The quartz slides were washed by sonication in petroleum ethyl ether and methanol, dried in a vacuum oven and exposed to air plasma (Harrick, Plasma cleaner/sterilizer) for 10 min. The cleaned quartz slides were silanized by APTES according to Krishnan et al.[33 (link)] The hydrophobized quartz slide was coated with fluorescently labelled albumin (AlbFITC) and Hep layers using the procedure described in the section 2.1. Alb was fluorescently labelled with FITC (AlbFITC) to enhance absorbance at 280 nm. After each AlbFITC/Hep bilayer deposition, the UV/Vis absorption spectrum (Specord Plus, Analytik Jena) in the range of 200–550 nm was measured.
Specord plus
Manufactured by Analytik Jena
Sourced in Germany
The Specord Plus is a high-performance UV-Vis spectrophotometer designed for precise and accurate measurements of absorption, transmission, and reflection spectra. It features a compact design, intuitive user interface, and advanced optical components to deliver reliable results across a wide range of applications.
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Lab products found in correlation
Plasma cleaner sterilizer, by Harrick (1 mentions)
Avance 3 500 mhz, by Bruker (1 mentions)
Flash 2000 chns o analyzer, by Thermo Fisher Scientific (1 mentions)
Cp 505 ph meter, by Elmetron (1 mentions)
Multiskan fc microplate photometer, by Thermo Fisher Scientific (1 mentions)
Rapamycin, by Abcam (1 mentions)
Nocodazole, by Merck Group (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Synapt g2 si, by Waters Corporation (1 mentions)
Cisplatin, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Sensline avaspec 2048xl, by Avantes (1 mentions)
Quanta 200, by Thermo Fisher Scientific (1 mentions)
Avance 400, by Bruker (1 mentions)
12 protocols using specord plus
1
Silanized Quartz Slides for Protein Immobilization
2
Synthesis and Characterization of Schiff Base
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All chemicals were obtained from the Aldrich (Saint Louis, MO, USA), TEDIA (Fairfield, OH, USA), and Merck (Rahway, NJ, USA) companies. The melting point of the synthesized Schiff base was determined by employing an electrothermal melting point SMP 10 apparatus. The FTIR spectrum was detected using an FTIR Bruker-ATR (Leipzig, Germany). A Bruker Avance III −500 MHz was used to record the 1H and 13C NMR spectra in DMSO solvent. The mass spectra were recorded on a Brucker apex-IV. The UV–visible spectrum of the Schiff base–DMSO solution was recorded in the range from 200 to 900 nm using a SPECORD PLUS by Analytik Jena AG. GCMS ISQ models from Thermo Scientific were used for direct inlet mass spectrometry, and the FLASH 2000 CHNS/O analyzer was used to determine the elemental contents of the synthesized compounds.
3
Spectrophotometric Quantification of Functionalized Polymers
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Spectrophotometric analysis of functionalized polymers was carried out in quartz glass cuvettes on a Specord Plus UV–VIS spectrophotometer (Analytik Jena, Germany). The molar content of DTB and TT groups in the polymers was determined at 302 and 305 nm in methanol using the molar absorption coefficient of 12 100 and 10 300 L mol−1 cm−1, respectively26 (link). DFA group content was measured indirectly at 432 nm upon addition of FeCl3∙6H2O (2 eq. compared to the theoretical content of DFA groups) to the aqueous solution of polymers using a molar absorption coefficient of 2 585 L mol−1 cm−127 (link). The resulting values of functional group contents were obtained by arithmetic average of 3 independent measurements.
4
Quantification of Phenolic Compounds and Free Amino Acids
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A CP-505 pH meter (Elmetron, Zabrze, Poland) was used for pH measurement. Total polyphenols were determined by the Folin-Ciocalteu method and spectrophotometric measurement at 670 nm (Singleton & Rossi, 1965) . Total procyanidins were determined by the vanillin method following the procedure previously described (Monagas, Gómez-Cordovés, & Bartolomé, 2006) . Neutral polysaccharides were determined by the phenol/sulfuric acid method according to a previous procedure (Segarra, Lao, López-Tamames, & De La Torre-Boronat, 1995) . Free amino acids were determined by method 5 of Doi and co-workers (Doi, Shibata, & Matoba, 1981) A Multiskan™ FC Microplate Photometer (Thermo Fisher Scientific, Massachusetts, USA) and a Specord ® plus (Analytik Jena AG, Jena, Germany) spectrophotometer were used for these determinations.
5
Determination of Intestinal Enzyme Activities
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The determination of the pig intestinal β-galactosidase activity was adapted from Warmerdam et al. (2014). 29 Briefly, a solution of o-NPG (0.5 mg/mL) in phosphate buffer 0.05 M, pH 7.0 was prepared. The enzymatic activity was determined by incubating 1,900 μL of the o-NPG solution and 100 μL of enzyme solution from BBMV for 2 h at 37 °C.
The method is based on the measurement of the continuous release of o-NP from o-NPG.
The absorbance of released o-NP was measured at 420 nm every 30 s using a spectrophotometer (Specord Plus, Analytik Jena) together with a temperature controller (Jumo dTRON 308, Jumo Instrument Co.). The specific enzymatic activity (U) was expressed in μmol min -1 g -1 , where one unit was defined as the amount of enzyme that produced 1 μmol of o-NP in one min of reaction (n = 3). Similar procedure was used to determine the maltase activity by using a solution of p-NPG in phosphate buffer 0.05 M, pH 6.8 (0.05% w/w) and monitoring the release of p-NP at 420 nm every 20 s (n = 3).
The method is based on the measurement of the continuous release of o-NP from o-NPG.
The absorbance of released o-NP was measured at 420 nm every 30 s using a spectrophotometer (Specord Plus, Analytik Jena) together with a temperature controller (Jumo dTRON 308, Jumo Instrument Co.). The specific enzymatic activity (U) was expressed in μmol min -1 g -1 , where one unit was defined as the amount of enzyme that produced 1 μmol of o-NP in one min of reaction (n = 3). Similar procedure was used to determine the maltase activity by using a solution of p-NPG in phosphate buffer 0.05 M, pH 6.8 (0.05% w/w) and monitoring the release of p-NP at 420 nm every 20 s (n = 3).
6
Synthesis and Characterization of Anticancer Compounds
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All commercial reagents and solvents were used without further purification. Cisplatin is purchased from Alfa Aesar (Heysham, UK); rapamycin from Abcam (Cambridge, UK); doxorubicin, nocodazole and DMSO from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Stock solutions were prepared in DMSO. Melting points were determined with a Barnstead Electrothermal (BI 9300) capillary melting point apparatus and are uncorrected. Elemental analyses were performed with a varioMICRO analyser. Thin layer chromatography (TLC) was carried out on aluminium-baked (Macherey-Nagel GmbH, Düren, Germany) silica gel 60. Column chromatography was performed on silica gel (230-400 mesh). The electronic absorption spectra were acquired on a UV-Vis double beam spectrophotometer SPECORD® PLUS (Analytik Jena GmbH, Germany). The molar conductance measurement was carried out using a CDRV 62 Tacussel electronic bridge, employing a calibrated 10-2 M KCl solution and 10-3 M solutions of compounds in DMSO. Purities of all tested compounds were ≥95%, as estimated by HPLC analysis. High Resolution Mass Spectrum (HR-MS) was measured at REALCAT (Université de Lille) on a Synapt G2Si (Waters) equipped with an ion mobility cell.
7
Spectrophotometric Analysis of Porcine Hemoglobin and Melanin
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To measure the relative absorbance of porcine hemoglobin and melanin the samples are diluted in distilled water and put in a 1 ml UV-cuvette, respectively. As reference 1 ml pure distilled water is used. The absorbance is measured using a spectrometer (SPECORD PLUS of Analytik Jena, Jena (Germany)) in the wavelength range between 350 and 1100 nm.
8
Quantification of Protein Content in RSIE
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The total protein content of RSIE was quantified by the Bradford method, using the Bio-Rad protein assay kit and bovine serum albumin as a standard [36 (link)]. Finally, the absorbance was monitored at 595 nm using a spectrophotometer (Specord Plus, Analytik Jena) together with a temperature controller (Jumo dTRON 308, Jumo Instrument Co). The protein content of RSIE was 7.5 ± 0.6% (w/w).
9
Antioxidant Analysis of Biopolymer Films
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The antioxidant properties of the films were analyzed through the DPPH assay. To conduct this analysis, the film was dissolved in 5 mL of ethanol, and then 3 mL of a DPPH solution (0.008% w/v in ethanol) was introduced into the film solution and mixed well. The resulting mixture was kept in the dark for 30 min. Subsequently, the absorbance value at a wavelength of 517 nm was measured using a UV-visible spectrophotometer (SPECORD PLUS, Analytik Jena, Jena, Germany). The DPPH radical scavenging activity (AA) of the films was then calculated as follows:
where ADPPH is the absorbance of the control, and Asample is the absorbance of the sample.
where ADPPH is the absorbance of the control, and Asample is the absorbance of the sample.
10
Physicochemical Characterization of Nano-Emulsions
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Physiochemical properties of the prepared NE were determined by using standard procedures and results are reported as average of three readings. Refractive index was measured directly (without dilution) of all the NE and NS-1 using an Abbes type refractometer (Sigma Instruments, LI- AREF-186) at 25 ± 1 °C (Tubesha et al. 2013 ). Transmittance (%), of the NE and NS was determined by using UV–vis spectrophotometer (Analytikjena, SPECORD plus), after preparing 1 % solution in distilled water and taking absorbance at 250 nm. Viscosity and Density of undiluted NE and NS-1 was determined using Ostwald viscometer (Fisher scientific, SI Analytics™ 285404014) and pycnometer while maintaining temperature at 25 ± 1 °C.
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