HPLC analyses were performed on a Shimadzu Prominence series HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a solvent reservoir, a prominence degasser (DGU-20A3), a prominence pump (LC-20AD), prominence auto-sampler (SIL-20A HT), a column oven (CTO-10AS VP), a prominence UV/VIS detector (SPD-20A), and computer software LabSolutions® Version 5.30 SP1, 2010.
Prominence series hplc system
Manufactured by Shimadzu
Sourced in Japan
The Prominence series HPLC system from Shimadzu is a high-performance liquid chromatography system designed for analytical purposes. It features a modular design with various components, including pumps, autosampler, column oven, and detectors, allowing for customization to meet specific analytical requirements.
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Lab products found in correlation
Lc 20ab binary pump, by Shimadzu (4 mentions)
Sil 20a ht autosampler, by Shimadzu (4 mentions)
Hplc grade, by Thermo Fisher Scientific (4 mentions)
Cto 20ac temperature controlled column oven, by Shimadzu (3 mentions)
Cbm 20a communications bus, by Shimadzu (3 mentions)
Protein sequencing grade, by Merck Group (2 mentions)
Labsolutions lite software version 5.71 sp2, by Shimadzu (2 mentions)
Trifluoroacetic acid, by Merck Group (2 mentions)
Spd m20a photodiode array detector, by Shimadzu (2 mentions)
Lab solutions lite software version, by Shimadzu (2 mentions)
Dgu 20a3, by Shimadzu (1 mentions)
Spd 20a, by Shimadzu (1 mentions)
Lc 20ad, by Shimadzu (1 mentions)
Lc ms grade, by Thermo Fisher Scientific (1 mentions)
Tskgel ods 100v, by Tosoh (1 mentions)
Cto 20ac, by Shimadzu (1 mentions)
Spd m20a, by Shimadzu (1 mentions)
7 protocols using prominence series hplc system
1
HPLC Analysis of Chemical Compounds
2
HPLC Characterization of Dynantin and MDP
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All analyses were performed using a Shimadzu Prominence series HPLC system (Shimadzu Corporation, Kyoto, Japan), equipped with a LC-20AB binary pump (Serial: L20124200883), SIL-20A HT autosampler (Serial: L20345256104), CTO-20AC temperature controlled column oven (Serial: L2021525077), and CBM-20A communications bus (Serial: L20235154327). All equipment was controlled by Shimadzu LabSolutions Lite software version 5.71 SP2. For separation, an Ultra C18 column, 3 μm, 50 x 4.6 mm (RESTEK Corporation, Bellefonte, PA) was used.
Dynantin samples were analyzed at a constant solvent flow rate of 0.7 mL/min at 35 °C using a binary gradient (Table 1 ). Solvent A consisted of a 25% solution of acetonitrile (HPLC grade Fisher Scientific) in ddH2O (0.2 μm filtered) and solvent B consisted of acetonitrile, with each solvent containing 0.1% trifluoroacetic acid (v/v, protein sequencing grade, Sigma Aldrich, Fairlawn NJ).
MDP samples were analyzed at a constant solvent flow rate of 1.0 mL/min at 35 °C using a binary gradient (Table 2 ). Solvent A consisted of ddH2O (0.2 μm filtered) and solvent B consisted of methanol (MeOH) (HPLC grade, Fisher Scientific, Fairlawn NJ), with each solvent containing 0.1% formic acid (v/v, LC/MS grade, Fisher Scientific).
Dynantin samples were analyzed at a constant solvent flow rate of 0.7 mL/min at 35 °C using a binary gradient (
MDP samples were analyzed at a constant solvent flow rate of 1.0 mL/min at 35 °C using a binary gradient (
3
HPLC Analysis of Dynantin Compounds
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All analyses were performed using a Shimadzu Prominence series HPLC system (Shimadzu Corporation, Kyoto, Japan), equipped with an LC-20AB binary pump (Serial: L20124200883), SIL-20A HT autosampler (Serial: L20345256104), CTO-20AC temperature-controlled column oven (Serial: L2021525077), SPD-M20A photodiode array detector and CBM-20A communications bus (Serial: L20235154327). All equipment were controlled by Shimadzu Lab Solutions Lite software version 5.71 SP2. For separation, an Ultra C18 column, 3 μm, 50 × 4.6 mm (RESTEK Corporation, Bellefonte, PA) was used. Dynantin samples were analyzed at a constant solvent flow rate of 0.7 mL/min at 35 °C using a binary gradient (Table 1 ). Solvent A consisted of a 25% solution of acetonitrile (HPLC grade, Fisher Scientific, Fairlawn, NJ, USA) in ddH2O (0.2 μm filtered) and solvent B consisted of acetonitrile with each solvent containing 0.1% trifluoroacetic acid (v/v, protein sequencing grade, Sigma Aldrich, Fairlawn, NJ, USA).
4
Radiochemical Quantification of Nucleoside Phosphorylation
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Ten micromolar tritium-labeled nucleosides and nucleoside analogs were incubated for 5 min at 37°C in 100-μl reactions containing 150 mmol/l Tris-HCl (pH 7.6), 6 mmol/l ATP, 15 mmol/l MgCl2, 1% bovine serum albumin (BSA) and 20 ng recombinant human TK1. Reactions were stopped by the addition of 200 μl of methanol and clarified by ultrafiltration. After clarification, solutions were desiccated by exposure to N2 gas, dissolved in 100 μl H2O, and analyzed by radio-high-performance liquid chromatography (HPLC). HPLC was performed using a 150TR Flow System analyzer (Perkin-Elmer) and a Prominence Series HPLC system (Shimadzu Corp., Kyoto, Japan) equipped with an ODS reverse-phase column (TSKgel ODS-100V, 250 mm × 4.6 mm, 3 μm; Tosoh Corp., Tokyo, Japan), with the eluent [10 mmol/l phosphate buffer (pH 3.0):Acetonitrile = 9:1] set to a flow rate 1.0 ml/min for 15 min. This procedure enabled quantitation of residual nucleosides and the production of nucleoside monophosphates formed in the reactions just described.
5
HPLC Analysis of Dynantin Compound
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All analyses were performed using a Shimadzu Prominence series HPLC system (Shimadzu Corporation, Kyoto, Japan), equipped with a LC-20AB binary pump, SIL-20A HT autosampler, CTO-20AC temperature-controlled column oven, SPD-M20A photodiode array detector, and CBM-20A communications bus. All equipment was controlled by Shimadzu Lab Solutions Lite software version 5.71 SP2. For separation, an Ultra C18 column, 3 μm, 50 mm × 4.6 mm (RESTEK Corporation, Bellefonte, PA, USA) was used. Dynantin samples were analyzed at 35 °C and a constant solvent flow rate of 0.7 mL/min using a binary gradient (Table 2 ). Solvent A consisted of a 25% solution of acetonitrile (HPLC grade, Fisher Scientific, Fairlawn, NJ, USA) in ddH2O (0.2 μm filtered) and solvent B consisted of acetonitrile with each solvent containing 0.1% trifluoroacetic acid (v/v, protein sequencing grade, Sigma Aldrich, Fairlawn, NJ, USA).
6
Quantification of Stigmasterol and Quercetin
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HPLC was used to determine the quantity of stigmasterol and quercetin at 258 nm and 206 nm, respectively. The experiments were carried out on a Shimadzu Prominence series HPLC system, which included a solvent delivery unit (model LC-20AD), a variable wavelength UV/VIS detector (model SPD-20A), and a Hamilton manual injection valve with a 20 μl loop, C18 column (250 × 4.6 mm, 5 m i.d.), and degasser (Aczet, Helix Biosciences). LC solution software was used for data collection and analysis. Methanol/0.1 percent formic acid (65 : 35 v/v) was employed as the mobile phase for quercetin, while methanol was used for stigmasterol. Calibration curve was prepared by using standard quercetin and stigmasterol.
7
HPLC Analysis of Compounds
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All analyses were performed using a Shimadzu Prominence series HPLC system (Shimadzu Corporation, Kyoto, Japan), equipped with a LC-20AB binary pump (Serial: L20124200883), SIL-20A HT autosampler (Serial: L20345256104), CTO-20AC temperature controlled column oven (Serial: L2021525077), SPD-M20A photodiode array detector and CBM-20A communications bus (Serial: L20235154327). All equipment was controlled by Shimadzu Lab Solutions Lite software version 5.71 SP2. For separation, an Ultra C18 column, 3 μm, 150 × 4.6 mm (RESTEK Corporation, Bellefonte, PA, USA) was used. Samples were analyzed at a constant solvent flow rate of 1.0 mL/min at 35 °C using a binary gradient (Table 1 ). Solvent A consisted of a ddH2O (0.2 μm filtered) and solvent B consisted of 20% ddH2O in acetonitrile (HPLC grade, Fisher Scientific, Fairlawn, NJ, USA) with each solvent containing 0.18% formic acid (v/v, LC/MS grade) and 0.15% triethylamine (v/v, HPLC grade, Fisher Scientific, Fairlawn, NJ, USA).
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