HPLC analyses were carried out with an Agilent 1260 Infinity Quaternary HPLC system (Santa Clara, CA, USA) equipped with a pump (G1312C) with an integrated degasser (G1322A), a thermostatted column compartment (G1316A), an autosampler (G1329B), a diode-array detector (G1315D), and online analysis software (Chemstation). Analyses were performed at 25 °C using a normal-phase Zorbax Sil (5 µm, 4.6 × 150 mm) column (Agilent Technologies, Santa Clara, CA) protected with a guard column. Carotenoid and retinal separation was achieved using an isocratic composition of 70:30 (v:v) of hexane: ethyl acetate. The flow rate for all systems was 1.4 ml/min. Detection of carotenoids and apocarotenoids was performed at 420 nm wavelength. For LC-MS analyses, the eluate was directed into a LXQ linear ion trap mass spectrometer (Thermo Scientific) through atmospheric pressure chemical ionization (APCI) source working in the positive mode. To ensure optimal sensitivity, the instrument was tuned with Z as well as with retinal standards.
1260 infinity quaternary hplc system
Manufactured by Agilent Technologies
Sourced in United States
The 1260 Infinity Quaternary HPLC system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative applications. It features a quaternary solvent delivery system, which allows for the simultaneous use of up to four solvents. The system is capable of delivering precise and accurate flow rates, enabling reliable and reproducible chromatographic separations.
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Lab products found in correlation
Lxq linear ion trap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
0.45 μm membrane filter, by Merck Group (1 mentions)
Zorbax eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Agilent autosampler, by Agilent Technologies (1 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
Zorbax sb c18, by Agilent Technologies (1 mentions)
3 protocols using 1260 infinity quaternary hplc system
1
HPLC and LC-MS Analysis of Carotenoids and Apocarotenoids
2
HPLC Quantification of VRP in Brain
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The concentration of VRP in the brain tissue was measured by HPLC. The brain tissue samples were homogenized with a fourfold excess volume of distilled water. To 1.5 mL of the homogenate, 1 mL methanol was added. The samples were shaken for 5 min and centrifuged at 8000 rpm for 15 min. The organic layer was separated and filtered using a 0.45 μm membrane filter (Millipore, Ireland) prior to HPLC acquisition.
The HPLC system was consisted of a liquid chromatograph [Agilent™ 1260 Infinity Quaternary HPLC System equipped with Agilent High Performance ALS (G1367E), Agilent Quaternary Pump (G1311B/C), thermostat Agilent autosampler with reliable injections from 0.1 to 100 μL (G1329B), and Agilent fluorescence detector (G1321C) operated at 231 nm (excitation) and 318 nm (emission)]. The column used for HPLC was ZORBAX® Eclipse XDB C18 column (100 mm × 3 mm with 3.5 μm particle size) from Agilent. The mobile phase was acetonitrile-25 mM phosphate buffer (pH 4.0), methanol and acetonitrile (25:25:50, vol/vol). The flow rate of the mobile phase was 1.0 mL/min. The limit for quantification was 80 ng/mL in brain tissue.
The HPLC system was consisted of a liquid chromatograph [Agilent™ 1260 Infinity Quaternary HPLC System equipped with Agilent High Performance ALS (G1367E), Agilent Quaternary Pump (G1311B/C), thermostat Agilent autosampler with reliable injections from 0.1 to 100 μL (G1329B), and Agilent fluorescence detector (G1321C) operated at 231 nm (excitation) and 318 nm (emission)]. The column used for HPLC was ZORBAX® Eclipse XDB C18 column (100 mm × 3 mm with 3.5 μm particle size) from Agilent. The mobile phase was acetonitrile-25 mM phosphate buffer (pH 4.0), methanol and acetonitrile (25:25:50, vol/vol). The flow rate of the mobile phase was 1.0 mL/min. The limit for quantification was 80 ng/mL in brain tissue.
3
Analytical and Preparative HPLC Protocols
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NMR spectra were acquired in DMSO-d6 on a Bruker Avance II DRX-600K 600 MHz spectrometer at 25 °C. NMR spectra were processed using Topspin 3. Analytical HPLC was performed on a gradient Agilent 1260 Infinity quaternary HPLC system.
The column was an Agilent Zorbax SB-C18 (2.1 × 50 mm, 1.8 µm) eluted with a 0.6 ml min -1 gradient of 10-100% MeCN/H2O (0.01% TFA) over 8.33 min. Preparative HPLC was performed on a gradient Shimadzu HPLC system comprising of two LC-8 preparative liquid pumps with static mixer, SPD-M10AVP diode array detector and SCL-10AVP system controller with standard Rheodyne injection port. The columns used in the purification of the metabolites were selected from either a Vydac C18 column (50 × 100 mm, 5 m; Grace Discovery), a Zorbax SB-C18 column (50 × 150 mm, 5 m; Agilent) or an Alltima C18 (22 × 250 mm, 5 m, Grace Discovery) isocratically with MeCN/H2O mixtures containing 0.01% TFA modifier.
The column was an Agilent Zorbax SB-C18 (2.1 × 50 mm, 1.8 µm) eluted with a 0.6 ml min -1 gradient of 10-100% MeCN/H2O (0.01% TFA) over 8.33 min. Preparative HPLC was performed on a gradient Shimadzu HPLC system comprising of two LC-8 preparative liquid pumps with static mixer, SPD-M10AVP diode array detector and SCL-10AVP system controller with standard Rheodyne injection port. The columns used in the purification of the metabolites were selected from either a Vydac C18 column (50 × 100 mm, 5 m; Grace Discovery), a Zorbax SB-C18 column (50 × 150 mm, 5 m; Agilent) or an Alltima C18 (22 × 250 mm, 5 m, Grace Discovery) isocratically with MeCN/H2O mixtures containing 0.01% TFA modifier.
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