SUPPORTING INFORMATION
Transmission mode direct analysis in real time mass spectrometry for fast
untargeted metabolic fingerprinting
Christina M. Jones and Facundo M. Fernández*
School of Chemistry and Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive
NW, Atlanta, GA 30332, USA
*Correspondence to: F. M. Fernández, School of Chemistry and Biochemistry, Georgia Institute
of Technology, 901 Atlantic Drive NW, Atlanta, GA 30332, USA.
E-mail: facundo.fernandez@chemistry.gatech.edu
S-1
Figure S1. Photograph of the automated TM-DART system. The top panel displays the module
used to hold the stainless steel mesh strip or discs, as well as a stainless steel mesh strip. The
bottom panel shows the automated TM system during an analysis placed between the DART ion
source and the GIST inlet.
S-2
8e+4
m/z 169.0675
m/z 289.1305
m/z 379.1799
m/z 500.2723
6e+4
Absolute Intensity
4e+4
2e+4
0
1.4e+5
m/z 586.3268
m/z 626.3538
m/z 730.3848
1.2e+5
1.0e+5
8.0e+4
6.0e+4
4.0e+4
2.0e+4
0.0
400
450
500
550
600
Time (sec)
Figure S2. Effect of ramping the set helium gas temperature on intensities of monitored mass
spectrometric signals in the low (top panel) and high (bottom panel) mass ranges. The set helium
gas temperature was ramped from 150 to 450°C over 3 min.
S-3
1.8e+5
1.4e+5
8e+5
Absolute Intensity
1.6e+5
Absolute Intensity
1e+6
m = 1.04e+3 ± 70
b = 3 ± 5e+3
r2 = 0.983
1.2e+5
1.0e+5
8.0e+4
6.0e+4
4.0e+4
2.0e+4
m = 2.8e+4 ± 6e+3
b = -8e+4 ± 9e+4
r2 = 0.924
6e+5
4e+5
2e+5
0
0.0
0
20
40
60
80
100
120
140
160
Quinine Concentration (µM)
0
5
10
15
20
Quinine Concentration (µM)
25
30
Figure S3. Signal linearity for TM-DART (left) and PM-DART (right). The absolute intensity of the [M+H]+ quinine ion was
monitored (n = 3) for 1 (PM), 10, 15, 25, 75 (TM), 100 (TM), and 150 (TM) µM solutions. The experimental data was linearly fitted
to a 𝑦 = 𝑚𝑥 + 𝑏 model. The regression parameters for each DART operational mode are displayed within each panel.
S-4
Intens.
TM-DART
103
102
0.5
Intens.
104
1.0
1.5
2.0
2.5
3.0
Time [min]
SS Mesh Discs_Quinine (Pyridine)_25uM_2.d: EIC 325.00000 +All MS
PM-DART
103
102
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
Time [min]
10TSM3D3_G_Pos3_04mmsec_1uL_2.d: EIC 195.0 +All MS
Figure S4. Extracted ion chronograms of the [M+H]+ (m/z 325.1920) ion observed during TM-DART (top panel) and PM-DART
(bottom panel) analysis of a 15 µM quinine solution.
S-5
Table S1. Tentative annotation of peaks selected from the chemically derivatized human blood
serum mass spectrum shown in Fig. 1 via accurate mass measurements with a mass tolerance of
5 mmu and a relative intensity cutoff of 1% of the base peak
Measur
ed
Ions
(m/z)
Ion Type
Experime
ntal
Exact
Mass (Da)
Theoreti
cal
Exact
Mass
(Da)
Accura
cy
(mmu)
S-6
Estimate
d
Formula
Name
Source
133.07
97
146.10
04
162.09
72
188.11
24
190.12
89
204.14
34
244.15
53
250.13
44
271.12
50
[M+TMS+
H]+
[M+TMS+
H]+
[M+TMS+
H]+
[M+TMS+
H]+
[M+TMS+
H]+
[M+TMS+
H]+
[M+H]+
315.10
50
364.17
57
389.14
29
[M+2TMS+
H]+
[M+2TMS+
H]+
[M+2TMS+
H]+
[M+3TMS+
H]+
[M+3TMS+
H]+
[M+3TMS+
H]+
[M+3TMS+
H]+
393.21
39
[M+2TMS+
H]+
453.23
53
598.29
42
600.30
74
[M+4TMS+
H]+
[M+5TMS+
H]+
[M+5TMS+
H]+
315.10
50
60.0323
60.0323
0.0
CH4N2O
73.0530
73.0527
0.3
C3H7NO
89.0499
89.0476
2.3
115.0651
117.0816
131.0961
243.1475
105.0475
126.0381
115.063
3
117.078
9
131.094
6
243.147
0
105.042
5
126.042
9
1.8
2.7
1.5
0.5
5.0
97.9786
97.9768
1.8
248.1270
164.0683
237.0887
239.1019
3.9
2.8
248.127
3
3.0
164.068
4
237.086
1
239.101
8
0.1
2.6
0.1
a
2
C5H11N
O2
C6H13N
O2
C12H21N
O4
C3H7NO
3
C7H6O5
3.4
172.0164
C5H9NO
C5H6N2
O2
170.021
5
147.053
1
172.013
6
2
4.8
170.0181
147.0492
C3H7NO
HMDB002
94
Aminoaceton HMDB021
e
34a
HMDB001
L-Alanine
61b
HMDB001
L-Proline
62c
HMDB008
L-Valine
83d
HMDB006
L-Leucine
87e
Tiglylcarniti HMDB023
ne
66
HMDB001
L-Serine
87f
HMDB002
Thymine
62g
Urea
Gallic acid
Phosphoric
acid
C5H9NO L-Glutamic
acid
4
Glycerol 3C3H9O6P
phosphate
Histidylproli
C12H16N4
ne
O2
diketopiperaz
ine
H3O4P
C6H12O5
C9H11N5
O3
C9H13N5
O3
HMDB058
07
HMDB021
42
HMDB001
48h
HMDB001
26i
HMDB020
53
HMDB001
74j
HMDB004
Biopterin
68k
Dihydrobiopt HMDB000
erin
38l
L-Fucose
One isomer found with source ID HMDB01106; bThree isomers found with source IDs HMDB00056,
HMDB00271, HMDB01310; cTwo isomers found with source IDs HMDB03411, HMDB12880; dThree isomers
found with source IDs HMDB13716, HMDB02141, HMDB03355; eFour isomers found with source IDs
HMDB01645, HMDB00557, HMDB03640, HMDB13773; fOne isomer found with source ID HMDB03406; gOne
isomer found with source ID HMDB02024; hTwo isomers found with source IDsHMDB06556, HMDB02931; iOne
isomer found with source ID HMDB02520; jSix isomers found with source IDs HMDB03081, HMDB05876,
HMDB00849, HMDB10207, HMDB12327, HMDB02712; kFive isomers found with source IDs HMDB01195,
HMDB02263, HMDB00633, HMDB00817, HMDB00238; lThree isomers with source IDs HMDB13642,
HMDB02215, HMDB02065.
S-7
Table S2. Expansion of tentative annotation of peaks given in Table S-1 selected from the
chemically derivatized human blood serum mass spectrum shown in Figure 1 via accurate mass
measurements with a mass tolerance of 10 mmu and a relative intensity cutoff of 1% of the base
peak
Measur
ed
Ions
(m/z)
146.10
04
Ion Type
Experime
ntal
Exact
Mass
(Da)
Theoreti
cal
Exact
Mass
(Da)
Accura
cy
(mmu)
Estimate
d
Formula
Name
Source
[M+H]+
145.0926
145.085
1
7.5
C5H11N3
O2
4Guanidinobutanoi
c acid
HMDB034
64
189.1211
189.111
3
9.8
C7H15N3
O3
Homocitrulline
HMDB006
79
59.0565
59.0483
8.2
CH5N3
Guanidine
HMDB018
42
Mevalonic acid
HMDB002
27a
204.14
34
[M+H]+
[M+2TMS
+H]+
[M+2TMS
+H]+
221.12
88
[M+TMS+
H]+
148.0814
148.073
5
7.9
C6H12O4
250.13
44
[M+TMS+
H]+
177.0870
177.078
9
8.1
C10H11N
O2
263.14
05
[M+2TMS
+H]+
118.0536
118.062
9
9.3
C5H10O3
289.13
80
[M+2TMS
+H]+
144.0511
144.042
2
8.9
C6H8O4
175.0869
175.095
6
8.7
C6H13N3
O3
117.0636
117.053
8
9.8
153.0848
153.078
9
5.9
244.0616
244.069
5
190.12
89
442.25
07
[M+2TMS
+H]+
[M+3TMS
+H]+
[M+5TMS
+H]+
[M+3TMS
+H]+
[M+4TMS
+H]+
[M+5TMS
+H]+
[M+4TMS
+H]+
461.18
80
[M+3TMS
+H]+
320.17
38
334.19
00
C3H7N3O
2
7.9
S-8
C8H11NO
2
C9H12N2
O6
5Hydroxytryptoph
ol
3-Hydroxyvaleric
acid
3Methylglutaconic
acid
HMDB018
55b
HMDB005
31c
HMDB022
66d
Citrulline
HMDB009
04e
Guanidoacetic
acid
HMDB001
28
Dopamine
HMDB000
73f
Uridine
HMDB002
96g
461.18
80
[M+4TMS
+H]+
172.0220
172.013
6
8.4
468.23
95
[M+4TMS
+H]+
179.0735
179.079
3
5.8
492.26
57
275.1392
275.148
1
8.8
439.2299
439.239
2
513.2819
526.25
93
[M+3TMS
+H]+
[M+TMS+
H]+
[M+2TMS
+H]+
[M+4TMS
+H]+
[M+H]+
[M+TMS+
H]+
[M+2TMS
+H]+
[M+4TMS
+H]+
700.34
62
[M+2TMS
+H]+
512.27
72
514.28
97
Glycerol 3phosphate
HMDB001
26h
Glucosamine
HMDB015
14i
C11H21N3
O5
Epsilon-(gammaGlutamyl)-lysine
HMDB038
69j
9.3
C23H37N
O5S
Leukotriene E4
HMDB022
00
513.276
0
5.9
C26H43N
O7S
Sulfolithocholylg
lycine
HMDB026
39
237.0934
237.086
1
7.2
C9H11N5
O3
Biopterin
HMDB00
468k
555.2593
555.269
2
9.9
C28H37N5
O7
Enkephalin L
HMDB010
45
C3H9O6P
C6H13NO
5
700.34 [M+6TMS
267.095
C9H17NO
HMDB008
267.1012
5.8
Neuraminic acid
62
+H]+
4
30
8
a
b
One isomer found with source ID HMDB12140; One isomer found with source ID HMDB12490; cTen
isomers found with source IDs HMDB02011, HMDB00351, HMDB00410, HMDB00642, HMDB01863,
HMDB00354, HMDB00396, HMDB00407, HMDB01987, HMDB00754; dThree isomers found with
source IDs HMDB13311, HMDB02266, HMDB00393; eOne isomer found with source ID HMDB03148;
f
One isomer found with source ID HMDB04825; gOne isomer found with source ID HMDB00767; hOne
isomer with source ID HMDB02520; iOne isomer found with source ID HMDB02030; jOne isomer found
with source ID HMDB04207; kFive isomers found with source IDs HMDB01195, HMDB02263,
HMDB00633, HMDB00817, HMDB00238.
S-9