Analysis of the free amino acids in the meat was carried out by the method of Mikami et al. (1994) . Each sample (5 g) was homogenized with distilled water (90 mL) for 1 min. After centrifugation for 20 min at 10,000g at 0℃, the supernatant was filtered through Whatman No. 5 and the samples were diluted 1:1 with 4 % trichloroacetic acid. The diluted samples were incubated at 37℃ for 30 min, and then filtered again with Whatman No. 5, repeatedly. The filtrate was then subjected to a final filtration with a 0.45 um Millipore filter. Each of the filtered samples was analyzed using a fully automated amino acid analyzer, HIT-ACHI L-8800A (Hitachi Ltd., Japan).
Whatman no 5
Manufactured by Cytiva
Sourced in United Kingdom
Whatman No. 5 is a type of qualitative filter paper. It is designed for general filtration purposes in the laboratory.
Automatically generated - may contain errors
Lab products found in correlation
1.2 1 3 300 mesh grids, by Quantifoil (1 mentions)
Graphene oxide solution, by Merck Group (1 mentions)
Mueller hinton agar (mha), by Cytiva (1 mentions)
Mueller hinton agar (mha), by Merck Group (1 mentions)
0.45 m filter unit, by Merck Group (1 mentions)
Waters 2695 hplc, by Waters Corporation (1 mentions)
Waters 2475, by Waters Corporation (1 mentions)
9 protocols using whatman no 5
1
Amino Acid Analysis of Meat
2
Graphene-Oxide Assisted Cryo-EM Sample Preparation
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Cryo-EM samples were prepared using graphene-oxide-coated Quantifoil 1.2/1.3 300 mesh grids (GO-grids). We produce our GO-grids as follows: we took an aliquot of the 2 mg/mL stock graphene-oxide (GO) solution (Sigma-Aldrich) and sonicated it for 3 min in a bath sonicator. This step is necessary to separate aggregated GO plates. After this, we prepared dilutions between 0.05 and 0.15 mg/mL for screening. Quantifoil grids were freshly glow-discharged and 4 µL of our GO dilutions were immediately applied to the grids. After 3 min incubation, the grids were blotted from the back side using Whatman No. 5 and washed twice with 15 µL water. The quality of the resulting GO-grids was immediately assessed in our Tecnai Spirit microscope. Typically, we looked for a compromise between grid coverage and number of GO layers, with most selected grids showing a few layers—sometimes single ones—and almost 100% grid coverage. Shortly before applying the sample, the GO-grids were mildly glow discharged for 10 s using a 5 mA current.
Vitrification was performed in Vitrobot Mark IV. Four µL of sample were placed onto a GO-grid, incubated for 1 min at 13 °C and 100% humidity, blotted for 3.5 s with a force of –3, and subsequently plunged into liquid ethane.
Vitrification was performed in Vitrobot Mark IV. Four µL of sample were placed onto a GO-grid, incubated for 1 min at 13 °C and 100% humidity, blotted for 3.5 s with a force of –3, and subsequently plunged into liquid ethane.
3
Antimicrobial Evaluation of Volatile Oils
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The antimicrobial activity testing was modified from Prabuseenivasan et al. [16 (link)]. Briefly, bacterial suspension was adjusted to McFarland standard No. 0.5 (1 × 108 CFU/mL) and spread over the Mueller-Hinton agar (MHA) plates using a sterile cotton swab. Each volatile oil was dissolved in 10% aqueous dimethyl sulfoxide (DMSO) with 0.5% v/v Tween 80 and sterilized by filtration. Sterilized disks (Whatman No. 5, 6 mm diameter) were impregnated with 20 μL of volatile oils and placed on the surface of MHA. The volatile dissolving buffer (10% aqueous DMSO, 0.5% v/v Tween 80) and tea tree oil were used as negative and positive control, respectively. After incubation at 37°C for 16-18 h, the inhibition zone was measured. All experiments were performed independently in triplicate and mean value was calculated.
4
Antibacterial Activity Assessment of Natural Compounds
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The antibacterial activity testing procedure was modified from Prabuseenivasan et al. [32 (link)]. Briefly, the bacterial suspension was adjusted to McFarland standard 0.5 (approximately 106 CFU/mL) and spread over Muller Hinton Agar (MHA; Merck, Darmstadt, Germany) plates using a sterile cotton swab. Each antibacterial material (R. officinalis L., C. sinensis L. and citric acid) was prepared at a 0.5% (w/v) concentration and sterilized by filtration. Sterilized disks (Whatman No. 5, 6 mm diameter) were impregnated with 20 µL of antibacterial materials and placed on the MHA surface; 10% DMSO used as a negative control. After incubation at 37 °C or 30 °C for 20 h, the inhibition zone was measured. All experiments were performed independently in triplicate, and the mean value was calculated.
5
Ethanol Extraction of Whole Dried CZ Plants
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One hundred gram of whole dried plants of CZ (Bioland, Cheonan, Korea) were extracted with 1,000 ml of 95% ethanol for 120 minutes under reflux and then filtered with filtering paper (Whatman No. 5; Whatman International Ltd., Kent, England, UK). The liquid from the filtering process was concentrated in a vacuum evaporator. The resultant residue was weighed as 19.5 g, and then dissolved in 70% ethanol for treatment for in vivo study subsequently. CZ extracts were then filtered to sterilize for in vitro study using 0.45 µm-filter units (Millipore, Burlington, MA, USA).
6
Antibacterial Activity Assay of Essential Oils
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Antibacterial activity was measured using the disc-diffusion method (Vanden Berghe and Vlietinck, 1991). The microorganisms were grown overnight at 37°C in 10 mL of Müeller Hinton broth (Bioxon 260-1, Estado de Mexico, Mexico). The cultures were adjusted to a turbidity comparable to McFarland no. 0.5 standard with sterile saline solution. Petri dishes containing Müeller Hinton agar (Bioxon, Edo. de Mexico, Mexico) were impregnated with these microbial suspensions. 5-mm diameter discs (Whatman no. 5) were saturated with 5 μL of essential oil. Discs of chloramphenicol (25 μg) were used as positive controls. The plates were incubated overnight at 37°C, and the diameters of any resulting inhibition zones (mm) were measured. Each experiment was repeated at least three times. Estimation of the minimal inhibitory concentration (MIC) was carried out using the broth dilution method (Vanden Berghe and Vlietinck, 1991). Dilutions of essential oil from 4000 to 62.5 μg/mL were used. Tubes were inoculated with a microorganism suspension of 105 CFU/mL. MIC values were defined as the lowest essential oil concentration that prevents visible bacterial growth after 24 h of incubation at 36°C. Chloramphenicol was used a reference. Each experiment was repeated at last three times.
The bactericidal kinetic assay was performed using appropriate concentrations of essential oil (corresponding to
The bactericidal kinetic assay was performed using appropriate concentrations of essential oil (corresponding to
7
Mycotoxin Extraction and Detection Protocol
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The samples (5 ml of liquid culture) were filtered through Whatman No. 5 (Whatman, Piscataway, NJ) filter paper and dried under nitrogen stream. The residues were dissolved into methanol water (3:1, v/v), adjusted to the pH value of 5·8–6·5 by 0·1 mol l−1 KOH water solution and cleaned using a SAX cartridge. The cartridge was conditioned with 5 ml of methanol followed by 5 ml of methanol–water (3:1, v/v). FB1 was eluted from the column to a glass collection vial with 10 ml of 1% acetic acid in methanol. The eluate was evaporated to dryness at 40°C under a stream of nitrogen. The cleaned sample was derivatized with OPA reagent (20 mg 0·5 ml−1 methanol diluted with 2·5 ml 0·1 mol l−1 disodium tetraborate (Na2B4O7 × 10H2O), then combined with 25 μl 2-mercaptoethanol) immediately before HPLC analysis by mixing the OPA reagent and the sample in ratio 4:1 v/v. After 2 min, the reaction mixture (10 μl) was injected in a HPLC C18 Nova Pak column (3·9 × 150 mm). Methanol–sodium dihydrogen phosphate (0·1 mol l−1 in water) solution (77:23; v/v) was adjusted to pH 3·35 with o-phosphoric acid and used as the mobile phase with the flow rate of 0·6 ml min−1. A Waters 2695 HPLC with a fluorescence detector (λEx = 335 nm and λEm = 440 nm, Waters 2475; Waters) was used for analysis.
8
Liquid Phase Release Evaluation of Oleogels
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The liquid phase release of oleogels made with CW and stored at room temperature (approximately 25 • C) for 10 days was evaluated using a technique similar to the one described by Dibildox-Alvarado et al. (2004) . The oleogels were placed on previously weighed filter paper (Whatman no. 5, 125 mm in diameter), and once the analysis time had elapsed, the gel was carefully removed, and the paper was weighed again. For each sample, four repetitions were carried out. The percentage of liquid phase release was calculated according to Equation (1), with the liquid phase release (LL), filter paper weight (fpw), weight of filter paper plus oleogel (fpw + g) and weight of the oleogel after storage (gwf ):
(1)
(1)
9
Fabrication of CNC-Reinforced Polymer Composites
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CNCs were isolated from the powdered cellulose (Whatman, No. 5) by acid hydrolysis with 65 wt% sulfuric acid, with mechanical stirring at 70 °C for 15 min [7] . After a dilution and centrifugation process, the crude CNC dispersion was dialyzed in distilled water and then in 7 wt% PEG aqueous solution. Ultimately, a refined CNC/water suspension was prepared at 23 wt% via finishing treatment using a homogenizer. The particle dimensions of CNC were ~110 nm in length and ~8 nm in diameter on average, when estimated by transmission electron microscopy. Surface sulfur content of the CNC particles approximated 0.65 wt%, when determined by an alkali titration method [7] .
The preparation of CNC suspensions in aqueous monomer (HEMA/water or HEMA/AA/water) was done by reference to a procedure described in an earlier paper [23] . Briefly, a weighed amount of the concentrated CNC/water suspension mentioned above was mixed with HEMA or HEMA/AA (95:5-70:30 in mol) and distilled water (diluent) in a light-blocked glass vial, using ultrasonic-wave treatment for 2 min. The weight ratio of monomer/water was adjusted to 1:1, and HMPPh and EDM were fed in the solvent, each at 0.5 wt%. 7-11 wt% CNC/monomer/water suspensions were thus prepared and stored at room temperature (25 °C) usually in a dark place.
The preparation of CNC suspensions in aqueous monomer (HEMA/water or HEMA/AA/water) was done by reference to a procedure described in an earlier paper [23] . Briefly, a weighed amount of the concentrated CNC/water suspension mentioned above was mixed with HEMA or HEMA/AA (95:5-70:30 in mol) and distilled water (diluent) in a light-blocked glass vial, using ultrasonic-wave treatment for 2 min. The weight ratio of monomer/water was adjusted to 1:1, and HMPPh and EDM were fed in the solvent, each at 0.5 wt%. 7-11 wt% CNC/monomer/water suspensions were thus prepared and stored at room temperature (25 °C) usually in a dark place.
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