The active compounds of the extracts and nanoparticles produced of G. lucidum were identified by Liquid Chromatography-Mass Spectrometry (LC–MS), based on the method proposed by Yang et al.34 (link). The using device in this work was an Agilent (Waldbronn, Germany) 1200 series chromatographic system coupled with an Agilent 6410 triple quadrupole tandem mass spectrometer. LC–MS data were analyzed using the software provided by Agilent Mass Hunter Workstation. An Agilent HT Zorbax SB-C18 column (5 µm particle size, 4.6 mm × 250 mm; Agilent Technologies, Santa Clara, CA) was carried out for the separation process. The mobile phase used in the gradient elution program consisted of acetonitrile and water containing 0.2% acetic acid(v/v) at a flow rate of 1 mL/min, and the injection volume was 5 μL. During 40 min, the volume of acetonitrile increased from 30 to 32% and reached 40% in 20 min. Then it was kept constant at this value for 5 min. The Electron Spray Ionization (ESI) was worked Positive ion mode with the voltage of 4 kV and a capillary temperature equal to 300 °C.
1200 series chromatographic system
Manufactured by Agilent Technologies
Sourced in Germany
The 1200 series chromatographic system is a high-performance liquid chromatography (HPLC) instrument manufactured by Agilent Technologies. It is designed to separate, identify, and quantify components within a sample mixture. The system includes a pump, an autosampler, a detector, and a data system to control the instrument and analyze the results.
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3 protocols using 1200 series chromatographic system
1
Identification of Active Compounds in Ganoderma lucidum
2
LC-MS/MS Analysis of Biomolecular Samples
3
HPLC and TEM Analysis of Nanomaterial
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High performance liquid chromatography (HPLC) was performed using Agilent Technologies 1200 series chromatographic system, which consisted of a vacuum degasser, a quaternary pump, a standard and preparative auto-sampler, a column compartment with a thermostat and a variable wavelength UV detector. The flow rate was set at 1.5 mL min -1 . Samples (10 μL) were chromatographed over a reverse phase column (Onyx Monolithic C18 column, 100 x 4.6 mm) fitted with a guard column maintained at 40 °C, and monitored for absorption at 227 nm wavelength. Samples were diluted in mobile phase (acetonitrile: water, 1: 1) and analysed using a standard curve (y = 36.79x -0.0857, r 2 = 0.998) with a concentration range of 0.1 -1.0 μg mL -1 . The particle size and particle size distribution were measured by Photon Correlation Spectroscopy (Malvern Zetasizer 3000HSA, Malvern Instruments, UK.) at 25˚C at a wavelength of 633 nm and the data analysed using the Contin method of analysis.
Measurements were performed in triplicate.
Transmission electron microscopy was performed using Philips/FEI CM120 Bio Twin (Philips, Netherlands). A drop of the formulation was dried on a copper TEM grid (300 mesh-Fomvar/ carbon coated) and stained with a drop of uranyl acetate (1% w/v, negative staining). Once dried, the samples were imaged and the representative images were photographed and documented.
Measurements were performed in triplicate.
Transmission electron microscopy was performed using Philips/FEI CM120 Bio Twin (Philips, Netherlands). A drop of the formulation was dried on a copper TEM grid (300 mesh-Fomvar/ carbon coated) and stained with a drop of uranyl acetate (1% w/v, negative staining). Once dried, the samples were imaged and the representative images were photographed and documented.
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