Mat95xp

NMR spectra was recorded by a JEOL JEM-ECP NMR spectrometer (600 MHz for ¹H NMR and 150 MHz for ¹³C NMR). HRESIMS was tested on a Thermo MAT95XP high-resolution mass spectrometer. HRESIMS [M + Na]⁺m/z 295.1667 (calcd for C₁₈H₂₄O₂Na, 295.1669).

Optical rotations were measured in a JASCO DIP-1000 polarimeter (JASCO, Tokyo, Japan), with a Na (589 nm) lamp and filter. ¹H, ¹³C and 2D NMR spectra were recorded in a Bruker Avance 500 spectrometer, at 500 and 125 MHz, respectively, with CD₃OD and D₂O as solvents. HRESIMS experiments were performed in an Applied Biosystems QSTAR Elite system or a Thermo MAT95XP spectrometer. HPLC separations were performed in the Agilent 1100 liquid chromatography system equipped with a solvent degasser, quaternary pump, and diode array detector (Agilent Technologies, Waldbronn, Germany) with a semipreparative reversed phase column (Luna C18: 5 μ, 100 Å, 250 × 10 mm, Phenomenex, Lane Cove, Australia). Precoated silica gel plates (Merck, Kieselgel 60 F254, 0.25 mm, Merck Millipore, Merck KGaA, Darmstadt, Germany) were used for TLC analysis and the spots were visualized under a UV light (254 nm) or by heating the plate pretreated with H₂SO₄/H₂O/AcOH (1:4:20).

Low and high resolution mass spectra were obtained by staff at the University of Manchester. Electrospray (ES) spectra were recorded on a Waters Platform II with an SQ Detector 2. High resolution mass spectra (HRMS) were recorded on a Thermo Finnigan MAT95XP and are accurate to ±0.001 Da.

Low-resolution mass spectra EI-MS were chronicled on a JEOL MS route JMS 600H instrument, and HR-EI-MS was analyzed on Thermo Finnigan MAT 95XP linked with X-Calibur. The ¹H and ¹³C NMR spectra were recorded on a Bruker Avance NEO-500, 400 NMR spectrometer in CDCl₃ at 500, 400, and 125 MHz, respectively. The UV was checked on the Evolution^TM 300 Spectrophotometer, and FT-IR spectra were recorded on a Bruker Vector 22 spectrophotometer. Optical rotations were determined on a JASCO 2000 Polarimeter. The purity of the compounds was verified on TLC (Silica gel, Merck F254, 0.25 mm thickness). Melting points were determined in glass capillary tubes using the Buchi melting point apparatus. For the TLC plate’s visualization, vanillin and ceric sulfate staining reagents were used. All experiments were performed at room temperature using solvents acquired commercially and used without further purification.

Column chromatography (CC) was performed using silica gel (230–400 mesh, E. Merck, Darmstadt, Germany). Thin-layer chromatography (TLC) separations were carried out on pre-coated silica gel sheets (60 F_254. E. Merck), and the compounds were detected under UV light at 254 and 366 nm. Then, the plates were sprayed with ceric sulfate in 10% H₂SO₄ and were gently heated using a heat gun until the development in colour was observed. Recycling preparative HPLC (RP-HPLC (JAI LC-908W, Japan Analytical Industry Co. Ltd., Tokyo, Japan) with a YMC ODS H-80 or L-80 column (YMC, Tokyo, Japan) was used for the final purification. A UV-3200 spectrophotometer (Tokyo, Japan) was used to obtain the UV spectra, and the IR spectra were obtained from KBr discs on an A-302 spectrometer (JASCO, Tokyo, Japan). The EI-MS spectra were acquired on a EI (LR) JEOL MS ROUTE 600H, (JEOL Ltd, Tokyo, Japan). HREI-MS were recorded on a EI (HR) MAT 95XP ThermoFinnigan, Germany. The NMR spectra were acquired on AV-300, AV-400, and AV-500 instruments (Bruker, Switzerland). All chemical shifts values are displayed in ppm (δ). The ¹H-NMR, COSY, HSQC, and HMBC spectra were recorded at 400 MHz, while the ¹³C NMR spectra were obtained at 100 MHz. The coupling constants (J) are estimated in Hertz.

Mass spectra were recorded at the School of Chemistry, University of Manchester using the Micromass PLATFORM II (ES) and Thermo Finnigan MAT95XP (Accurate mass) instruments. Purification of products by semi-preparative HPLC was performed using a Shimadzu Prominence HPLC system controlled by Laura 3.0 software from LabLogic (Sheffield, UK). The system was configured with a CBM-20A controller, LC-20AB solvent delivery system, SPD-20A absorbance detector, FCV-20AH2 switching valve and Rheodyne^® 7725 injector. Radioactivity eluted by the HPLC system was monitored using a radio-HPLC Bioscan Flow Count B-FC 3100 detector. Radio-HPLC for quality control analysis was performed using a Shimadzu Prominence HPLC system configured with a CBM-20A controller, LC-20AB solvent delivery system, SPD-20A absorbance detector, SIL-20AHT autosampler and Bioscan Flow Count B-FC 3100 detector. Radioactivity measurements were made using an Isomed 2010 Dose Calibrator (MED Nuklear-Medizintechnik, Dresden, Germany).