An adequate amount of the above isolated compound was dissolved in methanol and water to test its solubility. The methanol solution was processed for flavonoid characterization. First, magnesite powder and HCl were added into the methanol solution to test whether the compound was a flavonoid. Then, the solution was further evaluated by the Molisch reaction to determine whether it contained sugar. Then, the purity of the isolated compound was determined by HPLC. The structure was further analyzed by UV, ESIMS (Thermo Finnigan LCQ FLEET, Riviera Beach, FL, USA), 1H-NMR, and 13C-NMR (Avance III HD 400 MHz Digital NMR Spectrometer, Bruker, Karlsruhe, Germany).
Avance 3 hd 400 mhz digital nmr spectrometer
Manufactured by Bruker
Sourced in Germany
The Avance III HD 400 MHz Digital NMR Spectrometer is a laboratory instrument designed for nuclear magnetic resonance (NMR) spectroscopy. It operates at a frequency of 400 MHz and features a digital architecture.
Automatically generated - may contain errors
Lab products found in correlation
Lcq fleet, by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Cremophor el, by Maclin Biochemical Technology (1 mentions)
Peg 400, by Maclin Biochemical Technology (1 mentions)
Imark 96 well plate reader, by Bio-Rad (1 mentions)
β glucuronidase, by Merck Group (1 mentions)
Tween 80, by Maclin Biochemical Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Circular dichroism, by Applied Photophysics (1 mentions)
Triple toftm 5600 system, by AB Sciex (1 mentions)
4 protocols using avance 3 hd 400 mhz digital nmr spectrometer
1
Flavonoid Characterization and Purity Analysis
2
Deuterium Exchange and NMR Characterization
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An amount of 30 mg PPF was weighed precisely and deuterium-exchanged by freeze drying three time from 99% D2O to obtain the sample solution. A 0.5 mL portion of the sample solution was transferred to a 5 mm tube. NMR spectra were recorded in 25 °C on a Bruker NMR spectrometer (Avance III HD 400 MHz Digital NMR Spectrometer, Bruker, Karlsruhe, Germany).
3
Paclitaxel-Loaded Glucose Conjugate Synthesis
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Paclitaxel, succinate anhydride, 2,3,4,6-Tetra-O-benzyl-d -glucose, succinic anhydride, 4-dimethylaminopyridine (DMAP), dicyclohexylcarbodiimide (DCC), palladium on 10% activated carbon (10% Pd/C), chlorotriethylsilane (TESCl), tetrabutylammonium fluoride (TBAF), dichloromethane (DCM), tetrahydrofuran (THF), methanol (MeOH), acetic acid (AcOH), n-hexane (Hex), and ethyl acetate (EA) were purchased from Energy Chemicals (Shanghai, China). Cremophor EL, Tween-80 and PEG-400 were purchased from Macklin (Shanghai, China).
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium romide (MTT), dimethyl sulfoxide (DMSO), and β-glucuronidase were purchased from Sigma Aldrich (St. Louis, MO, USA). Leibovitz’s medium (L-15 medium), fetal bovine serum (FBS), and Hank’s balanced salt solution (HBSS) were purchased from Gibco (New York, NY, USA), rabbit serum (RS) was purchased from BestBio (Shanghai, China).
NMR spectra were recorded on a Bruker AVANCE III HD 400MHz Digital NMR Spectrometer (Karlsruhe, Germany). ESI mass spectra were recorded on a Thermo Scientific TSQ QuantumTM Access MAX Mass Spectrometer (Waltham, MA, USA). Optical Density (OD) of MTT assay was measured with a Bio-rad iMark 96-well plate reader (Hercules, CA, USA).
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium romide (MTT), dimethyl sulfoxide (DMSO), and β-glucuronidase were purchased from Sigma Aldrich (St. Louis, MO, USA). Leibovitz’s medium (L-15 medium), fetal bovine serum (FBS), and Hank’s balanced salt solution (HBSS) were purchased from Gibco (New York, NY, USA), rabbit serum (RS) was purchased from BestBio (Shanghai, China).
NMR spectra were recorded on a Bruker AVANCE III HD 400MHz Digital NMR Spectrometer (Karlsruhe, Germany). ESI mass spectra were recorded on a Thermo Scientific TSQ QuantumTM Access MAX Mass Spectrometer (Waltham, MA, USA). Optical Density (OD) of MTT assay was measured with a Bio-rad iMark 96-well plate reader (Hercules, CA, USA).
4
Structural Elucidation of Novel Gorgonian Metabolites
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Optical rotation data were measured on a MCP 200 dig-ital polarimeter (Anton Paar, Austria). UV and Circular Dichroism spectra were measured on a chirascan Circular Dichroism (Applied Photophysics, UK). HR-ESI-MS data were performed on a Triple TOFTM 5600 ＋ system (AB SCIEX, USA). NMR spectra were recorded on a Bruker avance III HD 400 MHz digital NMR spectrometer (Bruker Optics, Karlsruhe, Germany) with TMS as an internal standard. Column chromatography (CC) was performed with 200～300 mesh silica gel (Qingdao Marine Chemical Factory, China). TLC was performed on silica gel GF254 (Qingdao Marine Chemical Factory, China), spots were visualized under Ultraviolet transmission reflectometer (Shanghai Jingke Industrial, Shanghai). The preparative HPLC was performed on a Semi-preparative HPLC (QuikSep, Beijing, China) with Kromasil 100-5 C 18 column (10 mm×250 mm, 5 μm, Sweden). Subergorgic acid and 2-deoxy-2β-hydroxysubergorgic acid were used to establish GNPS, and their structures were determined by NMR and MS. 2-Deoxy-2β-hydroxysubergorgic acid was also from gorgonian Leptogorgia rigida. [13] The strains in this work were cultured in eight liquid media, eleven bioconversion and four rice media. All of culture media compositions are available in the Supporting Information (Tables S1～S3).
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