The optical rotations were measured using a Jasco P2000 polarimeter, UV spectra were determined using a Jasco V650 spectrophotometer (JASCO, Corporation, Tokyo, Japan). IR spectra were carried out on a Nicolet 5700 spectrophotometer with KBr disks (Thermo Electron Scientific Instruments Corp.). 1H NMR (500 MHz), 13C NMR (125 MHz), HSQC and HMBC spectra were run on a Mercury-500 with TMS as an internal standard (Varian, Palo Alto, CA, USA). HR-ESI-MS was performed on a 6520 Accurate-Mass Q-TOF LC/MS mass spectrometer. Sephadex LH-20 (Pharmacia, Uppsala, Sweden), silica gel (Qingdao Marine Chemical Factory, 200–300 mesh), and RP-18 (Merck, 40—60 μm) were used for CC and silica gel GF-254 (Qingdao Marine Chemical Factory, Qingdao, China) was used for TLC. The HPLC experiments were performed on a preparative YMC-Pack ODS-column (250 mm×20 mm, 10 μm, YMC, Kyoto, Japan) equipped with a Shimadzu SPD-6A UV spectrophotometric detector and an SPD-6AD pumping system (Shimadzu, Japan).
Pack ods column
Manufactured by YMC
Sourced in Japan
The YMC-Pack ODS column is a reversed-phase high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a variety of organic compounds. The column utilizes an octadecyl (ODS) stationary phase, which provides a high degree of selectivity and retention for a wide range of analytes.
Automatically generated - may contain errors
Lab products found in correlation
Rp 18, by Merck Group (1 mentions)
Spd 6a uv spectrophotometric detector, by Shimadzu (1 mentions)
Mercury 500, by Agilent Technologies (1 mentions)
Silica gel, by Qingdao Marine Chemical (1 mentions)
Gf254, by Qingdao Marine Chemical (1 mentions)
P 2000 polarimeter, by Jasco (1 mentions)
V 650 spectrophotometer, by Jasco (1 mentions)
G1329b autosampler, by Agilent Technologies (1 mentions)
G1315d dad, by Agilent Technologies (1 mentions)
G1311b quaternary pump, by Agilent Technologies (1 mentions)
Agilent chemstation software, by Agilent Technologies (1 mentions)
1260 infinity hplc system, by Agilent Technologies (1 mentions)
Lc 20a, by Shimadzu (1 mentions)
Amazon iontrap mass spectrometer, by Bruker (1 mentions)
Agilent 1260 hplc system, by Agilent Technologies (1 mentions)
Lc 8a, by Shimadzu (1 mentions)
Lc 10a, by Shimadzu (1 mentions)
Prominence lc 20a hplc instrument, by Shimadzu (1 mentions)
8 protocols using pack ods column
1
Spectroscopic Characterization of Compounds
2
HPLC Analysis of Herbal Extract
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The high-performance liquid chromatography (HPLC) system consisted of an Agilent infinity series 1260 liquid chromatography system with a G1311B quaternary pump, G1329B autosampler, G1316A column oven and G1315D DAD detector, connected to Agilent ChemStation software (Agilent, Waldbronn, Germany). The separation was conducted with a YMC-PACK ODS column (4.6 mm × 250 mm, 5 μm). The gradient profile was as follows: 0–5 min, initial mobile phase of 0.1% formic acid prepared in acetonitrile and 0.1% formic acid prepared in water (10:90); 5–45 min, linear gradient of 70:30; and 45–50 min, isocratic 70:30. The flow rate was 1.0 ml/min, and detection was at 254 nm. The sample concentration was 10 mg/min in MeOH, and the injection volume was 10 μl. The HPLC chromatogram of SCE is shown in Fig. 1B .
3
HPLC Analysis of Chickpea Saponins
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HPLC analyses were performed on an Agilent 1260 Infinity HPLC system equipped with a G1311C quaternary pump, a G1329B autosampler, a G1316A thermostatted column compartment, and a G4260B evaporative light scattering detector (ELSD) coupled with an analytical workstation (Agilent Technologies, Inc., Santa Clara, CA, USA). Chromatographic separation was achieved on a YMC-Pack ODS column (250 mm × 4.6 mm i.d., 5 μm, YMC Co., Ltd., Kyoto, Japan). Methanol–acetonitrile–water containing 2% (v/v) acetic acid (40:25:35, v/v/v) was used as the mobile phase. The flow rate was 1.0 mL·min−1, the injection volume was 5 μL, and the column temperature was maintained at 25 °C. For ELSD detection, the carrier gas was air, the evaporating tube and atomization tube temperatures were set at 110 °C and 80 °C, respectively, with gas flow rate of 1.4 L·min−1. The chromatographic peak of chickpeasaponin B1 was verified by comparing its retention time with the reference standard. The content was calculated from the working calibration curve constructed by plotting the logarithm of peak areas vs the logarithm of the amounts injected series reference standard solutions.
4
Chromatographic Analysis of KQR
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Chemical analysis of KQR was performed using a liquid chromatograph model Shimadzu LC-20A (Shimadzu Corporation, Japan). The separation column model is YMC-Pack ODS column (YMC, Japan), and the mobile phase is 0.1% phosphoric acid aqueous solution (A)-acetonitrile (B), 5-45% acetonitrile gradient system, elution time 0-60 minutes, and 100% B, 60-70 minutes. The flow rate was 1 mL/min, and the test solution injection volume was 20 μL.
5
HPLC-MS Analysis of 11 Standard Compounds
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The separation of all 11 standard compounds was performed on a YMC‐Pack ODS column (250 mm × 4.60 mm, 5 μm) from YMC, using a mobile phase comprising 20 mM ammonium formate and 0.6% (v/v) formic acid in water (A) and methanol (B). Liquid chromatography ‐ diode array detector ‐ electrospray ionization ‐ mass spectrometry (LC‐DAD‐ESI‐MS) experiments were performed on an Agilent 1260 HPLC system (Santa Clara, CA, USA), which was coupled to an amaZon iontrap mass spectrometer (Bruker, Bremen, Germany). The HPLC instrument was equipped with binary pump, autosampler, column oven and diode array detector.
Elution was performed by maintaining 2% B for the first 15 min, followed by an increase to 10% B in 8 min, to 15% B in 7 min, and to 98% B in further 5 min; this composition was kept for additional 5 min, resulting in a total runtime of 35 min. The column was re‐equilibrated for 15 min prior to the next analysis. The DAD was set to 310, 330, and 350 nm, the flow rate, injection volume, and column temperature were adjusted to 0.65 mL · min−1, 5 μL, and 20°C. MS‐spectra were recorded in positive‐ESI mode (capillary voltage 4.5 kV), with a drying gas temperature of 200°C, the nebulizer gas (nitrogen) set to 4.4 psi, and a nebulizer flow (nitrogen) of 6 L · min−1. The scanned mass range was set between m/z 100 and 1,200.
Elution was performed by maintaining 2% B for the first 15 min, followed by an increase to 10% B in 8 min, to 15% B in 7 min, and to 98% B in further 5 min; this composition was kept for additional 5 min, resulting in a total runtime of 35 min. The column was re‐equilibrated for 15 min prior to the next analysis. The DAD was set to 310, 330, and 350 nm, the flow rate, injection volume, and column temperature were adjusted to 0.65 mL · min−1, 5 μL, and 20°C. MS‐spectra were recorded in positive‐ESI mode (capillary voltage 4.5 kV), with a drying gas temperature of 200°C, the nebulizer gas (nitrogen) set to 4.4 psi, and a nebulizer flow (nitrogen) of 6 L · min−1. The scanned mass range was set between m/z 100 and 1,200.
6
Eriocitrin Isolation from Lemon Peel
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Eriocitrin was prepared from lemon peel using the modified method of Miyake et al51 . The lemon peel was extracted using deionized water. The extract was applied to an Amberlite XAD-16 column (Rohm and Haas, Philadelphia, PA, USA). The column was washed with water, and eluted with 40% ethanol. The eluate was concentrated under reduced pressure and crude lemon flavonoids were obtained. Eriocitrin was prepared from crude flavonoids using preparative HPLC (LC-8A; Shimadzu, Kyoto, Japan) using a YMC-Pack ODS column (50 × 250 mm; YMC, Kyoto, Japan). The purity was determined as >96% using HPLC (LC-10A; Shimadzu).
7
HPLC Analysis of Chromatographic Samples
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Chromatographic analysis was performed using a SHIMADZU Prominence LC‐20A HPLC instrument (Shimadzu Corporation) equipped with a YMC‐ODS Pack column (4.6 mm × 250 mm, YMC Co., Ltd.). The detection wavelength was set at 202 nm and the temperature of the column oven was 25°C. The mobile phase was consisted of water (A) and acetonitrile (B). A gradient elution was used as follows: 5 min 10% B; 10 min 15% B; 15 min 20% B; 35 min 28% B; 50 min 40% B; 60 min 60% B; 70 min 70% B; and 75 min 10% B. The flow rate was kept at 1 ml/min, and the injected volume was 10 μl.
8
HPLC analysis of chemical compounds
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HPLC analysis was performed, the SHIMADZU Prominence LC-20A HPLC instrument (Shimadzu Corporation) and a YMC-ODS Pack column (4.6 mm × 250 mm, YMC Co., Ltd.) was used. The parameters of the HPLC were conducted according to the methods of Xue Peng.16 (link) The detection wavelength was set at 202 nm and the column oven was maintained at 25 °C. Solvent A was water and solvent B was acetonitrile. The linear UPLC gradient elution programs were used as follows: 5 min 10% B; 10 min 15% B; 15 min 20% B; 35 min 28% B; 50 min 40% B; 60 min 60% B; 70 min 70% B and 75 min 10% B. The separation was operated at a flow rate was of 1 mL min−1 and the injection volume was 10 μL.
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