General method. Synthesis reagents and solvents were obtained from commercial suppliers and used without further purification. Characterizations of compounds are provided in the Supporting Information. 1H and 13C NMR spectra were recorded on Bruker (400 MHz) instruments, using dimethyl sulfoxide (DMSO‑d6) as solvents. Chemical shifts are given in parts per million (ppm) downfield from tetramethylsilane (δ) as the internal standard in deuterated solvent and coupling constants (J) are in Hertz (Hz). Data are reported as follows: chemical shift, integration, multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublet, ddd = doublet of doublet of doublet, dt = doublet of triplet, m = multiplet, bs = broad signal), and coupling constants. High-resolution mass spectra were recorded on a Bruker Apex IV FTMS mass spectrometer using electrospray ionization (ESI). All compounds tested in biological assays were >95% pure.
Apex 4 ftms mass spectrometer
Manufactured by Bruker
Sourced in Germany
The Apex IV FTMS mass spectrometer is a high-performance Fourier Transform Mass Spectrometer (FTMS) designed for advanced analytical applications. It offers high mass accuracy, resolution, and sensitivity for the detection and analysis of a wide range of molecular species.
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3 protocols using apex 4 ftms mass spectrometer
1
General Method for Compound Characterization
2
Quantitative Analysis of Caffeic Acid Derivatives
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Separation and collection of CA were performed on a Gilson analytical and semi-preparative high performance liquid chromatography (HPLC) which comprised a 322 Pump and a 152 UV/Vis detector (Gilson Science & Technology, Beijing, PR China). Separation and collection of caffeoylquinic acid compounds (CACs) were performed on a HD 21C-B UV detector and a HDC-01 signal acquisition unit from Shanghai Jihui Scientific Analytical Instruments Co., Ltd (Shanghai, PR China). The HPLC experiments were conducted on a HP 1100 series HPLC (Agilent Technologies, USA). A Shimadzu UV-2550 Ultraviolet and visible spectrophotometer was used for the batch adsorption experiments (Kyoto, Japan). High-resolution mass spectra (HRMS) were detected by a Bruker Apex IV FTMS mass spectrometer (ESI) (Bruker Daltonics, Germany).
The optical density of cell experiment was measured by a SpectraMax M2e Microplate Reader (Molecular Devices, Menlo Park, USA).
The optical density of cell experiment was measured by a SpectraMax M2e Microplate Reader (Molecular Devices, Menlo Park, USA).
3
Characterization of TNFR1-ECD binding compounds
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HEK293T cells were received as a gift from Professor Jincai Luo (Peking University, China). The extracellular domain of the TNF receptor 1 (TNFR1-ECD) was purchased from R&D Systems. The selected compounds were purchased from the SPECS database with purity higher than 90% and for most compounds >95% (confirmed by the supplier, using NMR or LC-MS data available through the website). Other biochemistry reagents were from Sigma Aldrich unless indicated otherwise. The organic reagents and solvents were commercially available and purified according to conventional methods. All reactions were monitored by thin layer chromatography (TLC), using silica gel 60 F-254 aluminum sheets and UV light (254 and 366 nm) for detection. All title compounds gave satisfactory 1H NMR, 13C NMR, and mass spectrometry analyses. The 1H NMR and 13C NMR spectra were measured on a Bruker-400 M spectrometer using TMS as internal standard. High resolution mass spectra were recorded on a Bruker Apex IV FTMS mass spectrometer using ESI (electrospray ionization).
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