Inertsil ods 3 octadecylsilane column

An HPLC system including an SPD-M10Avp UV/Vis photodiode-array detector (Shimadzu, Kyoto, Japan) with an Inertsil ODS-3 octadecylsilane column of size 4.6 × 250 mm and particle size of 5 μm (GL Sciences, Tokyo, Japan) was used. The eluent was 20 mM ammonium acetate (pH 7.0) containing methanol. The methanol concentration was increased from 0 to 50% over 30 min in linear gradient mode. The column temperature was 40°C and the flow rate was 1 mL/min. The RP-HPLC chromatogram was detected at 200–500 nm. ESI-TOF-MS and HR-ESI-TOF-MS measurements were performed on a MicrOTOF spectrometer (Bruker, Bremen, Germany) in negative mode. The sample isolated by RP-HPLC was directly infused into the MS system by a syringe pump without a column. The NMR spectra were obtained on a JNM-ECA500 spectrometer (JEOL, Tokyo, Japan).

The HPLC system consisted of LC-10ADvp pumps and an SPD-M10Avp UV-Vis photodiode-array detector (Shimadzu, Kyoto, Japan). For the RP-HPLC, an Inertsil ODS-3 octadecylsilane column of size 4.6 × 250 mm and particle size of 5 μm (GL Sciences, Tokyo, Japan) was used. The eluent was 20 mM ammonium acetate (pH 7.0) containing acetonitrile. For AcdGuo, the acetonitrile concentration was increased from 0 to 30% over 45 min in linear gradient mode. For AcdAdo, the acetonitrile concentration was increased from 0 to 37.5% over 45 min in linear gradient mode. The column temperature was 40 °C and flow rate was 1 mL/min. The RP-HPLC chromatogram was detected at 200–500 nm. ESI-TOF/MS measurements were performed on a MicrOTOF spectrometer (Bruker, Bremen, Germany) in negative mode. The sample isolated by RP-HPLC was directly infused into the MS system by a syringe pump without a column.