In vitro drug release experiment was performed for SPR-BS and free SPR suspension (control) using the bag diffusion technique [29 (link)]. Dialysis bags (Visking®, MWCO 12,000–14,000, Serva, Denver, CO, USA) containing F3, F4, and SPR were immersed in PBS (pH 6.8) ensuring sink conditions [87 (link)]. Bags were placed in a shaking water bath (100 rpm) (Memmert GmbH, Schwabach, Germany) at 32 ± 0.5 °C [29 (link)]. At predetermined time intervals, samples were withdrawn and compensated with fresh medium. The amount of SPR released was determined using UV spectroscopy (Shimadzu, model UV-1601 PC, Kyoto, Japan) at 620 nm [85 (link)]. The experiment was conducted in triplicates and data were expressed as the mean ± SD. The SPR release mechanism from the developed nanocarrier was investigated using DD Solver 1.0 Software (Microsoft Excel add in program, Microsoft Corporation, Redmond, WA, USA) [88 (link)].
Visking mwco 12 000 14 000
Manufactured by Serva Electrophoresis
Sourced in United States, Germany
Visking®, MWCO 12,000–14,000 is a dialysis tubing made of regenerated cellulose. It is designed for molecular weight cut-off (MWCO) between 12,000 and 14,000 Daltons.
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5 protocols using visking mwco 12 000 14 000
1
In Vitro Drug Release of Lipid-Based Nanocarriers
2
Liposomal Entrapment Efficiency of Atazanavir
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The entrapment efficiency (EE) was indirectly estimated by measuring the concentration of free (unentrapped) ATV. Drug-loaded liposomal dispersion (1 mL) was placed in a dialysis bag (Visking®, MWCO 12,000–14,000; Serva, Germany) and dialyzed for 2 h against 70 mL of 5% ethanol in phosphate buffer (pH 6.8) at 2–8 °C [20 ]. ATV in the dialysate was then spectrophotometrically (Agilent Cary 60; Agilent Technologies, USA) determined at λmax of 242 nm [21 ]. Percentage EE and drug loading (DL; w/w) were then calculated using Eqs. 1 and 2 , respectively, where the whole weight was theoretically calculated from the actual weights of used ingredients.
3
In-vitro Release Profiles of Caffeinated Hyalurosomes
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In-vitro release profiles of the formulated caffeinated hyalurosomes and CAF either in solution form or loaded in a conventional gel were determined using the dialysis bag method (Abbas et al., 2022 (link); Simsolo et al., 2018 (link)). One ml sample of CAF aqueous solution, caffeinated hyalurosomes (F3 and F4 ) and CAF-GEL equivalent to 2% w/v CAF were placed in the dialysis bag (Visking®, MWCO 12000–14,000, Serva, USA), tied from both ends, immersed in 100 ml release medium (PBS, pH 5.5) (Abd et al., 2021 (link); Amasya et al., 2021 ; Iriventi and Gupta, 2020 ; Simsolo et al., 2018 (link)) and continuously shaken at 100 rpm in a thermostatic shaking water bath at 32 ± 0.5 °C (Memmert GmbH, Germany). At pre-determined time intervals (0, 1, 2, 3, 6, 8, 24 h), 2 ml samples were withdrawn from the release medium and compensated by an equal volume of fresh medium to maintain sink conditions. Samples were analyzed spectrophotometrically at λmax 273 nm (Arafa et al., 2020 (link)) against a buffer solution as blank. Each experiment was assessed in triplicates (Elsheikh et al., 2018 ; Gaafar et al., 2021 ).
4
In Vitro Dissolution of Lyophilized Formulations
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In vitro dissolution was performed for the selected lyophilized formulae (DNS-F7 and CS-DNS-F10) using an amount of drug equivalent to 5 mg in comparison to DCN powder. Dialysis bag diffusion technique was used.27 Dialysis membrane (Visking®, MWCO 12,000–14,000; SERVA, Heidelberg, Germany) was soaked in distilled water for 24 hours before the experiment. Samples were dispersed in 1 mL phosphate buffer in a dialysis bag of 5 cm length sealed hermetically and immersed in vials containing 15 mL sodium orthophosphate buffer 6.8 and were kept in a shaking water bath (Wisebath®, London, UK) at 37°C at 100 rpm. Samples were withdrawn at different time intervals up to 2 hours and replaced with an equal volume of dissolution medium. Samples were filtered and analyzed spectrophotometrically at the wavelength of 258 nm. Dissolution profile was expressed by the percentage diffused at 120 minutes (PD120), mean dissolution time (MDT), in addition to diffusion efficiency (DE).
5
In vitro Dissolution of Nano-Curcumin Formulation
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In vitro dissolution of NC-GL was performed on the selected lyophilized formulation; NC-GL7 compared to GL powder using the dialysis bag technique. Dialysis bags (Visking®, MWCO 12,000–14,000; SERVA, Heidelberg, Germany) were soaked in distilled water for 24 h prior to the experiment. Accurate amounts equivalent to 2 mg GL dispersed in 1 mL phosphate buffer pH 7.4 were filled in the dialysis bags which were then sealed and suspended in 50 mL phosphate buffer pH 7.4 containing CTAB 0.1% w/v to maintain sink conditions. The release medium was selected based on GL solubility study (data not shown). Samples were kept in a shaking water bath (Wisebath®, London, UK) at 37 °C at 100 rpm. At predetermined time intervals, 2.5 mL aliquots were withdrawn and replaced with fresh dissolution medium. Samples were analyzed spectrophotometrically (Cary 60 UV-Vis Spectrophotometer, Agilent, Santa Clara, CA, USA) at λmax 300 nm. Measurements were conducted in triplicate.
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