Microtof qiii mass spectrometer

Extracts were also analyzed on an Agilent HPLC 1260 series system equipped with a Bruker microTOF QIII mass spectrometer using an Agilent Eclipse XDB C18 column (5 μm, 4.6 × 150 mm). Separation was performed at a flow rate of 0.5 mL/min with a 40 min linear gradient from 5 to 100% ACN in H₂O, both containing 0.1% (v/v) HCOOH. The column was then washed with 100% ACN for 5 min and equilibrated for 5 min with 5% ACN in H₂O. The parameters of the spectrometer were set as the following: electrospray positive ion mode for ionization, capillary voltage with 4.5 kV, collision energy with 8.0 eV. Sodium formate was used in each run for mass calibration. The masses were scanned in the range of m/z 100–1500. Data were evaluated with the Compass DataAnalysis 4.2 software (Bruker Daltonik, Bremen, Germany).

UV spectra were detected on an Alltech UVIS-200 detector. IR spectra were determined on a Thermo Nicolet Nexus Is50 FT-IR spectrometer. The NMR spectra were acquired on a 400, 500, or 600 MHz Bruker FTNMR spectrometer. Chemical shifts are referenced to the solvent peaks at δ_H 2.50 and δ_C 39.52 for DMSO-d₆. Mass spectra were obtained from a Bruker APEX IV 70 eV FT-MS spectrometer. The chromatographic (CC) substrates included silica gel (100−200 and 200−300 mesh) and HF254 silica gel for thin-layer chromatography (TLC) (Qingdao Marine Chemistry Co., Ltd., Qingdao, China). High-performance liquid chromatography (HPLC) was performed with an Alltech 426 pump equipped with an Alltech UVIS-200 detector (210 nm) and using a semipreparative reversed-phase column (YMC-packed, C₁₈, 5 μM, 10 × 250 mm). UPLC was performed on an Agilent UPLC series 1200 (Agilent Technologies, Santa Clara, CA, USA) equipped with an Agilent Eclipse XDB-C18 column (5 μm, 4.6 × 150 mm). LC-MS analysis was carried out on an Agilent HPLC 1260 series system equipped with a Bruker microTOF QIII mass spectrometer by using an Agilent Eclipse XDB C18 column (5 μm, 4.6 × 150 mm) or Waters UPLC-MS system. The chemicals used in this study were obtained from Beijing Tongguang Fine Chemicals Company of the highest available purity.