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Lc 8a series hplc system

Manufactured by Shimadzu

The LC-8a series HPLC system by Shimadzu is a high-performance liquid chromatography instrument designed for analytical applications. It features a compact and modular design, accommodating a variety of detectors and components to address diverse analytical requirements.

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3 protocols using lc 8a series hplc system

1

Analytical HPLC and LC-MS Characterization

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Analytical HPLC was performed on a Shimadzu LC-10Avp series HPLC system consisting of an autosampler, high-pressure pumps, column oven and PDA. HPLC conditions: C18 column (Eurospher 100-5, 250 × 4.6 mm) and gradient elution (MeCN/0.1 % (v/v) TFA 0.5/99.5 in 30 min to MeCN/0.1 % (v/v) TFA 100/0, MeCN 100 % for 10 min), flow rate 1 mL min−1. Preparative HPLC was performed on a Shimadzu LC-8a series HPLC system with PDA. LC-MS measurements were performed using an Exactive Orbitrap High Performance Benchtop LC-MS with an electrospray ion source and an Accela HPLC system (Thermo Fisher Scientific, Bremen). HPLC conditions: C18 column (Betasil C18 3 µm 150 × 2.1 mm) and gradient elution (MeCN/0.1 % (v/v) HCOOH (H2O) 5/95 for 1 min, going up to 98/2 in 15 min, then 98/2 for another 3 min; flow rate 0.2 mL min−1). NMR spectra were recorded on a Bruker AVANCE III 600 MHz instrument equipped with a Bruker cryo platform. The residual solvent signals were used as an internal reference.
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2

Analytical and Preparative HPLC-MS Characterization

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Analytical HPLC was performed on a Shimadzu LC-10Avp series HPLC system (Shimadzu Deutschland GmbH, Germany) consisting of an autosampler, high pressure pumps, column oven and a diode array detector. HPLC conditions: C18 column (Eurospher 100-5 250 × 4.6 mm; Alltech, Germany) and gradient elution (acetonitrile (MeCN)/0.1% (v/v) trifluoroacetic acid (TFA) (H2O) 0.5/99.5 in 30 min to MeCN/0.1% (v/v) TFA 100/0, MeCN 100% (v/v) for 10 min), flow rate 1 ml min−1. All solvents used were of analytical or HPLC grade. liquid chromatography-mass spectrometry (LCMS) measurements were performed using an Exactive Orbit-rap High Performance Benchtop LC-MS Mass Spectrometer with an electrospray ion source (Thermo Fisher Scientific, Germany). HPLC conditions: C18 column (Betasil C18 3 μm 150 × 2.1 mm; Thermo Fisher Scientific, Germany) and gradient elution (MeCN/0.1% (v/v) HCOOH (H2O) 5/95 for 1 min, going up to 98/2 in 15 min, then 98/2 for another 3 min; flow rate 0.2 ml min−1; injection volume: 3 μl). Preparative HPLC was performed on a Shimadzu LC-8a series HPLC system with DAD. NMR spectra were recorded on a Bruker Avance DRX 600 instrument (Bruker BioSpin GmbH, Germany). Spectra were normalized to the residual solvent signals.
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3

Preparative HPLC Analysis of Chloroform Fraction

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Preparative HPLC analysis of the chloroform fraction (CF) was carried using Shimadzu (LC-8A) series HPLC system, DI diode array detector (SP-M10AVP), reverse phase C18 column (20 mm × 250 mm). Samples (2 mL, 30 mg/mL) were injected into the column repeatedly and the reference molecule, quercetin was used as a standard (1 mL, 1 mg/mL). An Isocratic elution method was employed using a mobile phase of methanol, acetic acid and water in the ratio 18:2:80. The flow rate was 10 mL/min and the absorption was measured at a wavelength of 280 and 320 nm. From the gradient elution, four major peaks were obtained at retention time 20.5, 30.1, 37.5 and 57.1 min and were collected, and labelled CHF1 through CHF4. The collected isolated fractions were dried by removing the solvent using rotavapour at 40 °C. The samples were kept in a vacuum desiccator for removal of traces of water and then stored at 4 °C until further analysis.
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