Supporting Information for:
Ion Mobility Mass Spectrometry Analysis of
Isomeric Disaccharide Precursor, Product and Cluster Ions
Hongli Li1, Brad Bendiak2, William F. Siems1, David R. Gang3, and Herbert H. Hill, Jr.1, *
1
Department of Chemistry, Washington State University, Pullman, Washington, USA
2
Department of Cell and Developmental Biology, Program in Structural Biology and Biophysics, University of
Colorado, Health Sciences Center, Anschutz Medical Campus, Aurora, Colorado, USA
3
Institute of Biological Chemistry, Washington State University, Pullman, Washington, US
* To whom Correspondence should be addressed.
Herbert H. Hill, Jr., Department of Chemistry, Washington State University,
Pullman, WA 99164, USA email: hhhill@wsu.edu, Tel 509-335-5648
S-1
Table S-1: Additional Instrumental Parameters of Synapt G2 HDMS
Parameters
Sample Cone Voltage (V)
Extraction Cone Voltage (V)
Source Temperature (°C)
Helium Gas Flow Rate
(IMS mode)
Argon Gas Flow Rate
(Trap &Transfer cell)
Positive mode
40.0
4.0
150
180 mL/min
Negative mode
40.0
4.0
150
180 mL/min
2 mL/min
2 mL/min
Precursor Ion
[M+Na]+ at m/z 365.1
x102
0
0
2 2.2 2.4 2.6 2.8 3
2.2
x 105
0.1
0
0.8
0
2 2.2 2.4 2.6 2.8 3
0.5
2 2.2 2.4 2.6 2.8 3
x104
x106
0.8
0
3 3.2 3.4 3.6 3.8 4
Drift Time (ms)
β-D-Gal-GA
0
3 3.2 3.4 3.6 3.8 4
D-Gal-β-(1-6)D-Gal
β-D-Glc-GA
0
3 3.2 3.4 3.6 3.8 4
x 104
D-Gal-β-(1-4)D-Glc
x106
D-Glc-β-(1-2)D-Glc
b
4.4
3 3.2 3.4 3.6 3.8 4
x107
D-Glc-α-(1-4)D-Glc
x106
a
Product Ion
[M+Na]+ at m/z 245.1
α-D-Glc-GA
2.4
0.8
β-D-Gal-GA
0
2 2.2 2.4 2.6 2.8 3
Drift Time (ms)
Figure S-1. (a) TWIMS of isomeric disaccharide [M+Na]+ precursor ions at m/z 365.1. The disaccharides
displayed were D-Glc-α-(1-4)-D-Glc, D-Glc-β-(1-2)-D-Glc, D-Gal-β-(1-4)-D-Glc and D-Gal-β-(1-6)-D-Gal. (b)
TWIMS of Hex-glycolaldehyde product ions derived from disaccharides shown in (a) the positive mode
as [M+Na]+ ions at m/z 245.1. The corresponding product ions generated were sodiated α-D-Glc-GA, β-DGlc-GA, β-D-Gal-GA and β-D-Gal-GA.
S-2
x106
x200
0
x103
0.7
2
0
1.8
0
x104
x104
0.8
0
2.8 3 3.2 3.4 3.6 3.8
3
3.5
4
β-D-Glc-GA
0
2
2.5
3 3.5 4
β-D-Gal-GA
2
2.5
3 3.5 4
β-D-Gal-GA
0.3
0
2.8 3 3.2 3.4 3.6 3.8
D-Gal-β-(1-6)D-Gal
2.5
2.5
2.8 3 3.2 3.4 3.6 3.8
x105
D-Gal-β-(1-4)D-Glc
x103
D-Glc-β-(1-2)D-Glc
0
b
Product Ion
[M-H]- at m/z 221.1
α-D-Glc-GA
0.2
2.8 3 3.2 3.4 3.6 3.8
x103
D-Glc-α-(1-4)D-Glc
x104
a
Precursor Ion
[M-H]- at m/z 341.1
0.8
0.6
x4
0
2
2.5
3
3.5
4
Figure S-2. (a) TWIMS of isomeric disaccharide [M-H]- precursor ions at m/z 341.1. The disaccharides
displayed were D-Glc-α-(1-4)-D-Glc, D-Glc-β-(1-2)-D-Glc, D-Gal-β-(1-4)-D-Glc and D-Gal-β-(1-6)-D-Gal. (b)
TWIMS of Hex-glycolaldehyde product ions derived from disaccharides shown in (a) in the negative
mode as [M-H]-ions at m/z 221.1. The corresponding product ions generated were deprotonated α-DGlc-GA, β-D-Glc-GA, β-D-Gal-GA and β-D-Gal-GA.
S-3
5.5
00
00
2.8
3 3.2
5 5.5
6 3.4
6.53.67 3.87.54
DriftTime
Time (ms)
Drift
(ms)
6
6.5
7
6
6.5
7
7.5
7.5
1.8
0
5.5
6
6.5
7
(ms)
Drift Drift
TimeTime
(ms)
0.5
0
7 7.5 8 8.5 9 9.5 10
7.5
0
5.5
x 100000
x106
Thousands
D-Glc
1.8
1.2
7
2.2
2.8
3 3.2
5 5.5
6 3.4
6.53.67 3.87.54
D-Gal-β-(1-4)Gal-β-1-4-Glc
6.5
0
5.5
x 100000
x104
x 10000
D-Glc
0.3
0.6
6
3.2
2.8
3 3.2
5 5.5
6 3.4
6.53.67 3.87.54
Glc-β-1-4-Glc
D-Glc-β-(1-4)-
x 10000
0
Thousands
x 100000
x 10000
1.2
Thousands
00
[3M-H]
m/z1025.4
1025.3
c c[3M-H]
ion- at m/z
x 10000
D-Gal
0.8
0.6
- - at m/z 683.2
[2M-H]
bb[2M-H]
ion at m/z 683.2
x 10000
D-Gal-β-(1-6)Gal-β-1-6-Gal
00
5 5.5
6 3.4
6.53.67 3.87.54
2.8
3 3.2
x104
Gal-α-16-Glc
D-Glc
0.1
1.2
x 10000
D-Gal-α-(1-6)-
x106
+ at m/z 707.2
a [2M+Na]
a [M-H]- ion
at m/z 341.1
7.5
1.8
0
7 7.5 8 8.5 9 9.5 10
1.6
0
7 7.5 8 8.5 9 9.5 10
0.1
0
7 7.5 8 8.5 9 9.5 10
Time
(ms)
DriftDrift
Time
(ms)
Figure S-3. Traveling wave ion mobility separation (TWIMS) of isomeric disaccharide cluster ions of (a)
[2M+Na]+ ions at m/z 707.2 (b) [2M-H]- ions at m/z 683.2 and (c) [3M-H]- ions at m/z 1025.3. The
disaccharides used are labeled at the left in the same order from top to bottom for all three panels.
S-4
neg sen, 50uM with 100uM NH4CL
neg sen, 50uM with 100uM NH4CL
%
379.1
380.1
255.2
0
200
0
200
%
%
%
%
377.1
%
100
400
%
377.1
100
%
400
neg sen, 50uM with 100uM NH4CL
%
255.2
0
200
1061.3
%
%
379.1
380.1
3E4
1063.3
Gal-α-1-6-Gal 1049.3
1061.3
0
719.1 100
1063.3 1070 m/z
1050 1055 1060 1065
683.2
1049.3
1061.3
100
0
m/z m/z
1063.3
600
800 1050 1055 1000
1200
1060
1065
1070
1061.3
1049.3
100
1063.3
0
m/z
1050 1055 10601061.3 1065 4E4
1070
0
100
1063.3 1070 m/z
1050 1055 1060 1065
Glc-β-1-4-Glc
1061.3
100
0
m/z
1050 1055 1061.2
1060 1063.3
1065 1070
719.2
100
1063.2
m/z
0
m/z
600
800
1000
1050 1055
1053.2 1060
1061.2 1065 10701200
100
0
1063.2 1070 m/z
1050 1055 1060 1065
2E4
1053.2 1061.2
0
100
1063.2
Gal-β-1-4-Man 1050 1055 1060 1065 1070 m/z
1053.2
0
m/z
719.11050 1055 1060 1065 1070
m/z
600
800
1000
1200
neg sen, 50uM with 100uM NH4CL
100
%
377.1
100
379.0
311.1
400
Figure S-4. (a) The mass spectra of disaccharides as chloride adducts of [M+Cl]- ion at m/z 377.1,
[2M+Cl]- ion at m/z 719.1 and [3M+Cl]- ion at m/z 1061.2 in the negative mode. The representative
examples are D-Gal-α-(1-6)-D-Glc, D-Glc-β-(1-4)-D-Glc and D-Gal-β-(1-4)-D-Man from top to bottom. Mass
accuracy was within ± 0.1 m/z units in this investigation.
S-5
x 105
D-Gal-α-(1-6)D-Glc
b [2M+Cl]- ion at m/z 719.1
1.6
0
0
x 104
×106
0
6.5
8
9
10
7
8
9
10
7
8
9
10
7
8
9
10
0.4
0
2.5
0
6.5
Drift Time (ms)
0
7.5
x 104
x 105
7
4.5
5.5
Drift Time (ms)
0
7.5
0.6
5.5
D-Gal-β-1-4D-Glc
6.5
0.6
7.5
0.5
5.5
D-Glc-β-(1-4)D-Glc
6.5
x 104
x 105
5.5
D-Gal-β-(1-6)D-Gal
c [3M+Cl]- ion at m/z 1061.2
x 104
a [M+Cl]- ion at m/z 377.1
7.5
1.4
0
Drift Time (ms)
Figure S-5. (a) TWIMS of the selected disaccharide [M+Cl]- ions at m/z 377.1. (b) TWIMS of the same
selected disaccharide [2M+Cl]- ions at m/z 719.1. (c) TWIMS of the same selected disaccharide [3M+Cl]ions at m/z 1061.2. The isomeric disaccharides shown at the left are in the same order from top to the
bottom for all three panels.
S-6