Extracts isolation was conducted by binary HPLC (high-performance liquid chromatography) pump (HPLC WATERS™ 600, Milford, MA, USA) coupled with a WATERS 996 photodiode array (PDA) UV/Vis detector and the reversed-phase semi-prep HPLC isolation condition (Phenomenex Luna 5µ C18 column, 250 mm × 10 mm, 5 µm, eluting with 40% CH3CN at flow rate 2.0 mL/min). ESI (electrospray ionization) low-resolution LC-MS data were obtained with an Agilent Technologies 6120 quadrupole mass system (Agilent Technologies, Santa Clara, CA, USA) coupled with an Agilent Technologies 1260 series HPLC with a reversed-phase Phenomenex luna C18 column (4.6 mm × 100 mm, 5 µm) at a low flow rate of 1.0 mL/min. NMR spectra were acquired by Bruker Avance 300 MHz and 150 MHz spectrometers (Bruker Biospin Group, Karlsruhe, Germany) using methanol-d4 as a solvent, which was purchased from Cambridge Isotope Laboratories, Inc. (Tewksbury, MA, USA). For extract fractionation, first grade solvents were acquired from Dae-Jung chemicals & Metals Co. Ltd. For LC-MS and HPLC analyses, HPLC-grade solvents were provided from J.T.Baker. and Dae-Jung chemicals & Metals Co. Ltd. (Sheung-Si, Korea).
Luna 5 c18 column
Manufactured by Phenomenex
Sourced in United States
The Luna 5μ C18 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. It features a 5-micron particle size and a C18 stationary phase, which provides efficient and reproducible chromatographic separations.
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Lab products found in correlation
1260 series hplc, by Phenomenex (1 mentions)
Methanol d4, by Cambridge Isotopes (1 mentions)
Luna c18 column, by Phenomenex (1 mentions)
500 spectrometer, by Agilent Technologies (1 mentions)
Cary eclipse, by Agilent Technologies (1 mentions)
Zetasizer nano s, by Malvern Panalytical (1 mentions)
Quemesa, by EMSIS (1 mentions)
Em 208, by Philips (1 mentions)
Radius 2, by EMSIS (1 mentions)
8 protocols using luna 5 c18 column
1
Isolation and Identification of Bioactive Compounds
2
Synthesis and Purity Analysis of Compounds
3
HPLC-MS Metabolomic Analysis of Fungal Extracts
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The culture filtrates and mycelia extracts (CEs and MEs) were analyzed by an Agilent (Cernusco sul Naviglio, Milan, Italy) HPLC-MS ESI-TOF 1260/6230DA instrument operating in positive ionization mode. The source temperature was kept at 120 °C and the desolvating gas at 250 °C. The instruments were interfaced to a Phenomenex Luna 5µ C18 column (150 × 4.6 mm, 5 µm). An acetonitrile-water (0.1% formic acid) gradient was used starting from 30% acetonitrile, increasing linearly to 60% in 30 min, 60% for 10 min and finally rebalanced to the initial rate for 5 min. Methanol was used to dissolve the samples. HPLC flow rate was 500 µL min−1.
Pure compounds employed as analytical standards for the qualitative analysis were obtained as previously reported [25 (link),27 (link)].
Pure compounds employed as analytical standards for the qualitative analysis were obtained as previously reported [25 (link),27 (link)].
4
HPLC Analysis of Phenolic Compounds
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HPLC analyses were performed using a Shimadzu prominence series HPLC system (Phenomenex, Torrance, CA) equipped with a UV-Vis detector. A reversed phase Phenomenex Luna 5 C18 column (250 × 4.6 mm particle size) (Torrance, CA) was used to carry out the separation. The mobile phase was made up of two solvents: Solvent A was water, phosphoric acid, methanol (HPLC grade, Merck, Germany) in 89.7:0.3:10 (v/v/v) respectively and Solvent B was acetonitrile, all running in a gradient setting. After filtering the extracts with an 0.2-micron syringe filter (Acrodisc, India), they were stored at -20 ºC until use. Each sample was run for 20 min at a flow rate of 1 ml/min. The column was kept at a constant 25 ºC (± 2) temperature. Phenolic components were identified and quantified by comparing retention times and spectra with those of commercially available standard compounds.
5
HPLC Quantification of JS-K and TMZ in Liposomes
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The content of JS‐K and TMZ in S1P/JS‐K/TMZ/Lipo was determined using high performance liquid chromatography (HPLC) method. Briefly, freshly prepared liposomes were mixed with methanol solution (1:9 v/v) and then vortexed for 5 min to fully destroy the liposome structure. The JS‐K and TMZ were released from the liposomes. After filtration via a 200 nm membrane, the solution (20 µL) was measured by an HPLC system with a Phenomenex Luna 5 C18 column (Torrance, CA). The mobile phase was methanol‐acetic acid 0.5% (30:70, v/v) at a flow rate of 1 mL min−1. The concentration of the drug encapsulated inside the liposomes was calculated based on a JS‐K or TMZ standard concentration curve.
6
Bile Acids Binding Capacity of Dry-Cured Ham
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The bile acids binding capacity of the dry-cured ham extract was analyzed by reverse phase HPLC using a Luna5 C18 column (250 × 4.60 mm) from Phenomenex [18 (link)]. Glycocholic acid was used as standard to control the binding capacity of the extract.
7
Benzimidazole Metabolite Quantification
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Mice were given a bolus injection of 50 or 150 mg/kg drug prepared in DMSO or DNTC formulation before collection of plasma samples 8 h post injection. Benzimidazoles and their active metabolites (both sulfones and sulfoxides) in the plasma were extracted by liquid-liquid partition using potassium carbonate, DMSO, sodium metabisulfite and ethyl acetate, dried and analyzed by high performance liquid chromatography (HPLC) on a 150 x 4.6 mm Phenomenex Luna C18 5-µ column. Fenbendazole and its metabolites were detected using UV light (290 nm), while albendazole and its metabolites were detected using fluorescence (excitation at 290 nm; emission at 330 nm).
8
Biophysical Characterization of Nanomaterials
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NMR spectra (500 MHz) were recorded on a Varian 500 spectrometer (Palo Alto, CA, USA). HPLC analyses were run on an Agilent series 1100 liquid chromatograph (Santa Clara, CA, USA), with a Phenomenex (Torrance, CA, USA), Luna C18-5µ column with a column guard and a 20 µL loop. Fluorimetric titrations were performed on a CARY Eclipse (Varian) spectrometer with orthogonal geometry with a square cuvette of 0.5 cm optical path. TEM images were recorded with a Camera Olympus QUEMESA (Tokyo, Japan) and software RADIUS 2.0 (EMSIS) (Münster, Germany) on a TEM images Philips EM208 (Amsterdam, The Netherlands) at 100 kV using a 200-mesh copper grid with carbon film. Dynamic Laser Light Scattering was performed on a Zetasizer nano-S (Malvern Panalytical, Malvern, UK) instrument. Data analysis and regressions were carried out using Sigma Plot 13 (Sysdat Software Inc., San Jose, CA, USA).
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