The standard solutions of quercetin (1 mM) and extract of Ginkgo Folium, spiked with 0.5 mM quercetin, were prepared. Liquid chromatography was performed on an Agilent 1200 series system (Agilent, Waldbronn, Germany), which was equipped with a degasser, a binary pump, an autosampler, a DAD, and a thermostated column compartment. The herbal extract was separated on a Grace Prevail C-18 column (5 μm id, 250 mm × 4.6 mm). The mobile phase was composed of 0.1% formic acid in acetonitrile (A) and 0.1% formic acid in water (B) using the following gradient program: 0–90 min, linear gradient 25.0–60.0% (A); 90–95 min, linear gradient 60.0–75.0% (A); 95–120 min, linear gradient 75.0–95.0% (A). A preequilibration period of 4 min was used between each run. The flow rate was 0.75 mL/min. The column temperature was 30°C. The injection volume was 100 μL. The UV detector wavelength was set to 330 nm.
C18 column
Manufactured by Grace Bio-Labs
Sourced in United States
The C18 column is a chromatographic column used for separating and purifying a wide range of organic compounds. It contains a stationary phase made of silica particles coated with a C18 alkyl chain. This column is commonly used in high-performance liquid chromatography (HPLC) and other liquid chromatography techniques.
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Lab products found in correlation
Agilent 1200 series system, by Agilent Technologies (1 mentions)
Mini protean 2 electrophoresis cell, by Bio-Rad (1 mentions)
Beckman system gold, by Beckman Coulter (1 mentions)
C4 column, by Grace Bio-Labs (1 mentions)
C18 column, by Dikma Technologies (1 mentions)
Sb c18 column, by Agilent Technologies (1 mentions)
6 protocols using c18 column
1
Quantitative Analysis of Quercetin in Ginkgo Extracts
2
Antibiofilm Compounds Isolation via RP-HPLC
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In order to identify the antibiofilm components of ES products, SPE eluate was fractionated using a reverse phase (RP)-HPLC chromatography (Beckman System Gold, USA) coupled with a C18 column (250 mm × 4.6 mm; 5 μm) (Grace, USA) at a flow rate 0.3 mL/min, by using a gradient from 0 to 90% (v/v) acetonitrile (containing 0.1% (v/v) trifluoroacetic acid), during 70 min. The fractions were lyophilized and dissolved in 100 μL of TSB and tested for antibiofilm activity against S. aureus. The active fraction was then fractionated using a C4 column (250 mm × 4.6 mm; 5 μm) (Grace, USA) and tested under the same conditions. The purity of active fractions was checked by electrophoresis on 16.5% Tricine-SDS-PAGE gel using a Mini-Protean II electrophoresis cell (Bio-Rad, CA, USA). Protein bands were detected after staining with Coomassie Brilliant Blue R-250.
3
Optimization of C18 Column Conditions
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We tested the method with different columns, including Waters Symmetry C18 column, Xbridge C18 column, Cosmosil C18 column, Dikma C18 column, and Grace Alltima C18 column, all with dimensions 4.6 × 250 mm, 5 μm, under different temperatures, including 25°C, 30°C, and 35°C.
The following conditions were set: solvent A: acetonitrile, solvent B: 0.1% (v/v), 0.05% (v/v), and 0.2% (v/v) phosphoric acid; injection volume: 10 µL; flow rate: 1 mL/min; detector: photodiode array detector (PDA); mobile phase: 0–12 min, 75%–65% solvent B; 12–50 min, 65%–40% solvent B; 50–60 min, 40%–20% solvent B; 60–65 min, 20%–10% solvent B.
The following conditions were set: solvent A: acetonitrile, solvent B: 0.1% (v/v), 0.05% (v/v), and 0.2% (v/v) phosphoric acid; injection volume: 10 µL; flow rate: 1 mL/min; detector: photodiode array detector (PDA); mobile phase: 0–12 min, 75%–65% solvent B; 12–50 min, 65%–40% solvent B; 50–60 min, 40%–20% solvent B; 60–65 min, 20%–10% solvent B.
4
Radiosynthesis of [18F]18a via Kryptofix Method
5
HPLC Separation Using a C18 Column
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A Grace Prevail C18 column (5 μm, 10 mm × 250 mm i.d.) was used for separation by the method described in Table 1 at a flow rate of 2.0 mL/min. The column eluent was monitored at a wavelength of 254 nm. The injection volume was 50 μL.
6
LC-ESI and ESI-MS Mass Spectrometry Analysis
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Mass spectrometry analyses were carried out on a LC-ESI and ESI-MS n mass spectrometer as already described (Parrot et al., 2013) (link). A Prevail C18 column (5 μ, 250 × 4.6 mm, GRACE ® ) kept at 30 °C was used. For HPLC a gradient system was applied: A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile). The following gradient was applied at a flow rate of 1 mL/min in the HPLC system: initial, 99% A; 0.01-5 min, 90% A linear; 5-7 min, 90% A linear; 7-15 min 75% A linear; 15-17 min 75% A linear; 17-25 min 50% A linear; 25-27 min 50% A linear; 27-40 min 0% A linear; followed by washing and reconditioning of the column. A split to 0.2 mL/min was applied before the mass spectrometry system and 20 μg were injected. The MS n spectra were recorded during the HPLC run using the following conditions: MS/MS analysis with starting collision induced dissociation energy of 35 eV and data analyses were performed using Xcalibur v1.0.
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