The HPLC analysis of brusatol in BE-BJO and BF-BJO was performed according to our previous investigation [17 (link)]. Measurements were carried out using HPLC LC-20AT (Shimadzu, Kyoto, Japan). PRONTOSIL120-3-C18-ace-EPS column (2.0 mm × 50 mm, 3.0 μm, Bischoff, Germany). The injection volume was 10 μL, the detection wavelength was 254 nm, and the he flow rate was set at 0.5 mL/min.
Lc 20at hplc
Manufactured by Shimadzu
Sourced in Japan, China
The LC-20AT HPLC is a high-performance liquid chromatography system manufactured by Shimadzu. It is designed to perform liquid chromatography analysis, separating and detecting various chemical compounds in a sample.
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Lab products found in correlation
Q exactive, by Shimadzu (1 mentions)
C18 column, by Shimadzu (1 mentions)
Gcms qp2020, by Shimadzu (1 mentions)
Gc 2014c, by Shimadzu (1 mentions)
1100 sl mass spectrometer, by Agilent Technologies (1 mentions)
Avance 400, by Bruker (1 mentions)
Nova 60, by Merck Group (1 mentions)
Dr3900, by HACH (1 mentions)
Spd 20a uv, by Shimadzu (1 mentions)
Cto 20a column oven, by Shimadzu (1 mentions)
Lc 20at pump, by Shimadzu (1 mentions)
Hi 2221, by Hanna Instruments (1 mentions)
Rid 20a detector, by Shimadzu (1 mentions)
Lc 20at system, by Shimadzu (1 mentions)
Venusil innova durashell nh2 column, by Agela Technologies (1 mentions)
Welch ultimate xb c18 column, by Hill-Rom (1 mentions)
Jem 1400plus, by JEOL (1 mentions)
Eclipse plus c18 column, by Agilent Technologies (1 mentions)
Spd m20a, by Shimadzu (1 mentions)
Toc vcsh analyzer, by Shimadzu (1 mentions)
28 protocols using lc 20at hplc
1
HPLC Analysis of Brusatol in BE-BJO and BF-BJO
2
Photocatalytic Oxidation and H2 Evolution
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Photocatalytic oxidation of 1-phenylethanol integrated with hydrogen (H2) evolution was conducted in a 40-ml sealed quartz reactor [42 ]. In a typical experiment, 5 mg of catalyst was dispersed in 5 ml of CH3CN containing 1-phenylethanol (0.1 mmol). The solution was purged with Ar for 20 min to expel the air inside and then illuminated with UV-vis light (300 nm ≤ λ ≤ 800 nm) through a 300-W Xe arc lamp (PLS-SXE 300D, Beijing Perfectlight Co., Ltd) for 2 h at room temperature. The incident light intensity was 300 mW·cm−2 which was identified by a PL-MW2000 photoradiometer (Beijing Perfectlight Co., Ltd). The release of H2 was quantified via a gas chromatograph (Shimadzu GC-2014C, 5A column, Ar carrier). The liquid products were analyzed by HPLC (Shimadzu HPLC-LC20AT, C18 column) and GC-MS (Shimadzu GC-MS QP 2020, Q-Exactive).
3
Solvent-shift Supersaturation and Stabilization
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A solvent-shift method was employed to generate FNB supersaturation.24 (link) The apparent drug concentration–time profile and the duration of the supersaturated state were determined using a USP II paddle method with a ZRS-8G dissolution tester (TDTF, Tianjing, China). A total medium volume of 200 mL with or without 0.05% (w/w) predis-solved polymer was maintained at 37°C±0.5°C and the stirring speed was fixed at 100 rpm. The polymers used to stabilize the supersaturation were HPMC E3, HPMC E5, HPMC E15, PVP K30, PVP K90, PVP VA 64, and Soluplus. After adding 200 μL of FNB stock solution (100 mg/mL) in dimethylsulfoxide to the test medium, 2 mL of samples were withdrawn without volume replacement at 10, 30, 60, 90, and 120 min and filtered through a 0.22 μm filter. All filters were prerinsed with 5 mL of a saturated solution of FNB in order to prevent adsorption onto the filter. The resulting filtrate was diluted appropriately in absolute ethanol and assayed by high-performance liquid chromatography (HPLC LC-20AT; Shimadzu, Kyoto, Japan) at a wavelength of 286 nm. The area under the concentration–time curve in supersaturated state (AUCss) was computed according to the linear trapezoidal rule. Each dissolution test was performed in triplicate.
4
Synthesis and Characterization of Novel Organic Compound
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Unless otherwise noted, all reagents and solvents were obtained from commercially available sources and were used without purification. Reactions were monitored by TLC with silica gel plates under UV light (254 and 365 nm). The melting point was measured (uncorrected) on a hot-stage microscope (Beijing Taike, X-4). 1H NMR and 13 C NMR spectra were tested in CDCl3 with TMS as the internal reference on a Bruker AVANCE 400 or 600 (1H at 400 or 600 MHz, 13 C at 150 MHz). Mass spectra (MS) were obtained from Agilent Co. Ltd. on an Agilent 1100-sl mass spectrometer with an electrospray ionisation source. HPLC (HPLC-LC-20AT, Shimadzu) using SPD-20A UV as the detector, C18 (4.6 × 250 mm, 0.45 mm) as HPLC analysis column. Method: gradient: 45% methanol in water as mobile elution, flow rate: 1.0 ml min−1.
5
Phenol Quantification by HPLC and Spectroscopy
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Phenol concentrations were measured using high-performance liquid chromatography HPLC LC-20AT (Shimadzu, Japan) equipped with a 4.6 mm reversed-phase C18 column (Phenomenex, Netherlands) and a UV detector at a wavelength of 280 nm. The mobile phase used was 25% (v/v) acetonitrile at a flow rate of 0.95 mL.min–1. The column oven was set at 30°C. Fast phenol measurements were also carried out by Merck – Spectroquant Phenol cell kits by using a spectrophotometer NOVA60 (Merck, Germany). Hach Lange kits were used to measure chemical oxygen demand (COD). The COD was measured using a VIS – spectrophotometer (DR3900, Hach Lange, Germany) making proper dilutions to minimize interference by high chloride concentrations, without compromising the accuracy of the measurement.
6
Redox-Responsive Biotin Release Kinetics
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The release of Deac from Deac-SS-Biotin was studied in PBS (pH 7.2–7.4) containing 20% DMSO with 0, 5, 10, or 20 μM, dithiothreitol (DTT), respectively. Briefly, biotin-conjugates (5 μM) were incubated in 10 ml of release medium at 37 °C. At timed intervals, 0.1 ml of sample was withdrawn, and the content of Deac, Deac-Biotin, and Deac-SS-Biotin was determined by HPLC (HPLC-LC-20AT, Shimadzu using SPD-20A UV as the detector, C18 (4.6 × 250 mm, 0.45 mm) as HPLC analysis column. Method: gradient: 45% methanol in water as mobile elution, flow rate: 1.0 ml min-1). The UV detector was kept at 254 nm.
7
Extraction and HPLC Analysis of Bacterial Metabolites
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The strains were inoculated into 100 mL marine agar, cultured at 28 °C for 72 h, centrifuged, and the bacteria were collected. Add 3 mL sterile water to the tube and wash the bacteria. Transfer the bacteria to a 15 mL centrifuge tube. Centrifuge at 4 °C, 8000×g for 10 min, then discard supernatant. The bacteria were resuspended with 2 mL acetone solution and extracted by shock for 10 min. After centrifugation at 8000×g for 10 min, the supernatant extract was transferred to a new tube. 2 mL ethyl acetate solution was added to the original tube, and the extraction was continued with oscillation for 10 min. After centrifugation at 8000×g for 10 min, the supernatant extract was mixed with the previous extract, and 3 mL sterile water was added to it, and the mixture was shaken and mixed. Centrifuge 8000×g for 10 min to delaminate the liquid and draw the upper liquid into a new EP tube. The ethyl acetate was dried to obtain powder and was redissolved with methanol. Over 0.22 µm filter membrane, used for High-pressure liquid chromatography (Shimadzu HPLC LC-20AT) analysis and detection. The tests were by C18 column with a mobile phase of methanol–acetonitrile–water (volume ratio being 80:15:5). UV detection was performed at 478 nm.
8
Quantifying Glucose and Fructose in Tej
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The concentrations of glucose and fructose in the samples were determined using a High-Performance Liquid Chromatography-Evaporative Light Scattering Detector (HPLC-ELSD) system63 (link). After centrifuging 5 mL Tej sample at 3200 rpm for 30 min, the supernatant was double filtered through 0.45 µm and 0.22 µm pore size membranes. Analyses were performed using an LC-20AT HPLC (Shimadzu, Japan), a YMC-Pack Polyamine II (250 4.6 mm. D.S-5 m, 12 nm) column, and an ELSD-LT II detector. At a column temperature of 30 °C, a 10 μL aliquot of the samples was injected into the HPLC-ELSD system. A mobile phase of acetonitrile: water at a 75:25 ratio with an isocratic pressure and a flow rate of 1.0 mL/min was used during the analysis period. Nitrogen (N2) was used as the nebulizer gas with a flow rate of 2 L/min. Finally, sugar quantification was performed by comparing the sugar peak areas of the samples to the corresponding standards calibration curves.
9
Quantification of β-Cyclocitral by HPLC
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The β-CY concentration was determined as previously described (Liu et al. 2014 (link); Zhao et al. 2015 (link)). In detail, 5 mL of LB-CY medium and acetonitrile were transferred into a 100 mL flask and shaken for 30 s by a vortex mixer. Then, the flask was under ultrasonic (40 kHz and 300 W) for 30 min (Liu et al. 2012 (link)). After the mixture was centrifuged at 8000 rpm for 20 min, the supernatant was collected and filtered through a 0.22 μm membrane filter. Finally, the obtained filtrate was used to determine its β-CY content by high-performance liquid chromatograph (HPLC). The content of β-CY was determined by LC-20AT HPLC (Shimadzu, Kyoto, Japan) equipped with LC-20AT pump (Shimadzu), a CTO-20A column oven (Shimadzu), a Kromasil C18 column (250 mm × 4.60 mm, 5.0 μm; Sweden) and an SPD-M20A detector. Degradation of β-CY was calculated according to the following equation: where C and C0 are the concentration of β-CY (mg/L) in inoculated medium and the control, respectively.
10
HPLC Analysis of Chemical Compounds
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The sample concentration is too high for liquid chromatography analysis, so samples were diluted 10-fold and analyzed by Shimadzu LC-20AT HPLC (Shimadzu Corporation, Kyoto, Japan) on Welchrom C18 columns (4.6 × 250 mm, 5 μm; Shanghai Yuexu Material Technology Co., Ltd., China). Detection conditions were as follows: wavelength 270 nm, column temperature was 25°C, mobile phase flow rate 1 ml min−1, injection volume 10 μL. The mobile phase was 0.2% aqueous phosphoric acid and methanol, adopting a gradient elution program of 5% of B at 0–6 min, 5%–7% of B at 6–15 min, 7%–15% of B at 15–20 min, 15%–21% of B at 20–25 min, 21%–22% of B at 25–41 min, 22%–28% of B at 41–47 min, 28%–32% of B at 47–55 min, 32%–37% of B at 55–61 min, 37%–38% of B at 61–62 min, 38%–39% of B at 62–67 min, 39%–45% of B at 67–70 min, 45%–65% of B at 70–80 min, 65%–5% of B at 80–90 min (Huang et al., 2018 (link)).
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