QCT, LUT, and 3-O-MQ content into crude extract, nanoemulsions, and skin/mucosa after permeation studies were determined using a liquid chromatography equipment Shimadzu LC-10A, equipped with LC-10AD pump, CBM-10A system controller, SIL-10A autosampler, SPD-20AV UV/vis detector (set at 362 nm), and LC Solution software. The chromatographic system was composed by a Synergi Polar-RP 150 × 4.6 mm i.d., 4 μm (Phenomenex, Torrance, CA) column protected by precolumn packed with silica C18 Phenomenex (150 μm, 140 Å), temperature system of 30 ± 1°C, isocratic flux of 0.8 mL/min, and 20 μL as injection volume. The mobile phase consisted of methanol : 0.16 M phosphoric acid : acetonitrile (46 : 44 : 10, v/v/v) and the samples were diluted in methanol : phosphoric acid 16 mM (50 : 50, v/v) before analyses. The analytical method was previously validated for the determination of QCT, LUT, and 3-O-MQ in ethanolic extract and nanoemulsion [8 (link)], demonstrating to be specific, linear (0.25 to 10 µg/mL), precise, and accurate. In this paper, the revalidation of the analytical method was performed in terms of specificity and recovery of QCT, LUT, and 3-O-MQ from porcine ear skin and porcine esophageal mucosa. The limits considered acceptable for the evaluated parameters are in accordance with the “The Guidance for Industry: Bioanalytical Method Validation” (FDA).
Spd 20av uv vis detector
Manufactured by Shimadzu
Sourced in Japan, United States
The SPD-20AV UV/Vis detector is a laboratory equipment component designed to measure the absorbance of light by a sample. It is a key part of liquid chromatography systems, providing quantitative analysis of compounds by detecting their absorption of ultraviolet or visible light.
Automatically generated - may contain errors
Lab products found in correlation
Lc 20ad pump, by Shimadzu (3 mentions)
Cmb 20a system controller, by Shimadzu (3 mentions)
Sil 20ac autosampler, by Shimadzu (3 mentions)
Lc 20ad, by Shimadzu (2 mentions)
Pepsin, by Merck Group (2 mentions)
Pancreatin, by Merck Group (2 mentions)
Sil 20a autosampler, by Shimadzu (2 mentions)
Prominence chromatographic system, by Shimadzu (2 mentions)
Folin ciocalteau reagent, by Merck Group (2 mentions)
Trolox, by Merck Group (2 mentions)
Ferric chloride, by Lach-Ner (2 mentions)
Sodium nitrite, by Lach-Ner (2 mentions)
Lc 20at binary pump, by Shimadzu (2 mentions)
Multiskan go microplate reader, by Thermo Fisher Scientific (2 mentions)
Uv 1800 spectrophotometer, by Shimadzu (2 mentions)
Prominence hplc, by Shimadzu (2 mentions)
Sil 10a autosampler, by Shimadzu (1 mentions)
Cbm 10a system, by Shimadzu (1 mentions)
Lc 10ad pump, by Shimadzu (1 mentions)
Lc 10a, by Shimadzu (1 mentions)
16 protocols using spd 20av uv vis detector
1
Quantification of Flavonoids from Skin and Mucosa
2
Phenolic Compounds Analysis by HPLC
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Phenolic compounds were analyzed by high-performance liquid chromatography (HPLC) equipped with two UV/VIS detectors (SPD-10A UV/VIS Detector; Shimadzu SPD-20AV UV-Vis Detector, Shimadzu, Kyoto, Japan), a column oven (CTO 10AVP Column Oven, Shimadzu) and a reverse-phase C18 column (Prevail 5 µm organic acid, 4.6 mm × 250 mm, Hichrom, Leicestershire, UK). The mobile phase solution consisted of 2.5 mM KH2PO4 and acetonitrile (60:40 v/v). The flow rate applied was 1.0 mL/min. Then, 20 μL of the sample was injected at 40 °C with a run time of 20 min. Two UV/VIS detectors were used with detection wavelengths set to either 305 nm or 280 nm. Phenols in each sample were analyzed in triplicate.
Phenols of each compound were identified by comparing the retention time of the external standard with that of the internal standard. Phenolic standard solutions were prepared by dissolving in methanol or acetonitrile. A calibration curve was created by analyzing the standard solutions of each compound diluted to several concentrations. Quantification was performed using the peak area and the calibration curve of each compound.
Phenols of each compound were identified by comparing the retention time of the external standard with that of the internal standard. Phenolic standard solutions were prepared by dissolving in methanol or acetonitrile. A calibration curve was created by analyzing the standard solutions of each compound diluted to several concentrations. Quantification was performed using the peak area and the calibration curve of each compound.
3
HPLC Characterization of Compounds
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The HPLC instrumentation equipped with a vacuum degasser, a quaternary pump (LC-20AD, SHIMADZU), a column oven (CTO-20AC, SHIMADZU), and an auto-sampler (SIL-20AC, SHIMADZU), was connected to a SPD-20AV UV–Vis detector (SHIMADZU). Chromatographic analysis was performed on a Purospher star RP-C18 column (4.6 mm × 250 mm, 5 μm) by using a simultaneous elution and flow-rate programming RP-HPLC method.
4
Antioxidant Capacity Analysis of Whey Protein
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Folin-Ciocalteau reagent, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH●), β-carotene, pancreatin, pepsin, Trolox and trichloroacetic acid were purchased from Sigma Chemical Co. (St Louis, MO, USA), ferric chloride was obtained from J.T. Baker (Deventer, Holland), and sodium nitrite from LACH-NER (Brno, Czech Republic). Other chemicals and solvents were of the highest analytical grade. Whey protein isolate (WPI) was purchased from Olimp Laboratories (Debica, Poland). Distilled water was produced using water purification system DESA 0081 Water Still destilator (POBEL, Madrid, Spain). Absorbance in spectrophotometrical assays was measured on a Multiskan GO microplate reader (Thermo Fisher Scientific Inc., Waltham, MA, USA). For HPLC analysis a Shimadzu Prominence chromatographic system was used, which consisted of LC-20AT binary pump, CTO-20A thermostat and SIL-20A autosampler connected to the SPD-20AV UV/Vis detector (Shimadzu, Kyoto, Japan). Freeze-dryer, model Christ Alpha 2-4 LSC, was from Martin Christ (Osterode am Harz, Germany), and spray-dryer (Buchi mini B-290) from BüchiLabortechnik AG (Flawil, Switzerland).
5
SdFFF Separation Device Characterization
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The SdFFF separation device used in this study derived from previously described apparatuses [47 (link)] as schematized in Figure 3A . The apparatus was composed of two 880 x 47 x 2 mm polystyrene plates, separated by a Mylar® spacer in which the channel was carved. Channel dimensions were 788 × 12 × 0.175 mm with two 50 mm V-shaped ends. The measured total void volume (channel volume + connecting tubing + injection and detection device) was calculated after injection and retention time determination of a non-retained compound (0.10 g/L benzoic acid (Merck Millipore, Lyon, France) solution, UV detection at 254 nm). Sedimentation fields were calculated as previously described [48 (link)] and were expressed in gravity units (1 g = 980 cm/s2). Elution signals were recorded at 254 nm with a SPD-20AV UV/VIS Detector (Shimadzu, Champ sur Marne, France) and a NI9211 (10 mV input) acquisition device (National Instruments France, Nanterre, France) operated at 3 Hz and connected to a PC computer by the means of a laboratory-developed Visual Basic software (VB pro, Ver 6.0, Microsoft Corp.).
6
Comprehensive Phytochemical Analysis of Soy Protein
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Folin-Ciocalteau reagent, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH●), Trolox, trichloroacetic acid, pancreatin, pepsin and all standards for HPLC analysis (phenolics and betalains) were purchased from Sigma Chemical Co. (St Louis, MO, USA), ferric chloride was obtained from J.T. Baker (Deventer, Holland), and sodium nitrite from LACH-NER (Brno, Czech Republic). Other chemicals and solvents used were of the highest analytical grade. Distilled water was produced using water purification system DESA 0081 Water Still destilator (POBEL, Madrid, Spain). Soy protein isolate was purchased from “Macrobiotic Prom” company (Belgrade, Serbia). Absorbances in spectrophotometrical assays were measured on a Multiskan GO microplate reader (Thermo Fisher Scientific Inc., Waltham, MA, USA) and UV-1800 spectrophotometer (Shimadzu, Kyoto, Japan). For HPLC analysis a Shimadzu Prominence chromatographic system was used, which consisted of LC-20AT binary pump, CTO-20A thermostat and SIL-20A autosampler connected to the SPD-20AV UV/Vis detector (Shimadzu, Kyoto, Japan).
7
HPLC Analysis of [18F]DPA-714 Radiotracer
8
HPLC Analysis of Plant Extract
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A Shimadzu LC-20AD with SPD- 20AV UV-VIS Detector was used for the test. The mobile phase comprised 0.10 M acetic acid: acetonitrile using gradient elution. The percentages and times of varying the acetonitrile have been supplied in Table 8 . A solution of 10 mg/mL solution of PAE was constituted using methanol. A volume of 10 µL of the extract (10 mg/mL) was injected into the columns and allowed to run for 45 min at a flow rate of 0.5 mL/min, under a pump pressure of 1700 Psi and a temperature of 40°C. The chromatograms obtained were detected at a wavelength of 256 nm and the retention times, areas under curve, concentrations, heights, and areas (%) of the chromatograms were then determined.
9
Quantitative Analysis of Phenolic Compounds
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Total phenolic content (TPh) in the extracts was established using the Folin–Ciocalteau spectrophotometric method adapted to microscale [11 (link)]. The sample (15 μL) was mixed with distilled water (170 μL), 2 N Folin–Ciocalteu’s reagent (12 μL) and 20% Na2CO3 (30 μL) in a plate well. Absorbance was measured after 1 h (room temperature, dark conditions) at 750 nm, using distilled water as blank. Gallic acid (GAE) was used for the calibration curve.
Chromatographic analysis for identification and quantification of phenolic compounds were carried out as recommended by Tumbas Šaponjac et al. [11 (link)], using Shimadzu Prominence HPLC, connected to an SPD-20AV UV/VIS detector (Shimadzu, Kyoto, Japan), with Luna C-18 RP column, 5 lm, 250 mm × 4.6 mm with a C-18 guard column, 4 mm × 30 mm (Phenomenex, Torrance, CA, USA). Gradient elution was applied using acetonitrile (A) and water acidified with 1% formic acid in d-water (B), at flow rates of 1 mL/min, at the following order: 10% to 25% A (0–10 min); 25% to 60% A (10–20 min); 60% to 70% A (20–30 min); 70% to 10% A (30–40 min); 10% A (5 min) (equilibration time). For hydroxybenzoic acids, chromatograms were recorded at 280 nm; for hydroxycinnamic acids at 320 nm; and for flavonoids at 360 nm. HPLC standards were dissolved in 50% methanol.
Chromatographic analysis for identification and quantification of phenolic compounds were carried out as recommended by Tumbas Šaponjac et al. [11 (link)], using Shimadzu Prominence HPLC, connected to an SPD-20AV UV/VIS detector (Shimadzu, Kyoto, Japan), with Luna C-18 RP column, 5 lm, 250 mm × 4.6 mm with a C-18 guard column, 4 mm × 30 mm (Phenomenex, Torrance, CA, USA). Gradient elution was applied using acetonitrile (A) and water acidified with 1% formic acid in d-water (B), at flow rates of 1 mL/min, at the following order: 10% to 25% A (0–10 min); 25% to 60% A (10–20 min); 60% to 70% A (20–30 min); 70% to 10% A (30–40 min); 10% A (5 min) (equilibration time). For hydroxybenzoic acids, chromatograms were recorded at 280 nm; for hydroxycinnamic acids at 320 nm; and for flavonoids at 360 nm. HPLC standards were dissolved in 50% methanol.
10
Characterization of Carbon Black Nanoparticles
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The AF4 system used was an AF2000 MultiFlow FFF (Postnova Analytics GmbH, Landsberg am Lech, Germany) coupled online to an SPD-20AV UV–Vis detector (Shimadzu Corporation, Japan) and a Zetasizer Nano ZS DLS detector (Malvern, UK). Flows were provided by two separate PN1130 isocratic pumps (Postnova Analytics GmbH) equipped with a PN7520 degasser, and the cross-flow was obtained by a separate piston pump, which is constantly adjustable. The DLS detector can work in batch mode as well. Table S6 and S7 of the SI summarize the optimal conditions for batch DLS measurements and the optimal instrumental variables and conditions of the AF4 system for the study of CB-NP dispersions, respectively.
The morphology was studied by scanning electron microscopy (SEM) with a Hitachi S-4800 instrument at an accelerating voltage of 10.0 keV over metallized CB solid samples with a mixture of gold and palladium for 30 s. Transmission electron microscopy (TEM) samples were prepared by delivering 10 μL of the CB dispersion onto a carbon-coated copper grid (300 mesh) and were dried overnight at room temperature. These samples were analysed using a JEM-1010 (JEOL Ltd.) operated at 100 kV.
The morphology was studied by scanning electron microscopy (SEM) with a Hitachi S-4800 instrument at an accelerating voltage of 10.0 keV over metallized CB solid samples with a mixture of gold and palladium for 30 s. Transmission electron microscopy (TEM) samples were prepared by delivering 10 μL of the CB dispersion onto a carbon-coated copper grid (300 mesh) and were dried overnight at room temperature. These samples were analysed using a JEM-1010 (JEOL Ltd.) operated at 100 kV.
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