Jnm a500 spectrometer

Microsome preparations from baculovirus-infected insect cells coexpressing CYP2C9*1, *2, or *3 with NADPH-CYP oxidoreductase were purchased from Gentest (Woburn, MA, USA); NADP, glucose-6phosphate (G6P), and G6P dehydrogenase were obtained from Oriental Yeast (Tokyo, Japan); and TB, 4'-hydroxytolbutamide, and all other chemicals were obtained from Sigma-Aldrich (St. Louis, Mo, USA). The melting point was determined using a MP-500D (Yamato Scientific, Tokyo, Japan) micro melting point apparatus without correction. Mass spectra (electron impact ionization) were measured using an AX505HA spectrometer (JEOL, Tokyo, Japan). 1 H-NMR spectra were recorded using a JNM-A500 spectrometer (JEOL) in CDCl 3 using tetramethylsilane as an internal standard.

1 H and 13 C NMR spectra were recorded on a JEOL JNM-A500 spectrometer operating at 500 and 125 MHz, respectively. Chemical shift values and coupling constants are reported as δ (ppm) and J (Hz), respectively. HPLC-PDA-MS analysis was performed using a JASCO Gulliver HPLC system equipped an MD-910 detector and a Waters ZQ 4000 mass spectrometer. The HPLC instrument was equipped with a Cosmosil 5C18-MS column (4.6 mm × 150 mm), and an elution rate of 0.8 mL∕ min was used. HPLC solvents consisted of water containing 0.1% TFA (solvent A) and methanol containing 0.1% TFA (solvent B). The elution protocol for analytical HPLC began with 100% A, followed by a linear gradient to 100% B over 30 min and held at 100% B for 10 min. Mass analyses were performed in electrospray ionization-positive and -negative modes. High-resolution mass analysis was performed using a Thermo Fisher Scientific, Inc. Orbitrap Velos ETD mass spectrometer. UV-visible-NIR absorption spectra were obtained on a JASCO V-530DS spectrometer using a quartz cell (Hellma, 10-mm light path length) or a micro cell (JASCO, 1.0-mm light path length). Fluorescence spectroscopic studies were performed either in a fluorescence cell (Hellma, 10 × 10 mm) or in a triangular fluorescence cell (JASCO FSA-410) with a JASCO FP-6600 spectrofluorometer.