The 10% fraction was used to purify individual C3G and the 50% fraction was used to purify the three flavonols by Prep-HPLC (2535 pump, 2998 detector, 2707 auto-sampler, Waters, Milford, MA, USA) with a SunFireTM prep C18 OBDTM (5 μm, 19 mm × 250 mm) column, respectively. The mobile phase solvents were the same as that of HPLC. The C3G gradient program was as follows: 0–10 min, 10% B; 10–30 min, 10%–30% B; 30–50 min, 30%–45% B; 50–60 min, 45%–60% B. The flavonols gradient program was as follows: 0–35 min, 20%–38% B; 35–60 min, 38%–48% B, 60–80 min, and 48%–100% B. The flow-rate was 5.0 mL/min, the injection volume was 300 μL and the concentration was 100 mg/mL. The effluent from the outlet of the column was monitored at 280 nm and 350 nm. The peak fractions were collected according to the chromatogram.
2535 pump
Manufactured by Waters Corporation
Sourced in United States
The 2535 pump is a high-performance liquid chromatography (HPLC) pump manufactured by Waters Corporation. It is designed to deliver precise and consistent flow rates for a variety of HPLC applications. The 2535 pump features a dual-piston design and advanced pulse dampening technology to ensure smooth and stable solvent delivery.
Automatically generated - may contain errors
Lab products found in correlation
Silica gel, by Qingdao Haiyang Chemical (3 mentions)
Gf254, by Qingdao Haiyang Chemical (3 mentions)
2707 autosampler, by Waters Corporation (1 mentions)
1525 separation module, by Waters Corporation (1 mentions)
Av 600 mhz spectrometer, by Bruker (1 mentions)
Autopol 6, by Rudolph Research Analytical (1 mentions)
Proc18 column, by Waters Corporation (1 mentions)
2414 refractive index detector, by Waters Corporation (1 mentions)
Ascend 600 nmr spectrometer, by Bruker (1 mentions)
Packed c18, by YMC (1 mentions)
Genesys 10s uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Semi preparative hplc, by Waters Corporation (1 mentions)
Maxis esi qtof mass spectrometer, by Bruker (1 mentions)
Agilent xcalibur atlas gemini ultra diffractometer, by Agilent Technologies (1 mentions)
J 810 spectropolarimeter, by Jasco (1 mentions)
Av 400 spectrometer, by Bruker (1 mentions)
4 protocols using 2535 pump
1
Purification of Anthocyanins and Flavonols
2
Analytical Techniques for Natural Products
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Optical rotation measurements
were determined using an Autopol VI, serial no. 91003, manufactured
by Rudolph Research Analytical Hackettstown, NJ, USA. IR and UV spectra
were obtained on a Thermo Fisher Nicolet iS50 spectrometer and a Persee
TU-1950-YV-VIS spectrophotometer, respectively. ECD spectra were recorded
on a Jasco-J-810 spectropolarimeter. NMR spectroscopic data were acquired
on a Bruker AV-600 MHz spectrometer in methanol-d4 (δH 3.31; δC 49.0).
HRESIMS was obtained in positive ion mode on a Waters Xevo G2-XS/APGC
spectrometer. Preparative MPLC was used in a Interchim Puriflash 450
instrument. Column chromatography (CC) was carried out using silica
gel (200–300 mesh; Yantai) and ODS C18 (15 μm,
Santai Technologies, Inc.). Analytical thin-layer chromatography was
performed on a silica gel FSGF254 plate. A YMC-Pack Pro
C18 column (250 × 10 mm, 5 μm) was used for
reversed-phase HPLC using a Waters 1525 separation module equipped
with a Waters 2535 pump and a 2998 photodiode array detector.
were determined using an Autopol VI, serial no. 91003, manufactured
by Rudolph Research Analytical Hackettstown, NJ, USA. IR and UV spectra
were obtained on a Thermo Fisher Nicolet iS50 spectrometer and a Persee
TU-1950-YV-VIS spectrophotometer, respectively. ECD spectra were recorded
on a Jasco-J-810 spectropolarimeter. NMR spectroscopic data were acquired
on a Bruker AV-600 MHz spectrometer in methanol-d4 (δH 3.31; δC 49.0).
HRESIMS was obtained in positive ion mode on a Waters Xevo G2-XS/APGC
spectrometer. Preparative MPLC was used in a Interchim Puriflash 450
instrument. Column chromatography (CC) was carried out using silica
gel (200–300 mesh; Yantai) and ODS C18 (15 μm,
Santai Technologies, Inc.). Analytical thin-layer chromatography was
performed on a silica gel FSGF254 plate. A YMC-Pack Pro
C18 column (250 × 10 mm, 5 μm) was used for
reversed-phase HPLC using a Waters 1525 separation module equipped
with a Waters 2535 pump and a 2998 photodiode array detector.
3
Isolation and Structural Elucidation of Natural Compounds
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HRESIMS were determined by Q-TOF-MS system (SCIEX, Framingham, MA, USA). The NMR spectra were recorded on Bruker Ascend 600 NMR spectrometer (Bruker, Bremen, Germany), using TMS as an internal standard. HPLC purifications were performed on an analytical reversed-phase column (YMC-packed C18, 250 mm × 10 mm, 5 μm) (YMC, Tokyo, Japan) using a Waters 2535 Pump and detected with a Waters 2414 refractive index detector. Column chromatography was performed with silica gel (100–200, 200–300 mesh, Qingdao Haiyang Chemical Co., Ltd., Qingdao, China), ODS-A gel (S-20 μm, Beijing Jinouya Chemical Co., Ltd., Beijing, China), TLC was carried out on silica gel GF254 (Qingdao Haiyang Chemical Co., Ltd., Qingdao, China). Methanol was purchased from Merck Company. Ethanol, petroleum ether, ethyl acetate and dichloromethane were purchased from Tianjin Fuyu chemical Co. Ltd. (Tianjin, China)
4
Comprehensive Analytical Characterization
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Optical rotations were recorded on a MCP500 modular circular polarimeter (Anton Paar Opto Tec GmbH, Seelze, Germany). UV spectra were obtained on a GENESYS 10S UV-Vis spectrophotometer (Thermo Fisher Scientific, Shanghai, China). HRESIMS were measured on a Bruker maXis ESI-QTOF mass spectrometer (Bruker Daltonics, Bremen, Germany). NMR spectra were recorded on a Bruker AV-400 spectrometer with TMS as the internal standard. Single-crystal X-ray diffraction analyses were made on an Agilent Xcalibur Atlas Gemini Ultra-diffractometer (Agilent Technologies, Santa Clara, CA, USA) with mirror monochromated CuKα radiation (λ = 1.54178 Å) at 150 K. Semi-preparative HPLC (Waters Corporation, Milford, MA, USA) was performed on a Waters 2535 pump equipped with a waters 2998 photodiode array detector and YMC C18 reverse-phased columns (250 mm × 10 mm i.d., 5 μm). For column chromatography, silica gel (100–200 mesh) (Qingdao Mar. Chem. Ind. Co. Ltd., Qingdao, China) and C18 reverse-phased silica gel (ODS-A-HG 12 nm, 50 µm, YMC Co. Ltd., Kyoto, Japan) were used. ECD spectra were measured on a Jasco J-810 spectropolarimeter (JASCO Corporation, Tokyo, Japan) in MeCN.
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