulting cell-free lysate (in HN-buffer) was kept on ice at all times and analyzed the same day.
Ultrasonication (physical disruption) was applied as a standard method for the disruption of microbial cells from activated sludge. Cell lysis was performed with a sonication microtip (Bandelin MS73 connected to ultrasonic transducer HD3100) at 10 mm immersion depth in 50 mL tubes filled with 25 mL of concentrated sludge (∼19 gTSS L−1 in HN-buffer). Sonication was applied to the ice cooled samples with a power density of 1 W/mL for 10 min (net) in intervals of 15 s with 15 s breaks to avoid sample heating.
Cell debris was removed by centrifugation for 20 min at 14,000 rpm, 4 °C (Sigma 3K30, rotor 19776-H) and the supernatant, i.e. the crude cell lysate, was then filtered through 0.2 μm syringe filters (Whatman 25 mm GD/X polyethersulfone (PES) membrane with glass microfiber prefilter) to remove residual cells or cell debris. The resulting cell-free lysate (in HN-buffer) was kept on ice at all times and analyzed the same day.
3
Sheep RBC Hemolysis Assay for Particle Toxicity
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RBCs were purified from sheep blood in Alsever’s solution by centrifugation at 3,000 × g for 2 min (Rotina 380R; Hettich, MA) and washing three times with .9% NaCl. RBCs were suspended in .01 M PBS at the final concentration of 5% by volume. Sheep RBCs were used because they showed a sensitivity to silica very similar to that of human RBCs (Arienzo and Bresciano, 1969 ). Particles were dispersed in .01 M PBS and sonicated 2 min on ice (horn, 3 mm; frequency, 20 kHz; maximum power output, 25 W; amplitude, 120 μm; Sonopuls HD 3,100, Bandelin, Germany), just before testing. Serial dilutions of the starting particle dispersions were performed according to the final surface area doses used for experiments. Dispersions were distributed in quadruplicate in a transparent 96-well plate (150 µL/well), and the RBC suspension was then added (75 mL/well). Negative and positive controls consisted of .01 M PBS and .1% Triton-X 100 in PBS, respectively. The plate was incubated on a plate shaker at 37°C for 30 min, and then centrifuged at 216 × g for 5 min. Supernatants were transferred to a new plate (75 mL/well), and the absorbance of the hemoglobin released was determined at 540 nm on a UV/vis spectrophotometer (Ensight, Perkin-Elmer, Waltham, MA) using the software Kaleido 2.0 (Perkin-Elmer).
4
Synthesis of Zinc Oxide Nanoparticles
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For this purpose, 3.65 g Zn (NO 3 ) 2 .6H 2 O (Loba chemie, India) was added to 100 mL of water. The solution was placed on a mixer (Heidolph model, Germany) at 450 rpm for 30 minutes at 25 °C and its pH was adjusted to 10 using NaOH. The prepared solution was then placed in an ultrasonic device (Bandelin model HD 3100) with water circulation within 2 hours. Thereafter, the prepared ZnO was separated by centrifuge and dried in an oven within 24 hours (24Pirhashemi and Habibi-Yangjeh 2017).
5
Comprehensive Materials Characterization
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The phase structure of the materials was studied by Philips Xpert XRD, applying CuKα radiation. The morphological features and chemical composition of the samples were characterized by LEO 1430 VP SEM/EDX instrument. The FT‐IR spectra were provided by a PerkinElmer Spectrum RX I instrument. The ultrasonic treatment was applied with a Bandelin ultrasound generator HD 3100.
6
Ultrasonic Synthesis of Pure Zincite
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Zinc Oxide (uncoated, pure synthetic zincite) 44 158* (SONOPULS HD 3100, Bandelin, Germany) for 10 min with 10% amplitude and energy of
7
Thyroglobulin Protein Quantification
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Culture condition, treatment, and decapsulation have been described above. Before h-thyroglobulin (Tg) measurement, cell lines were physically lyzed by sonication with 15-W power for 30 s with consecutive 1 s ON and 1 s OFF power (Bandelin, model HD3100). The amount of Tg protein was measured by IRMA Kit (Isotopes Co., Ltd. (IZOTOP), Budapest, Hungary), based on the manufactureʼs guideline.
8
Nanoreservoirs for Sustained Drug Delivery
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Nanoreservoirs were firstly prepared employing Compritol 888 ATO as a sustained release lipid matrix by melt–emulsification–ultrasonication method (Li et al. 2021 (link)). Briefly, 500 mg the lipid was blended with CA-ODA conjugate equivalent to 10 mg CA on a heater of 85 °C. Simultaneously, 300 mg Pluronic F-127 in 5 mL deionized water was heated to 85 °C as well. Afterwards, Pluronic F-127 solution was gently added to and homogeneously dispersed with the drug-lipid melt. The obtained coarse hot o/w emulsion was ultrasonicated at 60% amplitude for 15 min using probe sonicator (Bandelin HD 3100, Germany). Finally, CA-NRs were developed by allowing hot nanoemulsion to cool in an ice bath. After that, 500 µl of CHS solution of different concentrations (0.5–5 g%) was mixed with the selected CA-NRs and stirred at 25 °C for 1 h and then stored at 4 °C. Plain nanoparticles were prepared using the same procedure without addition of drug conjugate to the lipid phase or surface functionalization.
9
Stabilizing Naked Nanoparticles with Sodium Citrate
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The synthesized nude nanoparticles (i.e. without any capping agent) were not stable in aqueous suspension. To enhance their stability, a layer of sodium citrate was grafted onto the nanoparticle surface. 20 mL of sodium citrate solution (2 M, in water) were added to the nanoparticles (10 mL) and the suspension was heated at 80 °C under vigorous stirring for 1 hour on the heating plate. Soon after, the suspension was ultrasonicated (using an ultrasonic homogenizer Bandelin HD3100) for 30 min at 20 kHz, at the maximum power output. Immediately after that, the nanoparticles were washed by magnetic decantation after acetone addition (5 times) and finally resuspended in 5 mL of sodium citrate (5 mM water, as stabilizer buffer for storage). The suspension was then placed for 20 min in an ultrasound bath (56 kHz) at 60 °C to facilitate the evaporation of any trace of acetone. Finally, the sample was filtered by using a nylon syringe filter, with a pore size of 0.4 µm, to remove any possible large aggregate in the suspension.
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