Dried roots (500 mg) were extracted using ethanol: water (70:30), followed by sonication for half an hour in an ultrasonic bath. The filtered extracts were centrifuged and analyzed by high-performance liquid chromatography (HPLC)15 (link). The contents of glycyrrhizic acid and glabridin in the roots were determined using an Agilent liquid chromatography instrument, consisting of a 2695 separation module (Germany) equipped with a 100 μl loop and photodiode array detectors (PDA) using a C18 column (Knauer, 25 cm × 4.6 mm Eurospher 100–5) with an aqueous solution of 0.3% H3PO4 (solvent A) and with acetonitrile (solvent B) as the mobile phase. The flow rate was 1 mL/min with a linear solvent gradient of A–B as follows: 80% A for 10 min; reduced to 20% in 30 min and maintained for 5 min. The samples were measured at a wavelength of 230 nm for glabridin and 250 nm for glycyrrhizic acid. Each sample had a 20 μl injection volume for HPLC analysis. Millennium 32 (software) was used for data acquisition. The calibration curves for the standards were created by plotting the peak areas against the concentrations15 (link).
Liquid chromatography instrument
Manufactured by Agilent Technologies
Sourced in Germany
The Liquid Chromatography Instrument is a laboratory equipment used for the separation, identification, and quantification of components in a liquid mixture. It operates by passing the sample mixture through a column filled with a stationary phase, where the components are separated based on their different interactions with the stationary phase and the mobile phase.
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2 protocols using liquid chromatography instrument
1
Quantification of Glycyrrhizic Acid and Glabridin in Licorice Root Extracts
2
Polymer Characterization by Multitechnique Analysis
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NMR
measurements were performed at room
temperature using a Bruker AVANCE III-500 MHz spectrometer instrument
in CDCl3 solvent. Size exclusion chromatography was performed
on an Agilent liquid chromatography instrument equipped with two similar
PL gel columns connected in series and two detectors, namely, a refractive
index detector and a UV–Vis detector. DMF was used as a mobile
phase (containing 0.005 M LiBr) at a flow rate of 1 mL min–1 at 45 °C, and the polymer concentration was 2 mg mL–1. The instrument was calibrated with polystyrene standards. UV–Vis
absorption spectra were recorded on a Thermo Evolution 600 UV–visible
spectrophotometer. Fluorescence emission spectra were recorded using
a Horiba Jobin Yvon Fluoromax-4 spectrofluorometer. Dynamic light
scattering (DLS) measurements were carried out at 25 °C using
a Malvern Zetasizer Nano ZS instrument equipped with a 30 mW He–Ne
laser light source emitting vertically polarized light of 632.8 nm
wavelength (scattering angle 173°). Transmission electron microscopy
images were recorded on an FEI-Technai Twin instrument operated at
120 kV in the bright-field mode. The aqueous solution of the sample
(1 mg mL–1) was drop-cast on a carbon-coated copper
grid followed by staining with uranyl acetate (2%).
measurements were performed at room
temperature using a Bruker AVANCE III-500 MHz spectrometer instrument
in CDCl3 solvent. Size exclusion chromatography was performed
on an Agilent liquid chromatography instrument equipped with two similar
PL gel columns connected in series and two detectors, namely, a refractive
index detector and a UV–Vis detector. DMF was used as a mobile
phase (containing 0.005 M LiBr) at a flow rate of 1 mL min–1 at 45 °C, and the polymer concentration was 2 mg mL–1. The instrument was calibrated with polystyrene standards. UV–Vis
absorption spectra were recorded on a Thermo Evolution 600 UV–visible
spectrophotometer. Fluorescence emission spectra were recorded using
a Horiba Jobin Yvon Fluoromax-4 spectrofluorometer. Dynamic light
scattering (DLS) measurements were carried out at 25 °C using
a Malvern Zetasizer Nano ZS instrument equipped with a 30 mW He–Ne
laser light source emitting vertically polarized light of 632.8 nm
wavelength (scattering angle 173°). Transmission electron microscopy
images were recorded on an FEI-Technai Twin instrument operated at
120 kV in the bright-field mode. The aqueous solution of the sample
(1 mg mL–1) was drop-cast on a carbon-coated copper
grid followed by staining with uranyl acetate (2%).
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