Transcript Figure 1. Metabolomics platform for the characterization of medicinal plants integrating GC/MS, LC/MS, alignment tools and statistical analysis using COVAIN (Sun.

Figure 1. Metabolomics platform for the characterization of medicinal plants integrating GC/MS, LC/MS, alignment tools and statistical analysis using
COVAIN (Sun & Weckwerth, 2012)
Gas chromatography coupled to
mass spectrometry
Liquid chromatography coupled to
mass spectrometry
MetMAX – Data alignment
COVAIN – Data integration,
Statistics and Modelling
Data mining
Figure 2. HPLC/ESI(OrbiTrap)/MS profile (full msmode) of the crude hydroalcoholicextract of P. anserina (positive ion mode)
7
9
15
10
16
12
6
8
13
3
11
1
2
4
5
14
17
Figure 3A. Sample patterns of the hydroalcoholic extracts of P.anserinaanalyzed by GC/MS.
P. anserina UniVie
1
5
3
4
2
P. anserina UniSa
Figure 3B.Scores plot of the hydroalcoholic extracts of P.anserinaanalyzed by LC/MS
(only identified compounds used as variables).
5
P. anserina
UniSa
3
1
P. anserina UniVie
4
2
Figure 3C. Sample patterns of the hydroalcoholic extracts of P.anserina analyzed by LC/MS
(all the compounds with RSD<25 used as variables).
1
P. anserina UniVie
5
P. anserina UniSa
2
4
3
Figure 4. Sample patterns of combined GC/MS and LC/MS data of the hydroalcoholic extracts of P.anserina.
1
2
3
4
5
PanserinaUniSa
PanserinaUniVie
4
3
PC 3 (12%)
2
1
0
-1
-2
-3
6
4
6
2
4
0
2
-2
0
-2
-4
-4
-6
-6
-8
PC 2 (24%)
-8
PC 1 (47.5%)
Intensity/total counts
Figure 5. Relative quantitative analysis of genistein in the commercial products (1, 2, 3, 4, 5) and in the two
hydroalcoholic crude extracts of P. anserina.
Supplementary data
Supplementary Figure 1.nanoLC/ESI(Orbitrap)/MS profile (full ms mode) of the crude hydroalcoholic extract of P. anserine and its commercial products (positive
Supplementary Figure 2.GC/EI(TSQ)/MS profile (full ms mode) of the crude hydroalcoholic extract of P. anserina and its commercial products (positive ion mode
Supplementary Figure 3. Loadings plot of the hydroalcoholic extracts of P.anserina analyzed by GC/MS.
Supplementary Figure 4. Loadings plot of the hydroalcoholic extracts of P.anserina analyzed by LC/MS (identified compounds).
Supplementary Figure 5 .Rt, LC/MS and LC/MSMS spectra comparison between reference standards and P. anserinahydroalcoholic
extract compounds.A:chlorogenic acid, [M+Na]+: 377; B: genistein, [M+H]+: 271; C: quercetin 3-O-glucoside, [M+H]+: 465; D: kaempferol
3-O-glucoside, [M+H]+: 449; E: myricetin 3-O-rhamnoside [M+H]+: 465; F: quercetin 3-O-glucuronide [M+H]+: 479
RT : 0.00 - 100.01
SM : 11G
RT : 0.00 - 44.91
50
4.97
57.64
33.90
43.32
355.10
465.10
303.05
371.10
387.18
81.65
99.77
371.10
371.10
391.28
19.02
50
12.75
371.10
668.44
m /z= 50.00-2000.00 F: IT M S + c NSI
371.10
[150.00-1000.00] M S
panseri nauni sa120ngul posddep060412
NL: 3.22E3
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
100
Ful l m s2 377.05@ci d40.00 [90.00-390.00] M S
Relative Abundance
Relative Abundance
d Ful l m s2 377.08@ci d40.00
[90.00-390.00] M S M i xST D1_060412
50
0
19.10
NL: 6.03E6
377.08
panseri nauni sa120ngul posddep060412
50
0
25.59
19.58
m /z= 376.50-377.50 F: FT M S + p NSI
100
100
M i xST D1_060412
4.77
23.14
17.09
376.95
37.43
377.08
377.09
43.28
377.02
54.54
377.02
72.61
68.57
377.10
78.87
377.33 377.34
0.00
50
84.67
6.03
0.96
377.17
m /z= 376.50-377.50 F: FT M S + p NSI Ful l m s
panseri nauni sa120ngul posddep060412
377.08
377.08
0.00
[150.00-1000.00] M S
21.19
14.05
10.31
377.16
0.00
0
NL: 1.18E6
377.05
377.08
Ful l m s [150.00-1000.00] M S
50
Base Peak F: FT M S + p NSI Ful l m s
641.14 163.04
0
NL: 8.88E3
215.07
7.64
294.15
37.66
493.10
465.10
25.59
22.49
NL: 1.41E7
44.86
33.48
6.11
M i xST D1_060412
76.01
100
A
100
[150.00-1000.00] M S
158.00
0
34.43
479.08
Base Peak F: FT M S + p NSI Ful l m s
70.65
19.07
SM : 11G
NL: 1.94E8
46.71
271.06
100
29.85
34.80
43.73
377.07
377.12
377.29
30
35
0
0
10
20
30
40
50
60
70
80
90
100
0
5
10
15
20
T i m e (m i n)
M i xST D1_060412 #1754
RT : 19.02
AV: 1
25
40
T i m e (m i n)
NL: 1.34E4
panseri nauni sa120ngul posddep060412 #1694
F: IT M S + c NSI d Ful l m s2 377.08@ci d40.00 [90.00-390.00]
RT : 19.17
AV: 1
NL: 1.45E2
F: IT M S + c NSI d Ful l m s2 377.05@ci d40.00 [90.00-390.00]
215.07
359.11
100
100
90
90
80
80
70
70
60
60
Relative Abundance
Relative Abundance
359.07
50
40
30
185.07
197.09
20
215.11
50
40
361.10
30
331.14
331.07
20
197.21
179.08
297.07
179.04
10
10
163.09
151.13
117.05
216.91
258.17
299.02 315.14
271.14
0
E:\DATA\ORBI\ORBISTDMIX\MixSTD1_060412
100
120
140
160
180
200
341.10
220
240
260
280
300
320
340
216.77
137.17
115.11
360.02
247.06
177.72
360
380
100
120
140
160
180
200
220
240
m /z
RT : 0.00 - 100.01
RT : 39.48 - 50.00
271.06
33.90
43.32
465.10
303.05
668.44
100
M i xST D1_060412
76.01
81.65
99.77
371.10
371.10
391.28
153.00
m /z= 50.00-2000.00 F: IT M S + c NSI
100
Relative Abundance
Relative Abundance
46.71
NL: 2.05E8
271.06
panseri nauni sa120ngul posddep060412
49.25
527.33
43.38
NL: 7.71E2
137.03
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
100
panseri nauni sa120ngul posddep060412
50
0
m /z= 270.50-271.50 F: FT M S + p NSI
100
48.03
NL: 4.55E5
271.19
m /z= 270.50-271.50 F: FT M S + p NSI Ful l m s
100
Ful l m s [150.00-1000.00] M S
B
380
Ful l m s2 271.06@ci d40.00 [60.00-285.00] M S
[60.00-285.00] M S M i xST D1_060412
215.06
[150.00-1000.00] M S
43.36
M i xST D1_060412
50
360
[150.00-1000.00] M S
47.54
266.17
d Ful l m s2 271.06@ci d40.00
69.34
0
368.14
340
Base Peak F: FT M S + p NSI Ful l m s
47.16
371.10
595.14
0
NL: 8.71E4
45.71
43.57
593.19
41.36
50
797.25
46.88
50
341.10
320
NL: 1.26E7
668.44
41.77
[150.00-1000.00] M S
371.10
387.18
158.00
0
300
44.86
Base Peak F: FT M S + p NSI Ful l m s
70.65
57.64
19.07
355.10
280
SM : 11G
NL: 1.94E8
100
4.97
260
m /z
SM : 11G
46.71
50
315.12
279.01
260.99
0
06/04/2012 14:40:58
1.95
11.92
27.23
32.45
43.94
47.96
63.29
67.54
73.53
82.32
97.28
271.19
271.19
271.19
271.08
271.15
271.06
271.06
271.06
271.06
271.06
271.19
39.90
41.09
42.04
271.06
271.11
270.98
50
0
46.95
44.56
46.22
271.10
271.06
271.10
panseri nauni sa120ngul posddep060412
49.93
271.06
271.23
0
0
10
20
30
40
50
60
70
80
90
100
40
41
42
43
44
T i m e (m i n)
M i xST D1_060412 #4130
RT : 45.86
AV: 1
45
46
47
48
49
50
T i m e (m i n)
NL: 3.39E2
panseri nauni sa120ngul posddep060412 #4112
F: IT M S + c NSI d Ful l m s2 271.06@ci d40.00 [60.00-285.00]
RT : 46.91
AV: 1
NL: 2.69E2
F: IT M S + c NSI d Ful l m s2 271.06@ci d40.00 [60.00-285.00]
253.05
100
153.08
282.76
153.05
100
90
90
215.06
80
253.08
80
282.72
70
70
243.16
50
40
149.05
163.19
30
107.14
20
243.12
50
40
30
224.98
173.13
145.11
149.11
20
121.17
81.04
187.12
93.11
10
215.05
60
Relative Abundance
Relative Abundance
60
121.07
254.26
272.95
10
85.11
0
93.17
185.04
107.02
80
100
120
140
160
180
panserinaunisa120ngulposddep060412
RT : 0.00 - 50.31
200
220
240
260
280
60
80
100
211.20
140
160
RT : 0.00 - 44.91
46.71
33.90
19.07
4.97
158.00
9.73
371.10
12.02
25.01
355.10
686.32
371.10
30.18
465.10
832.43
0
36.59
43.32
449.11
303.05
34.43
Base Peak F: FT M S + p NSI Ful l m s
303.08
294.15
7.64
12.75
22.49
25.59
371.10
371.10
641.14
163.04
668.44
493.10
Relative Abundance
[115.00-480.00] M S
M i xST D1_060412
0
33.92
NL: 4.20E7
465.10
4.95
10.32
16.14
19.90
22.34
29.69
464.92
465.10
465.10
465.07
465.10
465.10
35.92
39.46
46.85
465.10
465.10
465.10
panseri nauni sa120ngul posddep060412
33.48
NL: 7.84E6
465.10
m /z= 464.50-465.50 F: FT M S + p NSI Ful l m s
100
[150.00-1000.00] M S
34.41
50
0
Base Peak F: FT M S + p NSI Ful l m s
Ful l m s2 465.10@ci d40.00 [115.00-480.00] M S
0
M i xST D1_060412
33.15
465.10
NL: 1.41E7
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
Ful l m s [150.00-1000.00] M S
50
280
NL: 7.40E3
303.02
100
50
m /z= 464.50-465.50 F: FT M S + p NSI
100
260
panseri nauni sa120ngul posddep060412
35.87
d Ful l m s2 465.10@ci d40.00
50
240
[150.00-1000.00] M S
465.10
0
m /z= 50.00-2000.00 F: IT M S + c NSI
37.66
33.48
50
NL: 1.34E5
100
220
44.86
479.08
100
6.11
M i xST D1_060412
34.35
200
SM : 11G
NL: 1.94E8
[150.00-1000.00] M S
50
180
m /z
SM : 11G
271.06
5.99
9.80
15.91
20.26
22.49
27.66
31.52
465.21
465.07
465.07
465.10
465.10
465.14
465.10
465.10
panseri nauni sa120ngul posddep060412
39.29
465.10
0
0
5
10
15
20
25
30
35
40
45
50
0
5
10
15
T i m e (m i n)
M i xST D1_060412 #3112
RT : 34.35
AV: 1
20
25
30
35
40
T i m e (m i n)
NL: 5.59E5
panseri nauni sa120ngul posddep060412 #3000
F: IT M S + c NSI d Ful l m s2 465.10@ci d40.00 [115.00-480.00]
RT : 34.33
AV: 1
NL: 2.30E4
F: IT M S + c NSI d Ful l m s2 465.10@ci d40.00 [115.00-480.00]
303.08
303.04
100
100
90
90
80
80
70
70
60
60
Relative Abundance
Relative Abundance
272.94
271.12
120
07/04/2012 05:58:27
m /z
100
Relative Abundance
197.21
0
60
C
254.70
229.08
163.22
145.04
229.29
197.05
80.26
50
40
30
50
40
30
20
20
10
10
144.92
162.96
229.05
247.36
285.14
315.04
345.07
369.11
399.13
429.08
447.07
0
147.09
163.20
199.17
237.36
277.08
289.16
319.10
345.14
369.14
399.11
429.18
447.21
0
150
200
250
300
m /z
350
400
450
150
200
250
300
m /z
350
400
450
RT : 29.46 - 41.82
SM : 11G
RT : 24.96 - 41.99
34.29
31.50
29.64
451.09
371.10
33.56
32.14
371.10
38.12
493.10
509.09
39.84
517.13
50
40.55
25.59
163.04
371.10 371.10
0
NL: 1.41E7
26.37
29.67
595.17
565.16
Base Peak F: FT M S + p NSI Ful l m s
493.10
36.78
465.10
41.77
38.58
287.05
495.08
34.35
NL: 1.10E4
287.03
Base Peak m /z= 50.00-2000.00 F: IT M S +
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
100
[110.00-460.00] M S M i xST D1_060412
0
36.59
NL: 2.38E7
449.11
0
31.76
33.15
449.11 449.11 449.11
34.87
449.11
38.39
35.76
449.11
39.49
449.11
449.11
25.20
[150.00-1000.00] M S
449.11
29.18
449.11
449.11
0
449.11
30.42
33.08
34.41
449.11
449.07
449.11
panseri nauni sa120ngul posddep060412
38.87
39.70
449.11 449.11
0
30
31
32
33
34
35
36
37
38
39
40
41
26
28
30
32
34
T i m e (m i n)
M i xST D1_060412 #3242
RT : 35.77
AV: 1
36
38
40
T i m e (m i n)
NL: 9.48E3
panseri nauni sa120ngul posddep060412 #3136
F: IT M S + c NSI d Ful l m s2 449.11@ci d40.00 [110.00-460.00]
RT : 35.85
AV: 1
NL: 3.60E3
F: IT M S + c NSI d Ful l m s2 449.07@ci d40.00 [110.00-460.00]
287.06
287.04
100
100
90
90
80
80
70
70
60
60
Relative Abundance
Relative Abundance
m /z= 448.50-449.50 F: FT M S + p NSI Ful l m s
29.85
50
41.34
449.11
NL: 4.95E6
449.11
100
M i xST D1_060412
31.04
36.80
m s [150.00-1000.00] M S
449.11
29.55
panseri nauni sa120ngul posddep060412
50
m /z= 448.50-449.50 F: FT M S + p NSI Ful l
37.43
100
50
Ful l m s2 449.07@ci d40.00 [110.00-460.00] M S
Relative Abundance
Relative Abundance
c NSI d Ful l m s2 449.11@ci d40.00
50
[150.00-1000.00] M S
593.19 panseri nauni sa120ngul posddep060412
648.41
0
NL: 7.92E3
37.66
33.48
30.65
37.56
100
D
100
[150.00-1000.00] M S m i xstd2_060412
34.82
451.09
34.43
479.08
Base Peak F: FT M S + p NSI Ful l m s
36.83
449.11
50
SM : 11G
NL: 3.03E7
479.08
100
50
40
30
50
40
30
20
20
10
153.89
185.30
223.32 238.90
panserinaunisa120ngulposddep060412
0
150
200
303.05
278.99
250
329.07 345.01
300
383.14
419.11
07/04/2012
05:58:27
350
433.10
10
433.13
147.10
177.68
205.27
227.11
256.43
450
150
200
250
300
m /z
RT : 30.72 - 37.03
RT : 30.66 - 36.18
33.46
34.43
31.40
31.71
32.32
34.02
34.45
35.38
35.71
371.10
371.10
371.10
465.10
465.10
303.05
371.10
100
597.15
31.79
32.99
371.10
451.09
35.34
303.05
panseri nauni sa120ngul posddep060412
35.86
303.05
0
Base Peak m /z= 49.50-50.50 F: IT M S + c
100
[150.00-1000.00] M S
465.10
31.52
50
NL: 0
NL: 7.40E3
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
100
Ful l m s2 465.10@ci d40.00 [115.00-480.00] M S
Relative Abundance
Relative Abundance
NSI d Ful l m s2 465.10@ci d40.00
[115.00-480.00] M S m i xstd3_060412
50
33.46
NL: 1.54E7
465.10
panseri nauni sa120ngul posddep060412
50
0
33.48
NL: 7.84E6
465.10
m /z= 464.50-465.50 F: FT M S + p NSI
100
m /z= 464.50-465.50 F: FT M S + p NSI Ful l m s
100
Ful l m s [150.00-1000.00] M S
E
31.56
34.02
32.41
465.10 465.10
465.10
465.10
450
Base Peak F: FT M S + p NSI Ful l m s
33.48
[150.00-1000.00] M S m i xstd3_060412
435.09
0
31.16
440.40
400
NL: 1.41E7
479.08
Base Peak F: FT M S + p NSI Ful l m s
36.66
50
403.18
SM : 11G
NL: 1.48E7
100
0
383.18
350
m /z
SM : 11G
465.10
50
329.17 345.06
315.08
285.14
0
400
[150.00-1000.00] M S
m i xstd3_060412
34.45
35.49
465.10
35.90
50
36.18
31.52
31.13
32.44
465.10 465.10
465.10 465.10 465.10
0
33.98
32.90
465.10
465.10 465.10
34.41
panseri nauni sa120ngul posddep060412
35.86
465.10
465.10
0
31
32
33
34
35
36
37
31
32
33
T i m e (m i n)
m i xstd3_060412 #2804
RT : 33.10
AV: 1
34
35
36
T i m e (m i n)
NL: 2.44E3
panseri nauni sa120ngul posddep060412 #2910
F: IT M S + c NSI d Ful l m s2 465.10@ci d40.00 [115.00-480.00]
RT : 33.31
AV: 1
NL: 2.69E3
F: IT M S + c NSI d Ful l m s2 465.10@ci d40.00 [115.00-480.00]
319.01
319.04
100
100
90
90
80
80
70
70
60
60
Relative Abundance
Relative Abundance
303.06
50
40
30
50
40
30
20
20
10
178.08
236.98
e:\data\orbi\orbistdmix\mixstd3_060412
0
150
200
303.06
262.21
250
329.01
300
429.02
361.10
411.13
06/04/2012 22:19:43
350
447.16
10
447.29
459.99
147.07
165.91
208.93
240.29 259.15
450
150
200
250
300
m /z
RT : 30.72 - 37.03
SM : 11G
RT : 30.66 - 36.18
33.46
31.40
31.71
32.32
34.02
34.45
35.38
35.71
371.10
371.10
371.10
465.10
465.10
303.05
371.10
435.09
449.99
450
Base Peak F: FT M S + p NSI Ful l m s
33.48
[150.00-1000.00] M S
465.10
31.52
50
31.79
32.99
371.10
451.09
597.15
35.34
303.05
35.86
Base Peak m /z= 49.50-50.50 F: IT M S + c
NL: 6.26E3
Base Peak m /z= 50.00-2000.00 F: IT M S + c NSI d
100
Relative Abundance
NSI d Ful l m s2 479.08@ci d40.00
[120.00-490.00] M S m i xstd3_060412
50
0
NL: 7.13E5
34.47
479.08
Ful l m s2 479.08@ci d40.00 [120.00-490.00] M S
panseri nauni sa120ngul posddep060412
50
0
m /z= 478.50-479.50 F: FT M S + p NSI
100
34.43
NL: 1.46E7
479.08
m /z= 478.50-479.50 F: FT M S + p NSI Ful l m s
100
Ful l m s [150.00-1000.00] M S
m i xstd3_060412
30.83
31.68
32.48
33.23
35.33
36.09
36.66
479.08
479.08
479.12
479.08
479.08
479.08
479.06
[150.00-1000.00] M S
50
0
panseri nauni sa120ngul posddep060412
303.05
0
NL: 0
100
panseri nauni sa120ngul posddep060412
31.10
31.65
32.39
33.01
34.00
35.50
479.08
479.08
479.15
479.15
479.12
479.08
0
31
32
33
34
35
36
37
31
32
33
T i m e (m i n)
m i xstd3_060412 #2910
RT : 34.34
AV: 1
34
35
36
T i m e (m i n)
NL: 4.00E3
panseri nauni sa120ngul posddep060412 #2982
F: IT M S + c NSI d Ful l m s2 479.08@ci d40.00 [120.00-490.00]
RT : 34.13
AV: 1
NL: 5.98E3
F: IT M S + c NSI d Ful l m s2 479.08@ci d40.00 [120.00-490.00]
303.06
303.06
100
100
90
90
80
80
70
70
60
60
Relative Abundance
Relative Abundance
428.91
400
NL: 1.41E7
479.08
100
[150.00-1000.00] M S m i xstd3_060412
0
Relative Abundance
34.43
Base Peak F: FT M S + p NSI Ful l m s
36.66
F
403.12
SM : 11G
NL: 1.48E7
465.10
50
361.11
350
m /z
100
50
329.07
285.09
0
400
50
40
30
50
40
30
20
20
10
10
158.56
217.29
241.21 262.80 275.01
317.00
345.10
380.91
398.90
443.25
461.01
0
317.07
147.12
163.05 183.20
239.11
256.93
299.16
347.01 369.16
399.04
424.91
438.03
461.08
0
150
200
250
300
m /z
350
400
450
150
200
250
300
m /z
350
400
450