The thermal activation process was carried out in a custom-made 30 mm diameter quartz tube attached to a sample flask, and the whole setup was connected to a vacuum system. In a typical experiment, 100 mg MWNTs were heated to 1000°C at a constant ramping rate of 10 °C/min under vacuum (~ 5 × 10−4mbar), in a three-zone tube furnace (PTF 12/38/500, Lenton Ltd, UK) and held at the activation temperature for 2 hours. After the activation step, the quartz tube was slowly removed from the heating zone and allowed to cool to room temperature under vacuum. The MWNTs were then transferred to the connected round bottom flask by gravity. 8 mL of the reactant was then injected into the flask containing the thermally-activated MWNTs. The reaction mixture was stirred at room temperature overnight. The unreacted reactant was removed via filtration through a 0.45 µm pore size polytetrafluoroethylene (PTFE) membrane (Whatman, UK) under vacuum. The product was thoroughly washed with 3 × 90 mL of washing solvent, then dispersed in 90 mL of solvent and bath sonicated (USC300T, 45kHz, 80W, VWR International, USA) for 15 minutes. The filtration-sonication cycle was repeated three times in order to remove any physically absorbed reactants. The functionalised MWNTs are named by the abbreviation of the grafted polymer: e.g. P(MMA)-MWNT. The other sample codes can be found in Table 1 .
Ptfe membrane
Manufactured by Cytiva
Sourced in United Kingdom
The PTFE membrane is a porous, hydrophobic membrane made of polytetrafluoroethylene (PTFE). It is designed for use in various laboratory and industrial applications that require filtration, separation, or gas/liquid exchange.
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6 protocols using ptfe membrane
1
Thermal Activation and Polymer Grafting of MWNTs
2
Ultrasound-Assisted Extraction of A. tinctoria Roots
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For the three experiments, primary and secondary roots of A. tinctoria plants were freeze-dried separately and reduced into powder using liquid nitrogen, mortar and pestle. Fifty milligram of each root type was then mixed in glass test tube (labbox 10 ml neutral glass, TU04-160-100) closed with a plastic cap. Then, the total 100 mg of root material was subjected to a 20-min ultrasound-assisted extraction (two cycles), using n-hexane 97% (2 ml) at room temperature. The samples were finally centrifuged at 1,500 rpm for 10 min at 4°C and the supernatants of each cycle were combined and evaporated at room temperature. The extraction protocol was adapted from Bossard et al. (2022) (link).
Prior to analysis, the plant extracts and nutrient solution residues were weighed and solubilized in methanol and were filtered through a 45-μm PTFE membrane (Whatman™, Maidstone, UK). Each sample was adjusted to the final concentration of 2 mg ml–1, using methanol LCMS-grade.
Prior to analysis, the plant extracts and nutrient solution residues were weighed and solubilized in methanol and were filtered through a 45-μm PTFE membrane (Whatman™, Maidstone, UK). Each sample was adjusted to the final concentration of 2 mg ml–1, using methanol LCMS-grade.
3
Enrichment of (6,5) SWCNTs via Polymer Wrapping
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Shear-force
mixing (Silverson L2/Air, 10,230 rpm) of 50 mg CoMoCat raw material
(Sigma-Aldrich, MKCJ7287) in a solution of 65 mg of PFO-BPy (poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-(6,6′-(2,2′-bipyridine))], American Dye
Source, Inc., Mw = 40 kg mol–1) in 140 mL of toluene for 72 h at 20 °C was used to prepare
nearly monochiral (6,5) SWCNT dispersions, as described previously.6 (link) Undispersed amorphous carbon and other SWCNT
chiralities were separated from the PFO-BPy-wrapped (6,5) SWCNTs by
centrifugation (twice, 45 min at 60,000g, Beckman
Coulter Avanti J26XP centrifuge) and filtration through a syringe
filter (Whatman PTFE membrane, pore size 5 μm). To remove excess
PFO-BPy, the polymer-rich stock dispersion was vacuum-filtered through
a polytetrafluoroethylene membrane (Merck Millipore, JVWP, pore size
0.1 μm). The obtained filter cake was washed three times in
10 mL of toluene at 80 °C for 10 min and redispersed in fresh
toluene by bath sonication for 30 min. Absorption (Cary 6000i UV–vis–NIR
spectrometer, Varian Inc.) and Raman spectroscopy (InVia Reflex, Renishaw
plc) confirmed the chiral purity and low content of the unbound polymer
of the (6,5) SWCNT dispersion (see Figure S1,Supporting Information ).
mixing (Silverson L2/Air, 10,230 rpm) of 50 mg CoMoCat raw material
(Sigma-Aldrich, MKCJ7287) in a solution of 65 mg of PFO-BPy (poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-(6,6′-(2,2′-bipyridine))], American Dye
Source, Inc., Mw = 40 kg mol–1) in 140 mL of toluene for 72 h at 20 °C was used to prepare
nearly monochiral (6,5) SWCNT dispersions, as described previously.6 (link) Undispersed amorphous carbon and other SWCNT
chiralities were separated from the PFO-BPy-wrapped (6,5) SWCNTs by
centrifugation (twice, 45 min at 60,000g, Beckman
Coulter Avanti J26XP centrifuge) and filtration through a syringe
filter (Whatman PTFE membrane, pore size 5 μm). To remove excess
PFO-BPy, the polymer-rich stock dispersion was vacuum-filtered through
a polytetrafluoroethylene membrane (Merck Millipore, JVWP, pore size
0.1 μm). The obtained filter cake was washed three times in
10 mL of toluene at 80 °C for 10 min and redispersed in fresh
toluene by bath sonication for 30 min. Absorption (Cary 6000i UV–vis–NIR
spectrometer, Varian Inc.) and Raman spectroscopy (InVia Reflex, Renishaw
plc) confirmed the chiral purity and low content of the unbound polymer
of the (6,5) SWCNT dispersion (see Figure S1,
4
Bacterial Capture and Growth Monitoring
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As illustrated in Figure 1 a,b a values, the signal intensities of the maxima (I±m) are obtained. A pinhole (diameter = 3 mm) in the collimated single mode laser beam (Thorlabs LPS-635-FC with collimator CFC2-A, λ = 635 nm, waist diameter = 360 µm (1/e2 width)) is directed onto the grating by a parabolic mirror, which leads to the generation of a diffraction pattern on a sCMOS sensor (panda, PCO AG, Kelheim, Germany). By local integration of the gray mirror that directs the diffracted light to the camera suppresses the intense central maximum. The fluidic flow and the bacterial loading of the chip were realized by eight pneumatic controllers (MFCS-EZ, Fluigent, Le Kremlin-Bicêtre, France), which were connected by polytetrafluoroethylene (PTFE) tubes (ID = Ø 180 µm, Techlab, Germany) to eight sample vials (D1.1 to R8) (Biozym Scientific GmbH, Hessisch Oldendorf, Germany). A 5 µm PTFE membrane (Whatman, Maidstone, UK) between the tube outlets and the chip inlets acted as a filter to prevent clogging. To switch between bacterial capturing and growth mode, a three-way valve (2-Switch, Fluigent, Le Kremlin-Bicêtre, France) was used, as shown schematically in Figure 2 a. The fluidic setup is integrated into the optical setup and is surrounded by an opaque housing, which was heated to 37 °C during the experiments by an incubator (ES-20, Grant Instruments Ltd., Cambridge, UK).
5
Multisolvents Extraction of Snake Venom
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The extraction step consists of sonication-assisted four-solvent process (in order: n-heptane, dichloromethane, ethyl acetate, methanol). The venom was grounded up finely and transferred into a borosilicate glass conical-bottom recipient and the first solvent (i.e., n-heptane) was added (ratio 1:20 m/v). The recipient was briefly vortexed and then placed in an ultrasonic bath (Branson® 3510, Danbury, CT, USA) for 60 min at room temperature. After being allowed to settle for 15 min approximately, the supernatant was filtered on a PTFE membrane ( = 0.45 μm, Whatman, Buckinghamshire, UK) and stored in a tared brown-glass vial. The venom pellet was placed under a gentle stream of nitrogen (Sample Concentrator EVAEC1-S, VLM, Bielefeld, Germany) until completely dry and the process was repeated with the following solvents. The four extracts were evaporated to dryness under nitrogen. Extraction yields (%, m/m) were evaluated by calculating the following ratio: (dry residue mass/ground-up venom mass)*100.
6
Preparative TLC Fractionation of Plant Extract
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Fraction F2 that was obtained following the fractionation of the methanolic crude extract underwent a supplementary fractionation step. For this purpose, preparative thin-layer chromatography was performed. An aliquot of the liquid fraction sample was deposited manually on the silica plate (PLC silica gel 60 F254 2 mm, Merck, Darmstadt, Germany). The mobile phase was composed of the upper phase of a n-butanol/water/ethyl acetate (20/10/5) mixture. After visualization under UV at 254 nm, the zones of interest were scratched. The scraped silica was extracted with methanol by vortexing and ultrasonication for 60 min. After centrifugation, the supernatant was filtered on a PTFE membrane ( = 0.45 μm, Whatman, Buckinghamshire, UK) and stored in a tared brown-glass vial. Each sub-fractions were evaporated to dryness under nitrogen.
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