A stock inoculum suspension of Trichophyton rubrum was prepared using 14-days old cultures grown on Sabouraud’s dextrose agar (SDA) slants at 28 °C. The fungal colonies were covered with 2 mL of Sabouraud’s dextrose broth (SDB) followed by a gentle scratching of the agar’s surface with a sterile swab, resulting in a suspension of conidial and hyphal fragments that was transferred to a sterile tube and kept for 5 to 10 minutes at room temperature for sedimentation of heavy particles keeping micro-conidia suspended. Filtration was performed to retain hyphal fragments but permit the passage of dermatophyte micro-conidia to be used later (Motedayen et al., 2018 ). The turbidity of the fungal suspension was measured spectrophotometrically (ultraviolet/visible [UV/VIS] spectrophotometer; JASCO, Tokyo, Japan) at 600 nm and adjusted with SDB to match the 0.5 McFarland standard which is equivalent to 105-106 CFU/mL (Marcato et al., 2012 ).
Ultraviolet visible uv vis spectrophotometer
Manufactured by Jasco
Sourced in Japan
The Ultraviolet/Visible (UV/VIS) Spectrophotometer is a laboratory instrument used to measure the absorption or transmission of light in the ultraviolet and visible regions of the electromagnetic spectrum. It measures the amount of light absorbed or transmitted by a sample at specific wavelengths, providing information about the chemical composition and concentration of the sample.
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Lab products found in correlation
7 protocols using ultraviolet visible uv vis spectrophotometer
1
Preparation of Trichophyton rubrum Inoculum
2
In Vitro Release of Diosmin from PLGA Nanoparticles
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In vitro release of diosmin from the selected uncoated and chitosan-coated PLGA nanoparticles was studied in comparison with free diosmin using USP apparatus II (paddle method) (Dissolution Apparatus USP Standards, Scientific, DA-6D, Bombay, India). A dissolution medium consisting of 500 mL borate buffer pH 10.5 was kept at 37±0.5°C and stirred at 100 rpm. Accurately weighed sample of the lyophilized nanoparticles equivalent to 20 mg diosmin were introduced into the dissolution tester cells. At predetermined time intervals (0.5, 1, 2, 3, 4, 6, 8, 24, 48, and 72 h), 3 mL of the dissolution medium was withdrawn and replaced with an equal volume of a fresh dissolution medium. The collected aliquots were filtered and spectrophotometrically analyzed for drug concentration at 269 nm (ultraviolet/visible [UV/VIS] spectrophotometer; JASCO). The blank was the corresponding plain nanoparticles that were treated the same as the medicated. Each experiment was done in triplicate and the average percentage drug released was calculated to construct in vitro release curves.
3
In Vitro Dissolution of Optimized PLGA NPs
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The in vitro dissolution study of SPL from the optimized PLGA NPs compared to free SPL was conducted by following the protocol of the USP apparatus II (paddle method) (Dissolution Apparatus USP Standards, Scientific, DA-6D, Bombay, India).30 (link),35 (link) Accurately weighed samples of the freeze-dried SPL NPs equivalent to 10 mg drug were added to the vessels containing 500 mL of 0.1N HCl (pH 1.2) or 0.066M phosphate buffer (pH 6.8) with paddle rotation speed of 100 rpm at 37±0.5 °C. At different time intervals (0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 12, and 24 h), 3 mL of the dissolution medium were withdrawn and replaced with an equivalent volume of the fresh medium. The collected samples were filtered and analyzed spectrophotometrically for drug amount at 242 nm against the blank of plain NPs treated the same (ultraviolet/visible [UV/VIS] spectrophotometer; JASCO, Tokyo, Japan). The experiments were performed three times, and the average drug release % was calculated at each time interval and plotted against time to construct the in vitro dissolution curves.
4
Quantifying Nail Clipping Drug Absorption
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To quantify the absorbed and desorped CIX, the nail clippings of groups 2 and 3 were washed with deionized water then methanol to remove any drug traces on the surface. They were dissolved in 1 mL of 1 M sodium hydroxide by stirring (MS300HS, MTOPS Corp., Korea) overnight and then filtered through 0.22 µm syringe filter. The filtrate was properly diluted with methanol and CIX accumulated in the nail clippings was determined spectrophotometrically at 248 nm (ultraviolet/visible [UV/VIS] spectrophotometer; JASCO, Tokyo, Japan) against the respective blank treated the same. The enhancement factor (EFnail) indicating the improvement of CIX penetration into the nail clippings from HPCH lacquer compared to the drug solution was calculated according to the following equation (Palliyil et al., 2013 (link)):
5
Spectrophotometric Analysis of Naringin Encapsulation in PF68 Micelles
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Methanol was used to disrupt the micellar structure to be then analyzed spectrophotometrically (ultraviolet/visible [UV/VIS] spectrophotometer; JASCO, Tokyo, Japan) for drug content at 281 nm vs plain micelles treated the same. The equations used for the estimation of LE% and EE% of naringin in PF68 micelles were as follows:43 (link)
Further characterization of polymeric micelles with PF68 showing the highest EE% would be carried out.
Further characterization of polymeric micelles with PF68 showing the highest EE% would be carried out.
6
Spectrophotometric Analysis of Hesperetin Micelles
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Dimethylsulfoxide (DMSO) was used to disrupt the micellar structure to be then analyzed spectrophotometrically (ultraviolet-visible [UV-Vis] spectrophotometer; JASCO, Tokyo, Japan) at 231 nm for drug content. The equations we used for the estimation of LE and EE% of Hesperetin in P5DF10, P10DF10, P20DF10 micelles were as follows (Zu et al., 2011 (link); Mohamed et al., 2018 (link)):
7
In Vitro Dissolution Study of Coated and Uncoated Powders
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In vitro dissolution study was carried out by USP apparatus II paddle method (Dissolution Apparatus USP Standards, Scientific, DA-6D, Bombay, India). The experiment was performed in different pH values of GIT, 0.1N HCl (pH 1.2), phosphate buffer (pH 6.8) and phosphate buffer (pH 7.4). Eight mg of CC and the equivalent from uncoated F7-SMCC and coated C1-SMCC powders, were subjected to the in vitro dissolution test. Each powder was transferred to 500 mL of different dissolution media,14 (link) each containing 0.35% Tween 20 (to achieve sink condition) and stirred at 100 rpm at 37°C ±0.5. Aliquots of 5 mL were withdrawn at predetermined time intervals (0.5, 1, 2, 3, 4, 5, 6, 7, and 8 h), and replaced by same volume of fresh media. The solutions were filtered through 0.45 µm Millipore filter and the concentration of CC was measured by UV spectrophotometry spectroscopy (ultraviolet/visible [UV/VIS] spectrophotometer, JASCO, Tokyo, Japan) at predetermined λmax 254, 257, and 255 for pH values 1.2, 6.8 and 7.4,14 (link) respectively using blank of plain formulations treated similarly. Each experiment was done in triplicate.
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