Electronic Supplementary Material (ESI) for ChemComm.

This journal is © The Royal Society of Chemistry 2015

Supramolecular chemical shift reagents inducing

conformational transition: NMR analysis of carbohydrate
homooligomer mixtures
Sophie  R.  Beeren*a  and  Sebastian  M eier* b  

Supporting Information

Contents

S1 Materials S1
S2 NMR spectroscopy S3

S1 Materials

Glucose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, 8-hydroxypyrene-

1,3,6-trisulfonic acid trisodium salt (HPTS) and sodium dodecyl sulfate (SDS) were obtained
from Sigma–Aldrich. Maltoheptaose and maltooctaose were obtained from Carbosynth
(Compton, UK). D2O was purchased from Cambridge Isotope Laboratories, (Andover, MA,
USA). HPTS-C16, HPTS-C12, HPTS-C8 and HPTS-C4 were synthesized as described
previously.1,2 The maltooligosaccharide mixture was prepared by fractional precipitation of a
starch digest with ethanol as described previously.3 The distribution of maltooligosaccharides
in the mixture was determined using UPLC/MS analysis following fluorescence labelling at
the reducing end with 2-aminobenzamide as described previously.2

S1
 DP 5

DP 10

DP 15

DP 20

Figure S1. Fluorescence chromatogram showing UPLC analysis of the α(1-4) glucan
mixture used in this work.

S2
S2 NMR spectroscopy

1
H-13C HSQC NMR spectra were recorded on an 800 MHz Bruker (Fällanden, Switzerland)
Avance spectrometer equipped with a 5 mm TCI z-gradient CryoProbe and an 18.7 T magnet
(Oxford Magnet Technology, Oxford, UK) and processed with Topspin 3.0 (Bruker) using
extensive zero filling in both dimension, no linear prediction and mild resolution
enhancement in the 13C dimension. Sensitivity enhanced 1H-13C HSQC spectra were acquired
with narrow spectral width in the indirect dimension using the standard Bruker pulse
sequence (hsqcetgpsi). Specifically:

For analysis of the α-glucan mixture with 10 mM HPTS-C16, SDS or HPTS

HSQC spectra were acquired as matrices of 1024×256 complex data points sampling 143
milliseconds in the direct (1H) and 319 milliseconds in the indirect (13C) dimension. The
spectra were acquired with a spectral width of 9 ppm (7183 Hz) in the direct dimension and of
4 ppm (803 Hz) in the indirect dimension with a recycle delay of 1 second. For analysis of the
α-glucan mixture alone and in the presence of HPTS, 2 scans were accumulated. The resolved
spectra of the α-glucan mixture in the presence of 10 mM HPTS-C16 and 10 mM SDS were
recorded over night with 96 scans.
One-dimensional 1H NMR spectra of the mixture alone and in the presence of 10 mM
SDS, HPTS and HPTS-C16 are shown in Figure S2. Spectra were acquired at 800 MHz by
sampling 16384 complex data points for an acquisition time of 1.27 seconds and employing a
spectral width of 12820 Hz. 1H NMR spectra were processed with extensive zero filling and
with an exponential line broadening of 0.3 Hz.

For analysis of the α-glucan mixture together with HPTS-C12, HPTS-C8 and HPTS-C4:
HSQC spectra were acquired as matrices of 1024×192 complex data points sampling 143
milliseconds in the direct (1H) and 319 milliseconds in the indirect (13C) dimension by
accumulating 96 scans for HPTS-C12 and HPTS-C8 with a recycle delay of 1 second and by
accumulating 76 scans in the case of HPTS-C4 with a recycle delay of 1 second.

For assignment of signals using commercial reference compounds:

Mixtures of G1, G2, G3, G6 and G8 (each 2 mg/ml) and of G2, G4, G5, G6 and G7 (each 2
mg/ml) were prepared and subjected to HSQC spectroscopy for the validation of mixture

S3
signal assignments by the use of reference compounds. Both mixtures were analysed in the
presence of 10 mM (i) HPTS-C4, (ii) HPTS-C8, (iii) HPTS-C12 or (iv) HPTS-C16 (Figure S5).
The HSQC spectra were acquired as matrices of 1024×256 complex data points sampling 143
milliseconds in the direct (1H) and 319 milliseconds in the indirect (13C) dimension. Spectra
were recorded with a spectral width of 9 ppm (7183 Hz) in the direct dimension and of 4 ppm
(803 Hz) in the indirect dimension with a recycle delay of 1 second and 8 scans.

α(1-4) glucan mixture

(a)

(b) +10 mM SDS

(c) +10 mM HPTS

(d) +10 mM HPTS-C16

8 6 4 2

Chemical Shift (1H, ppm)

Figure S2. 1H NMR spectra (300 K, 800 MHz, D2O) of the α(1-4) glucan mixture alone (a)
and in the presence of 10 mM SDS (b), HPTS (c), and HPTS-C16 (d). Anomeric regions of the
corresponding HSQC spectra are shown in panels a-d of the main text Figure 2.

S4
 H1

H1β-red
H1α-red
H2β-red

H5β-red

H1β-red
H1α-red

H1

H2β-red

H5β-red

Figure S3. 1H13C HSQC spectra (300K, 800 MHz, D2O) of the α(1-4) glucan mixture alone
(a) and in the presence of 10 mM HPTS-C16 (b). The spectra were recorded with a narrow (2
and 4 ppm) sweep width in the 13C dimension and therefore many signals are multiply
aliased.

S5
(a)

(b)

Figure S4. 1H13C HSQC spectra (300 K, 800 MHz, D2O) showing (a) H1β reducing end
signals and (b) the H5β reducing end signals of the 10 mg/ml α(1−4) glucan mixture in the
presence of 10 mM of (i) HPTS-C4, (ii) HPTS-C8, (iii) HPTS-C12 and (iv) HPTS-C16.

S6
(a)

(b)

(c)

Figure S5. 1H13C HSQC spectra (300 K, 800 MHz, D2O) showing (a) the H1β reducing end
signals, (b) the H2β reducing end signals and (c) the H5β reducing end signals of: in red, a
mixture of G1, G2, G3, G6 and G8 (each 2 mg/ml) and in blue, a mixture of G2, G4, G5, G6 and
G7 (each 2 mg/ml) in the presence of 10 mM (i) HPTS-C4 or (ii) HPTS-C8, (iii) HPTS-C12
and (iv) HPTS-C16.

1
Beeren, S. R.; Hindsgaul, O. Angew. Chem. Int. Ed. 2013, 52, 11265-11268.
2
Beeren, S. R.; Meier, S.; Hindsgaul, O. Chem. Eur. J. 2013, 19, 16314-16319.
3
Johannesen, S. A.; Beeren, S. R.; Blank, D.; Yang, B. Y.; Geyer, R.; Hindsgaul, O. Carbohydr. Res. 2012, 352,
94-100.  

S7