The antibacterial efficacy of the set specimens against Streptococcus mutans (ATCC 25,175) was investigated in vitro using standard disc diffusion method on brain heart infusion (BHI) agar plates. Cement discs (6 mm diameter, 3 mm thickness) were prepared with sterile instruments and sterilized by UV radiation (CAMAG UV Cabinet, CAMAG Germany) at 254 nm for 60 min before proceeding into the next step [29 (link)]. Mueller–Hinton agar plates (Sigma Aldrich, MO, USA) seeded with 1.8 × 108 cfu/mL (0.5 OD600) of the test bacteria were checked for the presence of inhibition zones after 24 h of incubation at 37 °C. Inhibition zones surrounding the specimens were measured (mm) using a digital caliper at the outer limit of the inhibition zone generated considering only halos > 6 mm.
Uv cabinet
Manufactured by CAMAG
Sourced in Germany, Switzerland, United States
The UV cabinet is a laboratory equipment used for visualizing and documenting fluorescent compounds. It provides a controlled environment with UV light sources to observe and analyze samples. The cabinet allows for the examination and documentation of samples under specific UV wavelengths.
Automatically generated - may contain errors
Lab products found in correlation
Avance dmx400 ft nmr spectrometer, by Bruker (2 mentions)
Acetone, by HiMedia (2 mentions)
N hexane, by HiMedia (2 mentions)
Tlc jar, by CAMAG (2 mentions)
Automatic tlc coater, by CAMAG (2 mentions)
Mueller hinton agar plate, by Merck Group (1 mentions)
8 w uv lamp, by CAMAG (1 mentions)
Eos d60, by Canon (1 mentions)
R 200, by Büchi (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Silica gel f254, by Merck Group (1 mentions)
Waters acquity uhplc system, by Waters Corporation (1 mentions)
Twin trough chamber, by CAMAG (1 mentions)
Linomat 5 applicator, by CAMAG (1 mentions)
Tlc scanner 4, by CAMAG (1 mentions)
Tlc glass plates, by CAMAG (1 mentions)
Compound microscope, by Olympus (1 mentions)
Silica gel type g, by Merck Group (1 mentions)
13 protocols using uv cabinet
1
Antibacterial Efficacy of Dental Materials
2
Visualizing Polymer Layer Thickness
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For UV measurements an 8 W UV-lamp (Camag; Berlin, Germany) was used at 254 nm wavelength with a camera EOS D60 (Canon; Krefeld, Germany) for imaging in manual mode with the settings f-number F5.6, ISO1000, focal lengths 18 mm and shutter speed 1 s, carried out in a Camag UV-cabinet (Camag; Berlin, Germany). We increased the contrast between the PAN-support membrane and the PS-b-P4VP top layer by using a UV-lamp (254 nm). While the PAN backscatters parts of the UV-light, which then appears purple, PS-b-P4VP partially absorbs this wavelength and appears black. If the PS-b-P4VP layer thickness is low the UV-light will penetrate this layer, backscatter at the PAN-support membrane, and not absorb completely on its way back. In consequence, deviations of PS-b-P4VP layer thickness can be seen as purple light with changing intensity. To gain this effect, the shutter speed of the camera was increased to 1 s, which was enough for our purpose.
3
Qualitative Analysis of Botanical Extracts
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TLC using the method with external standards [10] was used for the qualitative analysis of chemical constituents in all extracts. Nine extracts, four polar standards (ascorbic acid, ellagic acid, gallic acid, and quercetin), and nonpolar standards (lauric acid, monolaurin, dilaurin, and trilaurin) were separately dissolved in an appropriate solvent to obtain a solution having a concentration of about 1 mg/ml. Each 5 µl of extract and standard solution was individually spotted onto silica gel 60 F254 TLC plates (250 µm thickness). The plates were separately developed in dichloromethane:methanol (7:3, v/v) and hexane:dichloromethane:ethyl acetate (8:1:1, v/v/v) for the detection of polar and nonpolar substances respectively. The mobile phases were modified from our previous method [11] . After drying, the TLC plates were examined under UV light at 366 and 254 nm (Camag UV cabinet, USA), exposed to iodine vapor, and then sprayed with a freshly prepared solution of p-anisaldehyde in sulfuric acid. The Rf values of the interesting spots were calculated.
4
Chromatographic and Spectroscopic Analysis of Organic Compounds
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Organic solvents were removed using a rotary evaporator R-200 (Buchi, Switzerland). UV cabinet (CAMAG, Muttenz, Switzerland) was used to view the TLC chromatograms. 1H-NMR and 13C-NMR spectra were recorded at room temperature on a Bruker Avance DMX400 FT-NMR spectrometer (Bruker, Billerica, MA, USA), operating at 400 MHz for 1H and 100 MHz for 13C at room temperature using deuterated MeOD-d4, with tetramethylsilane (TMS) as the internal standard. Spin multiplicities were reported as following: s (singlet), d (doublet), dd (doublet of doublets), and dt (doublet of triplets). ESI-MS were recorded on an Ultimate 3000LC-MS. The measurement was carried out by an electrospray ionization method in a positive mode with the source voltage and temperature being fixed at 3 kV and 250°C.
5
Thin Layer Chromatography Analysis of Extracts
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TLC analysis of extracts was with a stationary phase of TLC silica gel 60 F254 aluminium sheets, a migratory distance of 8 cm. Five microliters of each extract solution (20 mg/mL) was applied on a TLC plate. Mobile phase systems were chloroform: methanol: water: glacial acetic acid (6:5:1:1, v/v/v/v) and chloroform: methanol: glacial acetic acid (7:3:1, v/v/v). The TLC chromatograms were detected under visible light and short-wave UV light at 254 nm (Camag UV cabinet, USA). The spots on the plates were then exposed to different visualizing reagents such as Dragendorff's reagent, 10% v/v sulfuric acid - anisaldehyde, DPPH, and iodine vapor [16 ]. The detected spots were recorded according to their retardation factor (Rf) values.
6
HPTLC Analysis of Plant Flavonoids
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High-performance thin layer chromatography (HPTLC) analysis was performed using the instrument from CAMAG (Germany). Thin layer chromatography (TLC) plates (silica gel >60 F254, 20 cm × 10 cm; Merck) were prewashed with methanol. The plates were activated in an oven at 100°C for 10 min. Ten microliters of individual plant extracts (1 mg/ml) was spotted onto the precoated plates using Linomat 5 application system. Rutin hydrate (50, 100, 200 μg/ml) was used as the marker flavonoid. The flavonoids were separated using ethyl acetate: Formic acid: Glacial acetic acid: Water (100:11:11:27) as the mobile phase. Natural product (NP) reagent was used as the flavonoid derivatizing agent, and the spots developed were visualized under CAMAG UV cabinet (366 nm) and digitized using CAMAG photodocumentation system.
7
Quantification of Cellulose-Producing Mutants
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The number of Cel− mutants was determined in the culture medium taken from the bioreactor during the fermentation process by performing quantitative plating as describe in the section Determination of the number of living cells. Next, the grown colonies were flooded with 20 mL of PBS with 0.01% of Tinopal LPW dye (Calcofluor White M2R, Tinopal UNPA-GX, MilliporeSigma, Burlington, MA, USA) and incubated for 24 h in darkness. The colonies were examined using an UV Cabinet (CAMAG, Muttenz, Switzerland) at a wavelength of 366 nm. BC fibrils stained with a fluorescent dye were observed only around the cellulose-producing colonies (in contrast to the Cel− mutants).
8
Bioactive Metabolite Purification from Fungal Extract
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Partial purification of the bioactive metabolites of the CEL7 extract was performed using thin-layer chromatographic techniques. The fungal extract was prepared as described earlier. The dried fungal extract was resuspended in EA, maintaining a concentration of 20 mg ml−1, and 10 µl of EA extract was loaded onto alumina–silica Thin Layer Chromatographic (TLC) plates (MERCK Silica Gel F254) using capillary glass tubes. Acetone (HiMedia) and n-hexane (HiMedia) in a ratio of 2:8 were used as running solvents, and the retention factor was calculated for all the bands under UV light using a Camag UV Cabinet. The bioactive compounds corresponding to each band were scratched and collected, and they were finally dissolved in 1 ml of EA, which was centrifuged (7,000 rpm for 15 min) and evaporated to dryness. The dried components were then dissolved in DMSO at 100 µg ml−1 concentration, and the disk diffusion technique measured antifungal action (Wang et al., 2014 (link)).
9
Analytical Techniques for Chemical Characterization
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A Rotary evaporator R-200 (Buchi, Switzerland) was used to remove organic solvents. TLC chromatograms were viewed under a UV cabinet (CAMAG, Muttenz, Switzerland), 1H-NMR and 13C-NMR spectra were recorded at a room temperature on a Bruker Avance DMX400 FT-NMR spectrometer (Bruker, Billerica, MA, USA). MS was run on an ultra-high-performance liquid chromatography-mass spectrometer (UHPLC-MS) Waters® Acquity UHPLC system (Waters, Milford, MA, USA) equipped with electrospray ionization (ESI) in the negative mode.
10
Quantification of Tannins in Terminalia chebula
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Quantification of ellagic acid and gallic acid from the aqueous extract of T. chebula was performed by the High-Performance Thin Layer Chromatography (HPTLC) method. HPTLC analysis was carried out by using CAMAG HPTLC (Switzerland) equipped with visionCATS HPTLC Software, CAMAG Linomat V applicator, CAMAG Hamilton Micro syringe (100 µL), CAMAG TLC Scanner 4 (4.6 mm × 25 cm, 5 μm), CAMAG UV Cabinet (254 and 366 nm), and CAMAG Twin Trough Chamber (20 × 10 cm). The separation was carried out using a solvent system of Methanol: Water: Formic acid (4:6:0.5) v/v/v, and the stationary phase was pre-coated alumina backed, HPTLC Plate Silica gel 60 RP-18 (Merck). The scanning of the developed plate was performed at 254 nm. Ellagic acid and gallic acid reference material were used as standards for quantifying tannins in the extract. The standard solutions of ellagic acid and gallic acid of the concentration of 100 μg/mL were prepared in methanol and used for HPTLC analysis.
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