The NMR spectra were obtained with a Bruker 500 spectrometer ((Bruker, Bremen, Germany)) operating at 500 MHz for 1H and 125 MHz for 13C, respectively. Chemical shifts were reported in parts per million on the δ scale with TMS as the internal standard. The optical rotations were measured on a JASCO P-1020 Optical Rotation Apparatus (Jasco, Tokyo, Japan). ESI-HRMS spectra were measured on an Agilent 1100 LC/MSD TOF mass spectrometer (Agilent, California, USA). The CD spectrum were obtained on a JASCO810 spectropolarimeter (Jasco, Tokyo, Japan). Compounds 1 –7 were purified by semi preparative HPLC (Waters, Milford, USA) using a Waters 600 liquid chromatograph with a Alltech 2000Esc ELSD detector (110 °C, flow rate of the condensed air, 3.1 mL min−1) and with a Phenomenex HPLC column (4 μm, 4.6 × 250 mm, Phenomenex Hydro-RP 80R).
Agilent 1100 lc msd tof mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1100 LC/MSD TOF mass spectrometer is a high-performance liquid chromatography (LC) and time-of-flight (TOF) mass spectrometry (MS) system. It combines liquid chromatography with accurate mass measurement capabilities to provide precise identification and quantification of chemical compounds.
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2 protocols using agilent 1100 lc msd tof mass spectrometer
1
Spectroscopic Characterization of Novel Compounds
2
Quantification of Plant Secondary Metabolites
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Fresh tissues were extracted with 50% (v/v) methanol at 65°C for 1 h. Samples were centrifuged at 16,000 g for 5 min and then samples were analyzed. For HPLC analysis, 10 μL of extract was loaded onto a Shim-pack XR-ODS column (3.0 X 75 mm, 2.2 μm) (Shimadzu) using a gradient from 5% acetonitrile in 0.1% formic acid to 25% acetonitrile in 0.1% formic acid at a flow rate of 1 mL min -1 . Sinapic acid, cinnamic acid, kaempferol and cyanidin were used as standards for the quantification of sinpoylmalate, cinnamoylmalate, kaempferol glycosides, and anthocyanin, respectively. Liquid chromatography/mass spectrometry (LC/MS) analysis was conducted as following a previous report (Bonawitz et al., 2012) . Briefly, 10 μL of extract was run on the same Shim-pack XR-ODS column described above with a flow rate of 0.5 mL min -1 at an increasing concentration of acetonitrile from 10% to 25% over 14 min and from 25% to 70% over 8 min in 0.1% formic acid. Metabolites were analyzed with an Agilent 1100 LC/MSD TOF mass spectrometer in negative electrospray ionization (ESI) mode.
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