Supporting Information
Table S1. Elemental compositions of product ions of metabolites, M1–M19
Metabolites
M1-M19
Formula
Observed mass (Da)
Calculated mass (Da)
Error (ppm)
C9H8NO4
C9H8NO3
C7H8NO4
C14H14FN2O6
C14H11FNO6
194.0441
178.0491
170.0451
325.0835
308.0560
194.0448
178.0499
170.0448
325.0830
308.0565
3.35
3.39
-1.05
-1.51
1.50
C12H12NO6
266.0651
266.0659
3.04
C11H9FNO3
222.0559
222.0561
0.76
C10H17N2O2
197.1281
197.1285
1.15
C9H14FN2O
185.1088
185.1085
-0.93
C7H14FN2O
C9H15N2O
161.1081
167.1171
161.1085
167.1179
1.13
3.01
C17H16FN2O6
363.0981
363.0987
1.66
C13H12FN2O6
311.0672
311.0674
0.65
C13H9FNO6
C11H10NO6
294.0413
252.0509
294.0408
252.0503
-1.44
-1.62
C18H23FN3O2
C17H18FN2O2
C9H14FN2O3S
332.1773
301.1343
249. 0710
332.1769
301.1347
249.0704
-1.15
1.11
-1.30
C13H11FNO2
232.0769
232.0768
-0.07
C20H23FN3O2
C17H18FN2O3
C13H14FN2O3
C13H11FNO3
356.1774
317.1299
265.0988
248.0713
356.1769
317.1296
265.0983
248.0717
-1.03
-0.90
-1.35
1.13
C13H9FNO2
230.0616
230.0612
-1.16
C6H8NO2
C19H23FN3O2
C21H27FN3O2
C11H19N2O
C10H16FN2
C8H16FN2
C12H19N2O2
C11H16FN2O
C9H16FN2O
C10H17N2O4S
C9H14FN2O3S
C7H14FN2O3S
C6H9O6
126.0561
344.1761
372.2087
195.1495
183.1296
159.1295
223.1446
211.1238
187.1248
261.0907
249.0709
225.0708
177.0391
126.0550
344.1769
372.2082
195.1492
183.1292
159.1292
223.1441
211.1241
187.1241
261.0904
249.0704
225.0704
177.0394
-3.98
2.01
-1.33
-0.91
-1.12
-0.91
-1.34
0.95
-1.95
-0.92
-1.32
-1.15
0.92
1
N
H
H
F
OH
N
H
m/ z 145.1131
N
N
H
H
1
R
O
OH
F
O
1
M4
H
OH
R
R
1
m/ z 318.1619 R = H M4 & M5
m/ z 334.1563 R1= OH M6
OH
F
1
R
O
R1= H
- C7H 14N2
N
H
- C3H7N
R=H
N
N
N
m/ z 262.0526
N
N
H
- C3H2O3 M5 & M6
m/ z 404.1611 R=OH R1= H M5
m/ z 420.1571 R=OH R1= OH M6
F
H
OH
M4
O
R=H R1= H
m/ z 388.1661
N
H
- C3 H2O2
OH
F
N
m/ z 169.1131
R
N
m/ z 167.1171
OH
H
O
OH
M4
N
N
R
N
1
R=OH R = H M5
m/ z 278.0452
1
m/ z 294.0413
R=OH R = OH
F
M6
OH
F
1
R
O
O
m/ z 331.1093
R=H R = H
m/ z 347.1033
R=OH R 1= H M5
R
M4
m/ z 363.0981 R=OH R1= OH M6
- C4 H4
O
1
1
m/ z 192.0458
R1= H
m/ z 208.0411
R = OH M6
M4
1
H
OH
H2N
N
R
OH
F
1
R
O
O
R1= H
m/ z 279.0772
R=H
m/ z 295.0722
R=OH R1= H
M4
m/ z 311.0672
1
R=OH R = OH M6
M5
- C2H2NF
H
H
N
R
OH
- C4H2O
M4 & M5
- C4H2 O2 M6
HO
N
R
O
O
m/ z 154.0493
R
R=H M4
m/ z 170.0451 R=OH M5 & M6
OH
1
O
m/ z 220.0611
- C2H2 O M4 & M5
HC
N
R
M4
OH - C3H2 O3 M5 & M6
- C2H2O2 M6
O
- C 3H2O2
O
C6H6NO
+
m/ z 108.0449
M4, M5 & M6
O
1
R=H R = H M4
m/ z 236.0558 R=OH R 1= H M5
m/ z 252.0509 R=OH R1= OH M6
m/ z 178.0491
R=H M4
m/ z 194.0441
R=OH M5 & M6
Scheme S1. Proposed fragmentation mechanism for protonated metabolites M4 (m/z 388), M5
(m/z 404) and M6 (m/z 420).
2
H
H
OCH3
OCH3
N
N
N
H
N
- C2H2 M7
N
H
m/ z 181.1338
F
- C2H2O
R
M8
H
N
H
m/ z 169.1131
m/ z 374.1880
F
- H2O
R= H M7
m/ z 358.1922
N
N
H
O
N
O
m/ z 332.1773
M8
R= OH M8
H
OCH3
N
- NH3
F
m/ z 356.1174
- C3H7N
H
H
N
F
m/ z 341.1660
OCH3
N
H
M7
N
N
F
R
O
m/ z 301.1343
m/ z 145.1131
- C 4H4
m/ z 317.1299
R= H
H
m/ z 230.0616
H
OCH3
OCH 3
H 2N
N
- H2O
M7
R= OH M8
N
N
- NH3
M8
F
R
m/ z 232.0769
O
R= H
m/ z 248.0713
R= OH M8
M7
F
R
O
m/ z 249.0710
R= H
m/ z 265.0988
R= OH M8
M7
O
- H2O
R
m/ z 110.0611 R= H
m/ z 126.0561
m/ z 108.0449
M8
M7
R= OH M8
Scheme S2. Proposed fragmentation mechanism for protonated metabolites M7 (m/z 358) and
M8 (m/z 374).
3
H
HO
H
H
OH
O
N
N
- CO2
N
m/ z 167.1171
N
N
H
OH
OH
F
H
F
N
OH
N
H
N
H
OH
F
O
O
O
O
m/ z 388.1664
m/ z 432.1560
- 2 CO2
N
N
H
- CO2
- C3H7N
m/ z 169.1131
N
N
H
H
H
F
N
OH
H
O
N
m/ z 145.1131
OH
N
OH
F
OH
F
N
N
H
m/ z 344.1761
O
O
O
m/ z 375.0981
- C4H4
OH
O
N
H
OH
OH
- NH3
H2 N
O
N
OH
F
O
H
OH
OH
O
- C 6H4FNO
O
OH
O
O
m/ z 323.0672
m/ z 306.0412
O
- C2H2FN
m/ z 134.0603
HO
N
- C2H2O
- 2 CO2
H
O
N
OH
- 2 CO2
OH
O
m/ z 176.0709
m/ z 198.0393
O
H
H
HO
OH
N
F
O
O
m/ z 264.0509
N
O
m/ z 110.0605
Scheme S3. Proposed fragmentation mechanism for protonated metabolite M9 (m/z 432).
4
OCH3
H
N
H
N
R
m/ z 195.1495
N
R= -CH3
M10
R= SO3H
R
N
m/ z 444.1936
R= COCH 3 M11
R= -CH 3
m/ z 482.1397
R= SO3H
- SO3
OH
F
m/ z 402.1821
M10
O
O
- CO2
M12
H
F
M10
- C4H9 N
M10
OCH3
N
- C5H 9NO M11
m/ z 211.1238 R= COCH3 M11
m/ z 249.0709
R= SO3H
N
N
H
O
m/ z 416.1971
N
N
- CH2 CO
F
O
R= -CH3
H
m/ z 183.1296
M11 & M12
N
OH
M12
R
OCH 3
N
m/ z 223.1446 R= COCH3 M11
m/ z 261.0907
H
OCH3
- C3H7NO3S M12
M12
N
F
H
N
R= -CH 3
R= COCH3 M11
m/ z 225.0708
R= SO3 H
m/ z 372.2087 R= -CH3
F
- CH2 CO M11
- SO3
M12
N
OH
F
M10
O
OCH 3
R
m/ z 159.1295
R
H
N
m/ z 187.1248
N
N
N
M10, M11 & M12
m/ z 438.1496
R= SO3H
M12
OCH3
m/ z 345.1248
M12
R= COCH3 M11
H
O
O
M10
m/ z 400.2035
N
N
H
F
- C4H4
O
m/ z 358.1922
M11 & M12
H
H
OCH3
H2N
H
N
- CO2
N
OH
F
O
O
O
m/ z 154.0493
O
m/ z 293.0937
N
OH
O
m/ z 110.0611
Scheme S4. Proposed fragmentation mechanism for protonated metabolites M10 (m/z 416), M11
(m/z 444) and M12 (m/z 482).
5
H
O
C
OH
N
O
OH
OH
N
O
OH
OH
N
O
N
N
H
OH
O
m/ z 507.1403
O
H
OH
O
N
- 176 Da
H
OH
N
N
N
N
H
OH
O
m/ z 388.1661
m/ z 262.0526
N
N
N
H
F
O
O
H
OH
- C3H2O2
F
O
N
H
m/ z 167.1171
OH
OH
OH
HO
O
OH H
- C6H8O6
OH m/ z 177.0391
F
OH
m/ z 438.0837
OH
OH
HO
O
F
m/ z 564.1980
C
O
N
OH
O
O
C OH
OH
OH
- C17H14N2
O
F
O
O
O
C OH
- C3H7N
O
F
H
O
m/ z 318.1619
- C3H7N
- C7 H14N2
H
OH
OH
N
N
N
OH
F
O
O
m/ z 331.1093
F
O
m/ z 192.0458
- C4 H4
H
OH
H2N
N
OH
F
O
O
m/ z 279.0772
- C2H2NF
H
H
- C 4H2O
N
HO
N
OH
O
O
m/ z 154.0493
OH
O
O
m/ z 220.0611
Scheme S5. Proposed fragmentation mechanism for protonated metabolite M15 (m/z 564).
6
H
H
O
OCH3
N
C OH
N
O
OCH3
- C 3H 7N
N
OH
O
F
m/ z 493.1612
O
O
OH
O
F
OH
O
N
N
H
OH
O
OCH3
C OH
N
- C7H14N2
OH
O
OH
O
F
OH
m/ z 550.2189 O
C OH
OH
OH
O
m/ z 424.1032
H
O
O
HO
- C6H8O6
C
OH
- 176 Da
OH
OH m/ z 177.0391
OCH3
H
N
N
H
F
N
m/z 169.1131
H
N
N
H
OH
m/ z 374.1880
N
H
OCH3
N
- C2H2O
O
m/ z 181.1338
N
- H2O
- C3H7N
F
N
N
H
F
m/ z 356.1174
H
m/ z 145.1131
O
m/z 332.1773
H
N F
N
H
H
OCH3
H
OCH3
N
N
F
OH
O
m/ z 317.1299
- C 4H 4
H
H
OCH 3
H2N
N
F
N
OH
O
m/ z 265.0988
O
- H 2O
m/ z 108.0449
OH
m/ z 126.0561
Scheme S6. Proposed fragmentation mechanism for protonated metabolite M16 (m/z 550).
7
OCH 3
OH
- C 3H7N
N
OH
O
O
C
O
F
O
O
HO
OH
OH
O
OH
HO
H
OH
m/z 644.1252
C
OH
O
O
O
OH
- 1 76 Da
O
O
- C6H 8O6
O
O
H
O
H
O CH 3
N
C
N
OH
OCH 3
OH
- C 3H7N
N
O
OH
O
O
O
OH
O
O
OH
HO
OH
HO
OH
- C7H 14N2
m/ z 537.1518
H
C
O
OH
O
m/ z 594.2091
OH
OH
m/ z 263.0392
O
F
O
F
OH
HO
C OH
OH
N
N
H
OH
O
C
OH
C
OH
OH
m/z 439.0714
HO
O
O
OH
O
H
OH
OH
- C6H 8O6
C
OH
O
O
O
O
F
OH
OH
OH
- C7H 14N 2
C
OH
O
m/z 770.2419
OH
HO
m/z 713.1838
O
O
OH
O
N
OH
O
OH
O
OCH 3
OH
O
N
N
H
O
F
O
O
O
OCH3
OH
OH
H
OH
O
C
C
C
N
OH
H
OH
OH
N
O
O
O
m/ z 1 77.03 91
- C 6H 8O6
O
OCH3
C
N
- 1 76 Da
OH
OH
O
OH
O
F
H
OH
O
O
N
m/ z 4 68.09 31
OCH 3
- C 3H2O3
N
OH
N
N
H
OH
F
N
N
H
O
F
m/ z 41 8.17 80 O
OCH 3
H
OCH 3
OH
O
m/ z 3 32.17 73
H
H
N
N
H
H
m/z 181.1338
N
NF
N
H
m/ z 145.1131
N
H
F
m/z 169.1131
Scheme S7. Proposed fragmentation mechanism for protonated metabolite M17 (m/z 770).
8
HO
HO
O C
HO
OH
H
O
OH
O
C
O
OH
O
N
O
F
N
OH
C
C
O
F
HO
O
- 176 Da
OH
C
O
F
O
N
OH
O
O
N
OH
O
N
N
H
OH
F
O
m/ z 343 .1506
OH
O
O
m/ z 438 .0837
N
H
OH
O
H
OH
OH
OH
- C 7H14N 2
O
N
OH
C
H
O
O
OH
m/ z 564 .1980
- C 3H 7N
H
O
- C 6H8O6
H
C
O
OH
N
O
N
OH
H
OH
HO
OH
OH
- 176 Da
C
O
OH
OH
N
OH
O
F
N
O
N
H
m/ z 388 .1661
O
OH
O
m/ z 507 .1411
OH
F
m/ z 177.0391
OH
m/ z 614 .1157
HO
- C 6H8O6
OH
OH
O
O
m/ z 683 .1733
C
OH
O
N
- C 7H14N 2
O
O
OH
O
OH
m/ z 740 .2316
O
O
OH
O
F
O
O
O
N
N
H
OH
O
C OH
OH
O
- C 3H 7N
OH
H
O
OH
O
O
N
HO
C
OH
OH
HO
OH
HO
O
H
N
H
NF
N
H
OH
N
H
m/ z 145.1131
F
m/ z 169.1131
N
OH
F
O
O
m/ z 262 .0526
Scheme S8. Proposed fragmentation mechanism for protonated metabolite M18 (m/z 740).
9
O
O
OH
C
OH
O
N
O
N
C
HO
O
O
F
OH
N
O
C
O
OH
N
OH
O
F
HO
O
C
OH
HO
OH
OH
HO
m/ z 263.0392
OH
O
C
O
OH
O
O
O
HO
O
OH
H
C
OH
HO
OH
HO
OH
- C 6H 8O6
- 17 6 Da
O
O
O
m/ z 580.1931
OH
F
O
C
O
OH
- C7H 14N2
OH
N
N
F
OH
O
O
HO
N
H
HO
H
O
m/ z 439.0714
N
OH
O
O
HO
m/ z 523.1352
O
OH
C
OH
OH
O
O
O
OH
H
OH
H
OH
OH
HO
- C 6H 8O6
O
O
N
C
HO
O
- C3H 7N
O
O
m/ z 630.1107
O
- C 6H 8O6
- 17 6 Da
H
O
OH
m/ z 167.1171
OH
OH
F
O
C
N
H
OH
O
H
OH
O
- C7H 14N2
HO
N
OH
OH
HO
HO
OH
O
OH
O
O
m/ z 756.2251
OH
H
OH
N
H
OH
m/ z 699.1683
OH
OH
N
O
O
OH
O
OH
O
O
C
- C3H 7N
O
F
H
OH
OH
N
C
O
m/ z 1 77.0 391
O
O
C
OH
HO
m/ z 454.0786
H
OH
HO
OH
N
N
H
OH
N
H
NF
N
H
m/ z 1 45.1 131
OH
F
m/ z 404.1611
O
O
- C3H 7N
OH
N
OH
OH
F
O
O
m/ z 278.0452
H
N
H
N
OH
N
OH
OH
F
O
N
H
F
m/ z 16 9.113 1
O
m/ z 347.1033
Scheme S9. Proposed fragmentation mechanism for protonated metabolite M19 (m/z 756).
10
Figure S1. (a) ESI-MS/MS spectrum of [M+H]+ ion (m/z 402) of MFX at 31 eV, (b) MS3
spectrum of m/z 358 ion at 29 eV, and (c) MS4 spectrum of m/z 301 ion at 33 eV.
11
Figure S2. LC/MS/MS spectra of (a) protonated M1 and (b) deuterated M1 at 28 eV.
12
Figure S3. LC/MS/MS spectra of (a) protonated M2 and (b) deuterated M2 at 28 eV.
13
Figure S4. LC/MS/MS spectra of (a) protonated M3 and (b) deuterated M3 at 28 eV.
14
Figure S5. LC/MS/MS spectra of (a) protonated M4 and (b) deuterated M4 at 25 eV.
15
Figure S6. LC/MS/MS spectra of (a) protonated M5 and (b) deuterated M5 at 25 eV.
16
Figure S7. LC/MS/MS spectra of (a) protonated M6 and (b) deuterated M6 at 25 eV.
17
Figure S8. LC/MS/MS spectra of (a) protonated M7 and (b) deuterated M7 at 31 eV.
18
Figure S9. LC/MS/MS spectra of (a) protonated M8 and (b) deuterated M8 at 31 eV.
19
Figure S10. LC/MS/MS spectra of (a) protonated M9 and (b) deuterated M9 at 31 eV.
20
Figure S11. LC/MS/MS spectra of (a) protonated M10 and (b) deuterated M10 at 25 eV.
21
Figure S12. LC/MS/MS spectra of (a) protonated M11 and (b) deuterated M11 at 25 eV.
22
Figure S13. LC/MS/MS spectra of (a) protonated M12 and (b) deuterated M12 at 25 eV.
23
Figure S14. LC/MS/MS spectra of (a) protonated M13 and (b) deuterated M13 at 25 eV.
24
Figure S15. LC/MS/MS spectra of (a) protonated M14 at 25 eV, (b) deuterated M14 at 25 eV
and, (c) MS3 spectrum of m/z 353 ion of protonated M14 at 28 eV.
25
Figure S16. LC/MS/MS spectra of (a) protonated M15 and (b) deuterated M15 at 25 eV.
26
Figure S17. LC/MS/MS spectra of (a) protonated M16 and (b) deuterated M16 at 25 eV.
27
Figure S18. LC/MS/MS spectra of (a) protonated M17 at 25 eV, (b) deuterated M17 at 25 eV
and, (c) MS3 spectrum of m/z 439 ion of protonated M17 at 28 eV.
28
Figure S19. LC/MS/MS spectra of (a) protonated M18 and (b) deuterated M18 at 25 eV.
29
Figure S20. LC/MS/MS spectra of (a) protonated M19 at 25 eV, (b) deuterated M19 at 25 eV
and, (c) MS3 spectrum of m/z 439 ion of protonated M19 at 28 eV.
30