Avance 3 hd nanobay spectrometer

All reagents and solvents were purchased from commercial suppliers and were used directly without further purification. All reactions were carried out at room temperature unless otherwise specified. Reactions were monitored by thin-layer chromatography (TLC) on Merck F₂₅₄ silica gel 60 aluminum sheets, and spots were revealed with UV light (254 mm), potassium permanganate or Ninhydrin stains. ¹H NMR spectra were recorded on a 400 MHz Bruker Avance III HD Nanobay Spectrometer with deuterated chloroform (CDCl₃; δ = 7.24 ppm) or deuterated dimethyl sulfoxide (DMSO-d₆) containing TMS as internal standard. ¹³C NMR spectra were recorded on a Bruker Avance III HD Nanobay Spectrometer at 100 MHz. The percent purity of the inhibitors reported in this manuscript were determined by HPLC-UV using Agilent 1200 series HPLC system equipped with Phenomenex Luna2 C18 reverse phase column (C18, 4.6 mm × 150 mm, 5 μm) coupled with Agilent G1314 UV–vis detector (detection at 200, 210, 254 and 360) with solvent gradient acetonitrile/water 20 to 95% over 15 min and they are expressed as percent total OD at 254 and 360 nm. The purity of all final compounds was above 95%. HRMS spectra were recorded on Thermo Electron LTQ-Orbitrap XL Hybrid MS in ESI mode.

Chemicals were purchased and used without further purification. Nuclear magnetic resonance (NMR) spectra were recorded in the NMR facility of the University of California, Davis on a 800 MHz Bruker Avance III-NMR spectrometer or a 400 MHz Bruker Avance III HD Nanobay spectrometer. Chemical shifts are reported in parts per million (ppm) on the δ scale. High resolution electrospray ionization (ESI) mass spectra were obtained using a Thermo Scientific Q Exactive HF Orbitrap Mass Spectrometer at the mass spectrometry facility in the University of California, Davis. Column chromatography was performed using a CombiFlash^® Rf 200i system with either RediSep Rf silica columns or an ODS-SM (C18) column (51 g, 50 μm, 120 Å, Yamazen) or manually using columns packed with silica gel 60 Å (230–400 mesh, Sorbent Technologies). Thin layer chromatography (TLC) was performed on silica gel plates (Sorbent Technologies) using anisaldehyde sugar stain or 5% sulfuric acid in ethanol stain for detection. Gel filtration chromatography was performed with a column (100 cm × 2.5 cm) packed with Bio-Gel P-2 Fine resins (Bio-Rad). PmAldolase,^{31 (link)} NmCSS,^{32 (link)} PmST1,^{33 (link)} and Pd2,6ST^{34 (link)} were expressed and purified as described previously. Man2,4diN₃ (1) and Man2,4,6triN₃ (2) were synthesized as we previously reported.^{28 (link)}