Ecs 400 400 mhz spectrometer

The HPLC system consisted of a Jasco (Hachioji, Japan) Model PU-2080 pump, a Jasco Model UV-2075 detector, a Rheodyne (Cotati, CA, USA) manual injector, a Shimadzu (Kyoto, Japan) column oven Model CTO-10Avp, and a Shimadzu degasser Model DGU-14A. InertSustain C18 column (5 μm, 4.6 mm i.d. × 150 mm, GL Sciences, Tokyo, Japan) was used. A mobile phase consisted of 20% acetonitrile, 20% methanol and 30 mM phosphate buffer (pH 7.0). Elution was carried out at a flow rate of 1.0 mL/min at 40°C. Analytes were detected at 254 nm. Data acquisition and processing were conducted with a Chromato-PRO (Runtime Instrument, Kanagawa, Japan). For LC/MS analysis, an LC 7400 series (Hitachi) equipped with an Agilent 6140 quadrupole mass spectrometer was used. LC/MS separation was performed on a InertSustain C18 column (3 μm, 2.1 mm × 250 mm, GL Sciences) with a mobile phase consisting of acetonitrile/water/ formic acid (40/60/0.1, v/v/v) at a flow rate of 0.2 mL/min at 40°C. ESI conditions (positive ion mode) were as follows: drying gas temperature, 250°C; drying gas flow, 10 L/min; capillary voltage, 4 kV. 1 H nuclear magnetic resonance (NMR) spectra were recorded on a JEOL ECS400 (400 MHz) spectrometer in CD3OD. Chemical shifts have been reported in δ ppm units with reference to the internal standard tetramethylsilane (Si(CH3)4, 0.00 ppm).