Jms t100 gcv

Optical rotations were obtained with a JASCO P-2200 polarimeter. NMR spectra were recorded in C₆D₆, CD₃OD and CDCl₃ using a JNM-EX 270 FT-NMR spectrometer (JEOL, ¹H NMR: 270 MHz) and AMX 500 Bruker system (¹H NMR: 500 MHz, ¹³C NMR: 126 MHz). Assignment of H and C was performed by obtaining ¹H NMR, ¹³C NMR (referenced for C₆D₆, CD₃OD and CDCl₃ at δ_H 7.16, 3.31 and 7.24, and δ_C 128.4, 49.2 and 77.2, respectively), COSY, HSQC, HMBC, and NOESY spectra. FD-MS analysis was performed on a JMS-T100GCV (JEOL) instrument. Chromatographic analysis was performed using an HPLC system (InertSustain, A_210max nm) equipped with a Shisheido Capcell park C18 column (4.6 × 250 nm, 5 µm, 2 mL/min, MeOH-H₂O, 80:20) and a Cadenza CK-C18 column (6 × 250 nm, 3 µm, 2 mL/min, MeOH-H₂O, 80:20). All moisture-sensitive reactions were performed under a nitrogen gas atmosphere. All chemicals used in the study were of analytical grade and purchased from Sigma–Aldrich, Tokyo, Japan, Kanto Chemical Co., Inc, Tokyo, Japan, and Cayman Chemical, Ann Arbor, MI, USA.

¹H-NMR spectra were recorded in CDCl₃ on a JEOL ECS-400 (400 MHz) spectrometer; tetramethylsilane was used as the internal reference. Electron ionization (EI) and electrospray ionization (ESI) mass spectrometry were performed using JEOL JMS-T100 GCv and JEOL JMS-T100 LP instruments, respectively. Elemental analyses were performed using MICRO CORDER JM10. UV-vis absorption spectra for ligand 1 and Eu(III) complex 2 were measured using a JASCO V-670 spectrophotometer. Emission spectrum, excitation spectrum, and emission lifetime for Eu(III) complex 2 were measured using a Horiba FluoroLog®3 spectrofluorometer. Emission spectrum and lifetime for Gd(III) complex 5 were measured using a FP-6300 spectrofluorometer with a nitrogen bath cryostat (Oxford Instruments, Optistat DN) and a temperature controller (Oxford Instruments ITC-502S). Emission spectrum for the ligand 1 was measured using a FP-6300 spectrofluorometer with a nitrogen bath cryostat (Oxford Instruments, Optistat DN) and a temperature controller (Oxford Instruments ITC-502S). Emission quantum yield for Eu(III) complex 2 was measured using a FP-6300 spectrofluorometer with an integration sphere (ILF-533).

NMR spectra were recorded in CDCl₃ using a JNM-EX 270 FT-NMR spectrometer (JEOL) for ¹H NMR experiments at 270 MHz and an AMX 500 (Bruker) for ¹H NMR experiments at 500 MHz. FD-MS and FI-MS analyses were performed on a JMS-T100GCV (JEOL) instrument. GC-MS analyses were performed using a Varian instrument. Authentic CJ was purchased from Sigma-Aldrich. LA-d5 (98%) was purchased from Cambridge Isotope Laboratories, Inc.

For the GC‐TOFMS analysis, we followed the method described in our previous publication (Kageyama et al., 2018). In brief, 100 µl of CSF samples was added to 25 µl of 1N HCL, 400 µl of methanol, and 1,000 µl of isooctane. Then, 2.5 ng of oleic acid‐d9 (Avanti Polar Lipids, AL, USA) was added to each CSF sample as an internal standard. The solution was thoroughly mixed and centrifuged at 800 × g for 2 min at room temperature. The supernatant was evaporated and then dissolved in 40 µl of pyridine, 40 µl of N‐(tert‐Butyldimethylsilyl)‐N‐methyltrifluoroacetamide, and 30 µl of acetonitrile for the GC‐TOFMS analysis by using JMS‐T100GCV (JEOL). The absolute concentration was quantified using a standard reference of nervonic acid (Sigma‐Aldrich).

The X-ray intensity data were measured on a Bruker D8 goniometer system equipped with a Bruker Turbo X-ray Source rotating-anode X-ray tube (MoKa, λ = 0.71073 Å) and a multilayered confocal mirror monochromator. ¹H- and ¹³C-NMR spectra were acquired on a Bruker Ascend 400 MHz NMR spectrometer (BRUKER BioSpin, Faellanden, Switzerland) at 400 and 125 MHz, respectively. LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) equipped with a source of electrospray ionization (ESI) was employed to get ESI-MS spectra of pure compounds. A Jasco HPLC system consisting of PU-4180 RHPLC pump, LC-Net II/ADC controller, and UV-4075 UV/Vis detector (Jasco, Tokyo, Japan) and the GC–MS system (JMS-T100 GCV, JEOL Ltd., Tokyo, Japan) were used to identify bioactive constituents. Biological activities were in vitro assayed by using a Multiskan^TM microplate spectrophotometer (Thermo Fisher Scientific, Osaka, Japan) and U-shape microplates (Greiner Bio-one, Monroe, NC, USA). Reagents, solvents, and chemicals at high grades were purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan), Junsei Chemical Co., Ltd. (Tokyo, Japan), Fisher Scientific company (Hampton, NH, USA) and Sigma-Aldrich (St. Louis, MO, USA).

A volume of 1 µL of sample was injected into a GC-MS system (JMS-T100 GCV, JEOL Ltd., Tokyo, Japan). The column was DB-5MS with 30 m in length, 0.25 mm internal diameter, and 0.25 µm in thickness (Agilent Technologies, J&W Scientific Products, Folsom, CA, USA). Helium was chosen as the carrier gas, and the split ratio was 5.0/1.0. The operating condition of GC oven temperature was maintained as follows: The initial temperature = 50 °C without hold time, the programmed rate = 10 °C/min up to a final temperature of 300 °C with 20 min for hold time. The injector and detector temperatures were set at 300 °C and 320 °C, respectively. The mass range scanned from 29–800 amu. The control of the GC-MS system and the data peak processing were carried out using the JEOL’s GC-MS Mass Center System version 2.65a software (JEOL Ltd., Tokyo, Japan).