Drx 400 nmr spectrometer

The molecular structure of the PSO extracted by UAE was analyzed using ¹HNMR. The oil sample was dissolved in the deuterated chloroform. ¹HNMR spectra of the extracted PSO were recorded using a Bruker DRX‐400 NMR spectrometer (Bruker Instruments) with ¹H nuclei observed at 400 MHz (Almoselhy, Allam, El‐Kalyoubi, & El‐Sharkawy, 2014).

A JASCO P-2000 digital polarimeter was utilized to measure the specific rotations of the compounds. A Bruker DRX-400 NMR spectrometer was utilized to obtain the NMR spectra of the compounds. Specifications of the NMR spectrometer include a Cryoprobe, and a 5 mm BBI (1H, G-COSY, multiplicity-edited G-HSQC, and G-HMBC spectra) or BBO (13C spectra) probe heads equipped with z-gradients. Residual solvent peaks for DMSO-d₆ were set at δ_H 2.50 and δ_C 39.5 ppm as reference signals in the ¹H and ¹³C NMR spectra, respectively. A preparative HPLC experiment was performed using Agilent 1260 Infinity Preparative-scale LC/MS Purification System coupled to an Agilent 6130B single quadrupole mass spectrometer with an XTerra Prep MS C₁₈ column (19 × 300 mm, 10 µm). The detection wavelength used in the preparative HPLC was 254 nm. An Agilent UHPLC 1290 Infinity, coupled with an Agilent 6540 accurate–mass quadrupole time-of-flight (QTOF) mass spectrometer, equipped with an ESI source was utilized to conduct the HPLC-MS experiment. The analyses were conducted with an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 µm), at a flow rate of 0.5 mL/min under standard gradient conditions of 2% MeCN (0.1% formic acid) to 100% MeCN (0.1% formic acid) over 8.6 min. The operating parameters for QTOF were the same as previously reported [8 (link)].

JASCO P-2000 digital polarimeter was used to record the optical rotations while GE Healthcare Ultrospec 9000 spectrophotometer was used to obtain the UV spectra.
Bruker DRX-400 NMR spectrometer with Cryoprobe was used to collect the NMR spectra. 5-mm BBI (¹H, G-COSY, multiplicity-edited G-HSQC, and G-HMBC spectra) or BBO (¹³C spectra) probe heads equipped with z-gradients were used.
Agilent 1260 Infinity Preparative-Scale LC/MS Purification System and Agilent 6130B single quadrupole mass spectrometer for LC and LC/MS Systems were used to perform the preparative HPLC analysis.
Agilent UHPLC 1290 Infinity coupled to Agilent 6540 accurate-mass quadrupole time-of-flight (QTOF) mass spectrometer which was equipped with a splitter and an ESI source were used to acquire the HRESIMS and MS/MS spectra. For over 15 min, under standard gradient condition of 100% water with 0.1% formic acid to 100% acetonitrile with 0.1% formic acid, the analysis was performed with a C18 4.6 × 75 mm, 2.7 μm column at flowrate of 2 mL/min. The operating parameters for QTOF were the same as in [6 (link)].
Nα-(2,4-Dinitro-5-fluorophenyl)-L-alaninamide (L-FDAA) and amino acid standards were purchased from Sigma Aldrich except D-allo-threonine which was purchased from Chem Cruz. Aspartic acid and glutamic acid were converted from asparagine and glutamine (Sigma Aldrich), respectively.

NMR spectra were performed on a Bruker DRX-400 NMR spectrometer at 400 MHz using CD₃OD as solvent in Zhengzhou University. The purity of morroniside and the derivatives were checked on a Waters e2695 liquid chromatograph equipped with 2424 ELS detector and 2998 PDA detector. GF254 silica plate was purchased from Qingdao Haiyang Chemical Co. Ltd. (Qingdao, China). The Elisa kits of OC (H152) and ALP (A059-2) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All the chemical reagents were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). The morroniside was extracted and purified in the medicinal chemistry department, Heilongjiang University of Chinese Medicine.

¹H and ¹³C-NMR spectra were recorded on a Bruker DRX-400 NMR spectrometer (¹H: 400 MHz; ¹³C: 100 MHz). Chromatography was performed on reversed-phase silica Kieselgel 60 silanisiert (0,063–0,200 mm). Eluates were monitored by thin-layer chromatography (TLC) on silica 60 F₂₅₄ (Merck) using butanol/acetic acid/water (BAW 8 : 1 : 1), visualized under UV light and revealed with NP-PEG.

The melting points were determined by an uncorrected WRX-4 binocular microscope (Shanghai Yice Tech. Instrument Co., Shanghai, China). ¹H-NMR and ¹³C-NMR spectral analyses were performed on a Bruker DRX-400 NMR spectrometer (Bruker, Rheinstetten, Germany). HRMS data were measured on an Agilent Technologies 6210 LC/MS TOF mass spectrometer (Agilent, Palo Alto, CA, USA). All reagent products from the Chinese Chemical Reagent Company were analytical or chemically pure.