Model v 530

Manufactured by Jasco

Sourced in Japan, Italy

The Model V-530 is a spectrophotometer designed for accurate absorbance measurements across a wide range of wavelengths. It features a high-resolution monochromator and a sensitive photodetector to provide precise and reliable data.

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Lab products found in correlation

Cellulose nitrate filter, by Sartorius (2 mentions) Cellulose acetate filter, by Sartorius (1 mentions) Prism software version 8, by GraphPad (1 mentions) Z 233 mk 2, by Hermle (1 mentions) Jms s3000 spiral tofms, by JEOL (1 mentions)

11 protocols using model v 530

Quantifying Pluronic F68 in H-NMeds

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The residual amount of surfactant in the H-NMeds was determined by a colorimetric method [39 (link)]. Briefly, ~1 mg of a freeze-dried H-NMeds sample was solubilized in 0.5 mL of dichloromethane. Then, 10 mL of distilled water was added and the organic solvent was evaporated at r.t. under stirring for 2 h. The suspension was filtered (cellulose acetate filter, porosity 0.45 µm, Sartorius, Florence, Italy) to obtain an aqueous solution (A).
To calculate the amount of Pluronic^® F68 in the formulation, 2 mL of the aqueous solution (A) was treated with 2 mL of 0.5% (w/v) BaCl₂ in HCl 1 N and 0.5 mL of an aqueous solution of I₂/KI (0.05 M/0.15 M). The obtained solution was incubated at r.t. for exactly 10 min in the dark. Pluronic^® F68 concentration was determined measuring the absorbance at 540 nm (Model V530, Jasco, Cremella, Italy). A calibration curve was calculated using the same method on stock solutions of Pluronic^® F68, and linearity was found in the range of 2–18 µg/mL. All data are expressed as the mean of at least three determinations. The amount of surfactant in the formulation was expressed as the Pluronic^® F68 Content % (PC %) and calculated using the following equation:

Ottonelli I., Duskey J.T., Rinaldi A., Grazioli M.V., Parmeggiani I., Vandelli M.A., Wang L.Z., Prud’homme R.K., Tosi G, & Ruozi B. (2021). Microfluidic Technology for the Production of Hybrid Nanomedicines. Pharmaceutics, 13(9), 1495.

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Quantifying Total Phenolic Content

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The total phenolic content of the methanolic extracts was determined using the Folin-Ciocalteu method, as previously described by Faustino et al. (20 (link)) and Papageorgiou et al. (21 (link)). Briefly, 50 μL extract, 450 μL distilled water and 2.5 mL 0.2 M Folin-Ciocalteu reagent were transferred into a test tube, vortexed (VTX-3000L Mixer Uzusio; LMS Co., Tokyo, Japan) for 1 min and allowed to stand for 8 min. Subsequently, 2 mL of an aqueous solution of Na₂CO₃ (75 g/L) were added and the mixture was left to react in the dark for 90 min at room temperature. The absorbance (relative to that of a blank prepared using methanol instead of extract) was measured at 765 nm using a double-beam UV-Vis spectrophotometer (model V-530; Jasco, Tokyo, Japan) and compared to the gallic acid calibration curve. The results were expressed in mg of gallic acid equivalents (GAE) per g of dry byproduct stream.

Tsouko E., Alexandri M., Fernandes K.V., Guimarães Freire D.M., Mallouchos A, & Koutinas A.A. (2019). Extraction of Phenolic Compounds from Palm Oil Processing Residues and Their Application as Antioxidants. Food Technology and Biotechnology, 57(1), 29-38.

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DPPH Antioxidant Assay of LSBE

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The DPPH antioxidant assay was performed according to an established method [68 (link)], modified as follows: LSBE was diluted with ethyl acetate (1:10, v/v), then 2 mL of a 10⁻⁴ M DPPH● stock solution previously prepared with ethyl acetate was added to 500 μL of diluted LSBE. An absorbance at 515 nm was read immediately after the addition of the (T0) and after 30 min of incubation (T30). The measurements were taken using a UV-VIS spectrophotometer, model V 530 (JASCO, Oklahoma City, OK, USA), and the results were expressed as inhibition percentage and calculated using the following formula: % inhibition = [(T0 − T30)/T0] × 100, where T0 represents the absorption at time zero and T30 represents the absorption at 30 min. The half maximum inhibitory concentration (IC50) was calculated using GraphPadPrism Software Version 8 (GraphPad Software, Inc. Avenida de la Playa La Jolla, San Diego, CA, USA), by linear regression analysis curve plotting between inhibition percentage and concentration of LSBE, zeaxanthin, and ascorbic acid. Ascorbic acid was used as the positive control. The DPPH assay was performed for six concentrations. Three technical replicates were used for each concentration. Data are presented as % DPPH inhibition of the three measurements ± SD (n = 3).

Visan S., Soritau O., Tatomir C., Baldasici O., Balacescu L., Balacescu O., Muntean P., Gherasim C, & Pintea A. (2023). The Bioactive Properties of Carotenoids from Lipophilic Sea buckthorn Extract (Hippophae rhamnoides L.) in Breast Cancer Cell Lines. Molecules, 28(11), 4486.

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Quantification of Residual Surfactant in Nanoparticles

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The residual surfactant in the NP matrix was evaluated by an already published colorimetric method [53 (link),54 (link)]. About 2 mg of NPs was solubilized in 0.5 mL of DCM and then added to 10 mL of water. After evaporation of DCM, the suspension was filtered through cellulose nitrate filter, porosity 0.45 m (Sartorius, Firenze, Italy), to remove the precipitated PLGA. Then, 2 mL of the aqueous solution was treated with 2 mL of 0.5% w/v BaCl₂ in HCl 1 N and 0.5 mL of I₂/KI (0.05 M/0.15 M) and incubated for 10 min in the dark. Poloxamer 188 concentration was calculated using a spectrophotometer (Model V530, Jasco, Cremella, Italy) measuring the absorbance at 540 nm, using a calibration curve from stock solutions of poloxamer 188 prepared under the same experimental conditions. Linearity was found in a range of 4–48 µg (R² = 0.9927). Due to the sensitivity to heat and light of the aqueous solution of I₂/KI, the calibration curve was calculated fresh before analysis. The analysis was performed in triplicate on three different NP formulations. The residual poloxamer 188 was calculated as follows:

Ottonelli I., Bighinati A., Adani E., Loll F., Caraffi R., Vandelli M.A., Boury F., Tosi G., Duskey J.T., Marigo V, & Ruozi B. (2022). Optimization of an Injectable Hydrogel Depot System for the Controlled Release of Retinal-Targeted Hybrid Nanoparticles. Pharmaceutics, 15(1), 25.

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Quantifying Residual Pluronic F68 in NMeds

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The residual amount of Pluronic^® F68 remaining in the formulated NMeds was evaluated using a previously published colorimetric method [67 (link)]. NMeds were solubilised in 0.5 mL of DCM and then slowly added to 10 mL of water. The organic solvent was evaporated by stirring at room temperature to precipitate the PLGA, which was then removed by filtration (cellulose nitrate filter, porosity 0.45 m, Sartorius, Firenze, Italy). An amount of 2 mL of the aqueous solution was treated with 2 mL of 0.5% w/v BaCl₂ in HCl 1 N and 0.5 mL of I₂/KI (0.05 M/0.15 M). This mixture was incubated for 10 min in the dark. Then, the Pluronic^® F68 concentration was calculated using a spectrophotometer (Model V530, Jasco, Cremella, Italy) measuring the absorbance at 540 nm, using a calibration curve made from stock solutions of Pluronic^® F68 prepared under the same experimental conditions. Linearity was found in the range of 4–48 µg/mL of Pluronic^® F68 (R² = 0.9927). Due to the sensitivity of the I₂/KI aqueous solution to heat and light, the standard curve was calculated fresh each day. The analysis was performed in triplicate on three different NMed formulations. The residual Pluronic^® F68 was calculated as follows:

Duskey J.T., Rinaldi A., Ottonelli I., Caraffi R., De Benedictis C.A., Sauer A.K., Tosi G., Vandelli M.A., Ruozi B, & Grabrucker A.M. (2022). Glioblastoma Multiforme Selective Nanomedicines for Improved Anti-Cancer Treatments. Pharmaceutics, 14(7), 1450.

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Determination of Total Polyphenols

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Total polyphenols were determined according to Singleton and Rossi (1965) . Frozen leaf samples were extracted with 100 mM sodium-phosphate buffer (pH 7) at a 1 : 4 (w/v) ratio using a mortar and pestle. Resulting extracts were centrifuged for 20 min, 14 000 g at 4 °C (Z 233 MK-2, HERMLE). Aliquots of the collected supernatant were assayed with 2 N Folin–Cioclateur reagent (Sigma) and 35 % Na₂CO₃, by 60-min incubation at 30 °C in a water bath. The absorbance of the resulting colour was measured at 730 nm (JASCO, model V-530) against a known standard of gallic acid. Total polyphenols were expressed in mg 100 g⁻¹ FW as gallic acid equivalents (GAE).

Ventura Y., Myrzabayeva M., Alikulov Z., Omarov R., Khozin-Goldberg I, & Sagi M. (2014). Effects of salinity on flowering, morphology, biomass accumulation and leaf metabolites in an edible halophyte. AoB Plants, 6, plu053.

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Optical Properties of Biopolymer Films

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All films were cut into rectangular pieces (10 mm × 30 mm) and placed on the internal side of a spectrophotometer cell. Transparency of films was measured by a spectrophotometer (Model V-530, Jasco International, Tokyo, Japan) at 600 nm. Five replicates of films were tested. The transparency (%) was calculated as the percentual relationship between the light intensity with the specimen in the beam and the light intensity with no specimen in the beam [26 (link)]. Color of films were determined with a colorimeter (Minolta chromameter CR-200) using the CIELAB color parameters. L, a, and b values were averaged from nine readings across for each sample on a white background: L = 83.96 ± 0.06; a = 2.05 ± 0.09; b = 1.99 ± 0.07. The total color difference (ΔE) was calculated according to Equation (1):
where ΔL*, Δa* and Δb* are the differentials between a sample color parameter and the color parameters of formulation without Col and without KS used as standard [27 (link)].

Liang X., Feng S., Ahmed S., Qin W, & Liu Y. (2019). Effect of Potassium Sorbate and Ultrasonic Treatment on the Properties of Fish Scale Collagen/Polyvinyl Alcohol Composite Film. Molecules, 24(13), 2363.

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DPPH Antioxidant Activity Assay

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The antioxidant activity of the methanolic extracts was assayed using the DPPH free radical scavenging method as proposed by Scherer and Godoy (22 (link)). Briefly, 100 μL of diluted (1:2, 1:5, 1:10, 1:20 V/V) methanolic extract were added to 3.9 mL DPPH solution (80.1 μM in methanol) and left in the dark at room temperature for 90 min. The absorbance was measured at 517 nm against methanol as a blank, using a double-beam UV-Vis spectrophotometer (model V-530; Jasco). A mixture consisting of 100 μL methanol and 3.9 mL DPPH solution was used as a control. The antioxidant activity index (AAI) was calculated as suggested and described thoroughly by Scherer and Godoy (22 (link)): where IC₅₀ is the concentration of the methanolic extract providing 50% inhibition.

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Photophysical Characterization of Photosensitizers

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UV-vis absorption spectra of all PSs in DMSO were recorded on an ultraviolet visible spectrophotometer (Jasco Model V-530, Japan) at wavelengths from 300 to 800 nm. Fluorescence spectra were carried out using a uorescence spectrometer (FluoroMax-4, France) in the range 500-800 nm using 425 nm excitation wavelength. All the measurements were carried out at room temperature. Cells were cultured in normal RPMI-1640 culture medium with 10% fetal bovine serum (FBS), 50 units per mL penicillin and 50 mg mL - 1 streptomycin in 5% CO 2 at 37℃.

Li M.Y., Mi L., Meerovich G., Soe T.W., Chen T., Than N.N., Yan Y.J, & Chen Z.L. (2022). The biological activities of 5,15-diaryl-10,20-dihalogeno porphyrins for photodynamic therapy. Journal of cancer research and clinical oncology, 148(9).

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Synthesis and Characterization of Porphyrin Derivatives

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Materials 5,15-Diaryl-10,20-dihalogeno porphyrins (I 1 - 6 , II 1 - 4 ) were synthesized in our laboratory (Fig. 2). All solvents and reagents were purchased from commercial suppliers and dichloromethane was used after further redistillation. HMME was donated by Shanghai Xianhui Pharmaceutical Co. Ltd. Thin-layer chromatography (TLC) analysis was carried out on silica gel plates GF254 and column chromatography was performed on silica gel (300-400 mesh). 1 H NMR and 13 C NMR spectra were recorded on a Bruker 400 MHz or 600 MHz spectrometer. MALDI-TOF mass spectra were recorded using a JEOL JMS-S3000 Spiral-TOFMS (JEOL, Tokyo, Japan). UV-vis absorption spectra were recorded on an UV-vis spectrophotometer (Jasco Model V-530, Japan). Fluorescence spectra were measured on a uorescence spectrophotometer (FluoroMax-4, France). The geometry optimization of PS 1 , I 2 , I 5 and I 6 in the DMSO was carried out by DFT calculations using the B3LYP functional with a 6-31G basis set for carbon, hydrogen, nitrogen, oxygen, chlorine, bromine atoms and LANL2DZ for iodine atoms. Graphics were prepared by OriginPro 9.0 and GraphPad Prism 8. All results were presented as mean ± SD. Comparison among the groups was determined by one-way ANOVA followed by Bonferroni correction for multiple testing. P < 0.05 was considered statistically signi cant.

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