Agilent 1260 infinity quaternary lc apparatus

UV spectra were recorded on a Jasco V-530 spectrophotometer (Jasco Europe s.r.l., Cremella, Italy); ECD spectra were recorded on a Jasco 715 spectropolarimeter using 1 cm cuvettes; ¹H NMR and 2D NMR experiments were carried out at 700 MHz on a Bruker Avance Neo spectrometer (Bruker BioSpin Corporation, Billerica, MA, USA) equipped with a cryoprobe for standard 5 mm tubes; ¹H and ¹³C chemical shifts were referenced to the residual solvent signal (CD₃OD, δ_H 3.31, δ_C 49.0); ³¹P chemical shifts were referenced to aqueous 85% w/w H₃PO₄ (δ_P = 0 ppm) as an external standard. The multiplicity-edited HSQC spectra was optimized for ¹J_CH = 145 Hz and the HMBC experiments for ^2,3J_CH = 8.0 Hz and ^2,3J_PH = 8.0 Hz. Through-space ¹H connectivities were evidenced using a ROESY experiment with a mixing time of 200 ms. High-resolution ESI-MS and HR-ESI-HPLC experiments were recorded on a Thermo LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific Inc., Waltham, MA, USA) combined to a Thermo U3000 HPLC system. High-performance liquid chromatography (HPLC) separations were achieved on an Agilent 1260 Infinity Quaternary LC apparatus (Agilent Technology, Cernusco sul Naviglio, Italy), equipped with a diode-array detector (DAD).

NMR spectra were determined on Varian Unity Inova spectrometers at 700 MHz and 500 MHz. Chemical shifts were referenced to the residual solvent signal (CD₃OD: δ_H 3.31, δ_C 49.0). For an accurate measurement of the coupling constants, the one-dimensional ¹H NMR spectra were transformed at 128 K points (digital resolution < 0.1 Hz). The HSQC spectra were optimized for ¹J_CH = 142 Hz and the ¹³C HMBC experiments for ^2,3J_CH = 8.0 Hz. Medium pressure liquid chromatography were performed on a Buchi Gradient System C-605 apparatus using glass columns of LiChroprep RP-18 (25–40µm) and a C-660 Buchi fraction collector. TLC performed on SiO₂ plates with BuOH/H₂O/CH₃CO₂H 60:25:15 (BAW) as a mobile phase and cerium sulphate in 2 N H₂SO₄ as a reagent for visualizing the spots. High performance liquid chromatography (HPLC) separations were achieved on a Waters 515 apparatus equipped with a refractive index detector (Waters 2414) using semi-preparative Novapak C18 (300 mm × 7.8 mm i.d.) and analytical Novapak C18 (300 mm × 3.9 mm i.d.) columns. Final purification were performed on an Agilent 1260 Infinity Quaternary LC apparatus (Agilent Technology, Cernusco sul Naviglio, Italy) equipped with a Diode-Array Detector (DAD).

A Jasco P-2000 polarimeter (Jasco Europe s.r.l., Cremella, Italy) at the sodium D line was used to measure optical rotations. ¹H NMR and 2D NMR experiments were carried out at 700 MHz on a Bruker Avance Neo spectrometer (Bruker BioSpin Corporation, Billerica, MA, USA) using dimethylsulfoxide-d₆ (DMSO-d₆) as solvent; all chemical shifts were referenced to the residual solvent signal (δ_H 2.50, δ_C 39.5). The HSQC spectra were optimized for ¹J_CH = 142 Hz and the HMBC experiments for ^2,3J_CH = 8.3 Hz. Through-space ¹H connectivities were evidenced using a NOESY experiment with a mixing time of 300 ms. High-resolution MS and LC-MS experiments were recorded on a Thermo LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific Inc., Waltham, MA, USA) combined to a Thermo U3000 HPLC system. High-performance liquid chromatography (HPLC) separations were achieved on an Agilent 1260 Infinity Quaternary LC apparatus (Agilent Technology, Cernusco sul Naviglio, Italy), equipped with a diode-array detector (DAD).