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2D heteronuclear multiple bond coherence (HMBC) played an important role in studying the stereoregularity of the carbonyl carbon.

Assignment of proton signals was achieved through two-dimensional (2D) experiments: homonuclear 1H-1H correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), nuclear Overhauser enhancement spectroscopy (NOESY), and heteronuclear 1H-13C heteronuclear single-quantum correlation (HSQC), and heteronuclear multiple bond coherence (HMBC).

DOI: http://dx.doi.org/10.7554/eLife.03941.018 The H and C resonances of the isolated tetrasaccharide product were assigned by using heteronuclear multiple quantum coherence (HMQC), heteronuclear 2-bond correlation (H2BC), and heteronuclear multiple bond coherence (HMBC) spectra.

Proton-detected heteronuclear correlations were measured using a gradient heteronuclear single-quantum coherence (HSQC), optimised for JHC = 155 Hz, and a gradient heteronuclear multiple bond coherence (HMBC), optimised for n JHC = 8 Hz.

The structure of this isolated compound was determined by a combination of spectroscopic analyses, including IR, H and C NMR, hydrogen-hydrogen correlation spectroscopy (H-H COSY), heteronuclear multiple quantum coherence (HMQC), heteronuclear multiple bond coherence (HMBC), mass spectroscopic data, and chemical methods.

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H-H-correlated spectroscopy (COSY), and heteronuclear multiple bonds coherence (HMBC) spectra were recorded on a 600 MHz Bruker DMX-600 spectrometer (Bruker).

Identification of these and other compounds was based also on correlation among specific signals given by H-H correlated spectroscopy (COSY) spectra (Additional File 4) and heteronuclear multiple bonds coherence (HMBC) spectra.

H NMR, 2D-J resolved, H-H correlated spectroscopy (COSY), and heteronuclear multiple bonds coherence (HMBC) spectra were recorded at 25°C on a 600 MHz Bruker AV 600 spectrometer equipped with cryo-probe operating at a proton NMR frequency of 600.13 MHz.

A metabolite detected as being more abundant in a specific sample can be identified either through the resonance positions of its nuclei in the NMR spectrum, or through the application of various pulse-sequences such as total correlation spectroscopy, heteronuclear single quantum coherence and heteronuclear multiple bond correlation.

Their structures were confirmed by 700 MHz Bruker, BioSpin nuclear magnetic resonance (NMR) analysis including one-dimensional 1H NMR, 13C NMR and two-dimensional NMR-correlation spectroscopy (COSY), hetero-nuclear single quantum coherence (HSQC), and heteronuclear multiple bond connectivity (HMBC).

The 1H NMR spectra of ADP-1a, ADP-1b, ADP-2, ADP-3a, and ADP-3b were recorded, 13C NMR spectra, the 2D NMR spectra including heteronuclear multiple-quantum coherence (HMQC) and heteronuclear multiple bond correlation (HMBC) of ADP-1a and ADP-2 were recorded.

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