Several low-molecular-weight phenolic compounds were weighed (1.5 mg) and dissolved in 1 mL of 1:1 (v/v) methanol/water and membrane-filtered (0.45 μm pore size) with an HPLC coupled to a diode array detector (Agilent ChemStation, 1200, USA), equipped with a low-pressure quaternary pump (model Agilent 1200) and autosampler (model Agilent 1260 Infinity Autosampler). The separation was made at 20°C using a Nova-Pak C18 column at 300 mm and the injection volume was 25 μL of sample. All the phenols were determined by a direct readout of the sample absorbance at 280 nm. The photodiode array detector was set from 210 to 360 nm. Two mobile phases were used: A, water/acetic acid (98:2 v/v); B, water/acetonitrile/acetic acid (78:20:2 v/v/v), with a flow rate of 1 mL min−1 from 0 to 55 min and 1.2 mL min−1 from 55 to 85 min as follows: 100–20% A from 0 to 55 min, 20–10% A from 55 to 57 min, and 10–0% A from 57 to 85 min. Each major peak in the HPLC chromatograms was characterized by both the retention time and the absorption spectrum (from 210 to 360 nm).
Agilent 1260 infinity autosampler
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1260 Infinity autosampler is a liquid chromatography instrument designed to automatically introduce samples into the analysis system. It features precise sample handling and injection capabilities to ensure consistent and reliable data.
Automatically generated - may contain errors
Lab products found in correlation
Reference standard, by Merck Group (1 mentions)
Agilent 1260 diode array detector vl, by Agilent Technologies (1 mentions)
Zorbax sb c18, by Agilent Technologies (1 mentions)
Agilent hplc system, by Agilent Technologies (1 mentions)
1260 infinity diode array detector, by Agilent Technologies (1 mentions)
1260 infinity quaternary pump, by Agilent Technologies (1 mentions)
3 protocols using agilent 1260 infinity autosampler
1
HPLC Analysis of Phenolic Compounds
2
Silymarin Components Analysis by HPLC
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An Agilent 1260 Infinity High Performance Liquid Chromatography (HPLC) system equipped with an Agilent 1260 Infinity pump (G1361A), Agilent 1260 diode array detector VL (G1315D), Agilent ZORBAX SB-C18 and Agilent 1260 Infinity autosampler (G2260A) were used to analyze the content of individual silymarin components according to the reported method by (AbouZid et al., 2016b ). The mobile phase was composed of methanol/0.1% formic acid (Phase A) and water/0.1% formic acid (Phase B); 0–5 min gradient 40–45% phase A; 5–10 min isocratic 45% phase A; 10–20 min gradient 45–50% phase A; 20–25 min isocratic 50% phase A. The flow rate was 1.5 ml/min, and ambient temperature was used. The auto sampler was adjusted to inject 10 µl and quantified at 280 nm. The method offered the following retention times (minutes) for the major silymarin compounds: silychristin A (Rt = 8.9), silydianin (Rt = 10.6), silybin A (Rt = 19.6), silybin B (Rt = 21.3), isosilybin A (Rt = 25.4), and isosilybin B (Rt = 26.6). Flavonolignans identification and quantification was obtained using reference standards (Sigma-Aldrich). Total flavonolignan content is the result of the sum of the single constituents derived from HPLC analysis.
3
Quantitative Analysis of Synovial and Plasma Metabolites
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The synovial washing fluid and plasma samples were analyzed using an Agilent HPLC system (Agilent Technologies, Palo Alto, CA, USA) equipped with an automated solvent delivery system, which has an integrated degasser (1260 Infinity Quaternary Pump, Agilent Technologies), an Agilent 1260 Infinity autosampler (Agilent Technologies) and a 1024-element diode array detector (1260 Infinity Diode Array Detector, Agilent Technologies). The system control and data acquisition were performed with Agilent ChemStation B.04.03 software (Agilent Technologies). The chromatographic parameters and the sample extraction procedure have been provided in the Supplementary materials.
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