Ecs 400 nmr spectrometer

NMR spectroscopy for the 5F-MDMB-PINACA, AB-FUBINACA and AMB-FUBINACA enantiomers was performed using a JEOL ECS-400 NMR spectrometer (JEOL, Tokyo, Japan) operating at 400 MHz. ¹H-NMR (10 mg/mL in CDCl₃), ¹³C-NMR (20 mg/mL in CDC_L3). NMR spectroscopy for the AB-CHMINACA enantiomers was performed using a Bruker AVANCE III HD 500 MHz spectrometer (Bruker, Billerica, MA, USA) running under TopSpin v.3.2.5 and equipped with a QCI-F cryo-probe at a sample compartment temperature of 25°C. Samples were prepared in CDCl₃ (~10 mg/mL).

The kinetic study along with a characterization of photo metabolite was carried out in Alliance 2695 Separations Module (Waters, Milford, MA, USA) attached with Micromass Quattro Micro triple-quadruple mass spectrometer (Micromass, Manchester, UK) using electrospray ionization in the positive ion (ES+) mode. IR spectra for the characterization of products were analyzed using KBr pellets (1.0 mm) using an infrared spectrophotometer (Make: Perkin Elemer, Model: Spectrum One, Model No: L120-000A). ¹H NMR spectra were obtained on a JEOL ECS-400 NMR spectrometer using TMS as the internal standard. X-ray crystallographic analysis was done on a Bruker SMART APEXII CCD area-detector diffractometer using graphite monochromatic Mo Kα radiation (λ = 0.71073 Å). X-ray data reduction was carried out using the Bruker SAINT program. The structures were hypothesized by direct methods using the SHELXS-97 program and refinement using SHELXL-97 program. The samples were centrifuged using a high-speed refrigerated centrifuge, Model Avanti J-30I (Beckman coulter, Brea, CA, USA). The rotor head was suitable for holding eight no. of 50 mL (JA-30.50 T1) fluorinated ethylene propylene (FEP) centrifuge tubes (Nalgene, Rochester, NY, USA). Samples were evaporated using a Turbo Vap LV instrument from Caliper Life Science (Hopkinton, MA, USA).

The liquid-state ¹³C-NMR spectra of the freeze-dried phenolic resins were recorded on a JEOL ECS-400 NMR spectrometer at a frequency of 75.51 MHz. DMSO-_D6 was used as the solvent. All of the spectra were recorded at room temperature with a relaxation delay of 8 s over 800 scans. The chemical shifts of each spectrum were accurate to 0.1 ppm and all the samples were directly used for ¹³C-NMR measurement.
The gel time of the phenolic resins was determined in accordance with Chinese National Standard (GB/T 14074.3-2006). Thermogravimetric measurement of the freeze-dried phenolic resins was performed with a Q50 thermogravimetric analyzer (TGA, TA Instruments, New Castle, DE, USA) in a nitrogen atmosphere (flow rate = 60 mL/min) and at a temperature range from room temperature to 600 °C, with a heating rate of 10 °C/min.

Thebaine, purchased from Sigma Aldrich (Poole, UK), was analysed using ¹³C and ¹H NMR. ¹H NMR spectra were obtained using a JEOL ECS 400 NMR spectrometer operating at a frequency of 400 MHz, using 8–32 scans, a relaxation delay of 5 s, and a flip angle of 45° (5 µs pulse). Spectra were Fourier transformed typically into 32 000 data points using standard exponential window with a line broadening factor of 0.2 Hz. ¹³C spectra were obtained at a frequency of 100.53 MHz, from 128–1048 scans, a relaxation delay of 2 s, and a flip angle of 30° (2.7 µs pulse). Spectra were Fourier transformed typically into 64 000 data points using standard exponential window with a line broadening factor of 0.5 Hz. Two-dimensional spectra (COSY, HSQC and HetCor) experiments were conducted using standard JEOL automated acquisition and processing parameters. Thebaine standards for analysis by NMR spectroscopy were prepared by dissolving approximately 10 mg of the powdered Thebaine standard into a 1 cm³ volume of the relevant deuterated solvent or solvent mixture incorporating 1% of deutero-acetic acid (CD₃COOD). Deuterated solvents were purchased from Goss Scientific (Crewe, UK).

¹H- and ¹³C-NMR spectroscopy of the carbon dioxide reduced liquid products were recorded a on a Bruker Ascend 700 MHz Avance III NMR spectrometer equipped with a cryoprobe and on a JEOL ECS-400 NMR spectrometer. As internal standard, 20 mL aqueous solution of 20 mM phenol and 10 mM of dimethyl sulfoxide were used. After CPE, to 350 µL electrolyte, 200 µL D₂O, and 50 µL of the internal standard were added and transferred into a NMR-tube. During the measurements, the water peak was suppressed to increase the signal intensity of the analytes. The CO₂ reduced products were further analyzed using GC–MS. Trace 1300 GC and ISQ QD single quadruple GC–MS instrument with a TG-5MS capillary column (30 m × 0.32 mm × 0.25 µm) supplied by Thermo Fisher Scientific and DB-624 capillary column (30 m × 0.32 mm × 0.25 µm) supplied by Agilent were used for the same. For gaseous analysis CarboPLOT 007 capillary column (25 m × 0.53 mm × 0.25 µm) supplied by Agilent was used for separation and TCD for detection.

All the chemicals and solvents were used from chemical suppliers without purification. Column chromatography was carried out for purification using precoated silica gel plates (MERCK silica gel 60; F254, 0.040–0.063 mm). NMR spectra were obtained using a JEOL ECS 400 NMR spectrometer (Tokyo, Japan) at a 1H frequency of 400 MHz and 13C frequency of 100 MHz. Proton and carbon chemical shifts were reported in parts per million (ppm) relative to an internal standard. Chemical shifts, multiplicities, and coupling constants (J) were reported and calculated using Delta 5.3.1 software provided by JEOL and ACD NMR processor academic edition software. Mass spectrometry of all synthesized compounds was carried out on a BEH C18 column (1.7 μm, 2.1 mm × 50 mm; Waters, Milford, MA, USA) maintained at 40 °C during separation under isocratic conditions (mobile phase A/mobile phase B = 20:80) using a Waters ACQUITY ultraperformance liquid chromatograph coupled to a triple quadrupole mass spectrometer (Micromass Quattro Micro, Waters). Mobile phase A consisted of water (LC–MS grade) with 0.1% formic acid (v/v) and mobile phase B consisted of CH3CN (LC–MS grade) with 0.1% formic acid (v/v); flow rate, 0.2 mL/min.