An aliquot of each purified LPS was treated with acetate buffer (pH 4.4, 2 h, 100 °C) in order to separate the lipid A portion from the saccharide part of the LPS. A mixture of chloroform and methanol was added to the hydrolysis product to obtain a CHCl3/CH3OH/hydrolysate 2:2:1.8 (v/v/v) ratio. The mixture was then shaken and centrifuged. The chloroform phase, containing the lipid A, was collected and washed with the water phase of a freshly prepared Bligh/Dyer mixture (CHCl3/CH3OH/H2O, 2:2:1.8) [21 (link)]. The organic phases were pooled, dried, and analyzed by MALDI-TOF MS (SCIEX, Concord, ON, Canada). In order to establish the nature of the sugar backbone of each lipid A, an aliquot of each lipid A fraction also underwent a methanolysis (1.25 M HCl/CH3OH, 80 °C, 16 h) followed by acetylation (80 °C, 20 min) and GC-MS analysis [22 (link)].
Maldi tof ms
Manufactured by AB Sciex
Sourced in Canada
The MALDI-TOF MS is a mass spectrometry instrument that uses Matrix-Assisted Laser Desorption/Ionization (MALDI) to ionize and analyze samples. It is designed to determine the masses of molecules in a sample, providing information about their molecular weights and compositions.
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Lab products found in correlation
2 protocols using maldi tof ms
1
Lipid A Structural Analysis of LPS
2
Spectroscopic Analysis of Organic Compounds
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All chemicals and solvent were of pure analytical grade. Thin layer chromatography (TLC) was executed on 60 F-254 silica gel, coated on aluminium plates and visualized by UV lamp, 5% H2SO4 methanol solution with subsequent charring by heating. In addition, alkaline KMnO4 solution was used to locate the alkynes with subsequent heating. Flash column chromatography was performed using silica gel (230–400) mesh size. 1H and 13C spectra were recorded in 500 and 125 MHz respectively. Mass spectra were recorded on Sciex-X500R QTOF, MALDI-TOF MS (with 2,5-dihydroxy benzoic acid matrix). IR was recorded in Nujol mulls in KBr pellets. The polydispersity index (Mn/Mw) of the samples was investigated in DMF at 40 °C with a flow rate of 0.5 mL min−1. The calibration of columns was done against polystyrene standard samples. The absorbance and emission spectra were also recorded by using UV-visible spectroscopy.
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