DMD 28803

Supplemental materials

Legends for figures

Figure S1. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M2 (Rt = 8.35 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S1) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 239 observed in the product ion spectrum of M2 in panel A. Panel C depicts the MS3 spectrum of fragment ion m/z 322 observed in the product ion spectrum of M2 in panel A. The fragmentation pathways are similar to those of M1 (see Figure 7 and 8). The mass isolation width was set to 1 Da when obtained the above product ion spectra.

Figure S2. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M6 (Rt = 6.00 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S2) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M6 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

Figure S3. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M3 (Rt = 8.13 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S3) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M3 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

Figure S4. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M4 (Rt = 7.72 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S4) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M4 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

Figure S5. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M7 (Rt = 5.6 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S5) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M7 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

Figure S6. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M8 (Rt = 5.27 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S6) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M8 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

Figure S7. Product ion mass spectrum obtained by CID of the molecular ion of metabolite M9 (Rt = 4.60 min) at m/z 280.6 from NADPH-supplemented liver microsomal incubations containing nomifensine (10 mM) and GSH (10 mM) (see Table S7) (A). Panel B depicts the MS3 spectrum of fragment ion m/z 285 observed in the product ion spectrum of M9 in panel A. The origins of the major product ions are as indicated. The major fragmentation pathways are similar to those of M5 (Figure 10).

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Figure S2


Figure S3


Figure S4


Figure S5


Figure S6


Figure S7


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DMD 28803

Table S1. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M2 (Rt = 8.35 min) at m/z 280.6 (see Figure S1A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
323 / [C10H17O7N3S]+ / 323.0786 / 323.0782 / 1.3
322 / [C10H16O7N3S]+ / 322.0709 / 322.0704 / 1.7
287 / [C16H19ON2S]+ / 287.1218 / 287.1213 / 1.9
256 / [C10H14O5N3]+ / 256.0932 / 256.0928 / 1.6
247 / [C8H11O5N2S]+ / 247.0386 / 247.0383 / 1.1
239 / [C16H19N2]+ / 239.1546 / 239.1548 / 1.4
238 / [C16H18N2]+ / 238.1469 / 238.1464 / 1.9
223 / [C15H15N2]+ / 223.1233 / 223.1230 / 1.4
181 / [C8H9O3N2]+ / 181.0608 / 181.0608 / 0.17
145 / [C5H9O3N2]+ / 145.0607 / 145.0608 / -0.47

Table S2. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M6 (Rt = 6.00 min) at m/z 280.6 (see Figure S2A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1849 / 485.1853 / -0.86
431 / [ C21H27O4N4S ]+ / 431.1747 / 431.1748 / -0.12
422 / [ C23H24O3N3S ]+ / 422.1534 / 422.1533 / 0.26
328 / [ C18H22ON3S ]+ / 328.1480 / 328.1484 / 0.58
311 / [ C18H19ON2S ]+ / 311.1214 / 311.1218 / 0.48
287 / [C16H19ON2S]+ / 287.1215 / 287.1213 / 0.83
285 / [C16H17ON2S]+ / 285.1059 / 285.1062 / 1.0
249 / [C25H30O5N4S]2+ / 249.0964 / 249.0963 / 0.41
243 / [C24H30O5N4S]2+ / 243.0964 / 243.0966 / 0.42
211.6 / [C23H25O3N3S]2+ / 211.5803 / 211.5808 / 0.08
197.6 / [C22H25O2N3S]2+ / 197.5828 / 197.5834 / -0.13
130 / [C5H8O3N]+ / 130.0496 / 130.0499 / -2.1

Table S3. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M3 (Rt = 8.13 min) at m/z 280.6 (see Figure S3A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1846 / 485.1853 / -1.5
431 / [ C21H27O4N4S ]+ / 431.1746 / 431.1748 / -0.35
422 / [ C23H24O3N3S ]+ / 422.1531 / 422.1533 / -0.45
328 / [ C18H22ON3S ]+ / 328.1478 / 328.1484 / -0.03
311 / [ C18H19ON2S ]+ / 311.1213 / 311.1218 / 0.127
287 / [C16H19ON2S]+ / 287.1214 / 287.1213 / 0.49
285 / [C16H17ON2S]+ / 285.1057 / 285.1062 / 0.31
249 / [C25H30O5N4S]2+ / 249.0963 / 249.0963 / 0.01
243 / [C24H30O5N4S]2+ / 243.0963 / 243.0966 / 0.01
211.6 / [C23H25O3N3S]2+ / 211.5802 / 211.5808 / -0.39
197.6 / [C22H25O2N3S]2+ / 197.5827 / 197.5834 / -0.64
130 / [C5H8O3N]+ / 130.0495 / 130.0499 / -2.8

Table S4. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M4 (Rt = 7.72 min) at m/z 280.6 (see Figure S4A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1852 / 485.1853 / -0.24
431 / [ C21H27O4N4S ]+ / 431.1750 / 431.1748 / 0.57
422 / [ C23H24O3N3S ]+ / 422.1536 / 422.1533 / 0.74
328 / [ C18H22ON3S ]+ / 328.1481 / 328.1484 / 0.88
311 / [ C18H19ON2S ]+ / 311.1217 / 311.1218 / 1.4
287 / [C16H19ON2S]+ / 287.1217 / 287.1213 / 1.5
285 / [C16H17ON2S]+ / 285.1060 / 285.1062 / 1.4
249 / [C25H30O5N4S]2+ / 249.0966 / 249.0963 / 1.2
243 / [C24H30O5N4S]2+ / 243.0966 / 243.0966 / 1.2
211.6 / [C23H25O3N3S]2+ / 211.5804 / 211.5808 / 0.55
197.6 / [C22H25O2N3S]2+ / 197.5828 / 197.5834 / -0.13
130 / [C5H8O3N]+ / 130.0496 / 130.0499 / -2.1

Table S5. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M7 (Rt = 5.60 min) at m/z 280.6 (see Figure S5A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1847 / 485.1853 / -1.3
431 / [ C21H27O4N4S ]+ / 431.1744 / 431.1748 / -0.82
422 / [ C23H24O3N3S ]+ / 422.1531 / 422.1533 / -0.45
328 / [ C18H22ON3S ]+ / 328.1478 / 328.1484 / -0.03
311 / [ C18H19ON2S ]+ / 311.1214 / 311.1218 / 0.45
287 / [C16H19ON2S]+ / 287.1213 / 287.1213 / 0.14
285 / [C16H17ON2S]+ / 285.1058 / 285.1062 / 0.66
249 / [C25H30O5N4S]2+ / 249.0963 / 249.0963 / 0.01
243 / [C24H30O5N4S]2+ / 243.0963 / 243.0966 / 0.01
211.6 / [C23H25O3N3S]2+ / 211.5802 / 211.5808 / -0.39
197.6 / [C22H25O2N3S]2+ / 197.5826 / 197.5834 / -1.1
130 / [C5H8O3N]+ / 130.0495 / 130.0499 / -2.8

Table S6. List of accurate masses and chemical formulae of the product ions obtained by CID of

the molecular ion of M8 (Rt = 5.27 min) at m/z 280.6 (see Figure S6A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1847 / 485.1853 / -1.3
431 / [ C21H27O4N4S ]+ / 431.1745 / 431.1748 / -0.59
422 / [ C23H24O3N3S ]+ / 422.1534 / 422.1533 / 0.26
328 / [ C18H22ON3S ]+ / 328.1479 / 328.1484 / 0.28
311 / [ C18H19ON2S ]+ / 311.1212 / 311.1218 / -0.19
287 / [C16H19ON2S]+ / 287.1214 / 287.1213 / 0.49
285 / [C16H17ON2S]+ / 285.1057 / 285.1062 / 0.31
249 / [C25H30O5N4S]2+ / 249.0963 / 249.0963 / 0.01
243 / [C24H30O5N4S]2+ / 243.0963 / 243.0966 / 0.01
211.6 / [C23H25O3N3S]2+ / 211.5801 / 211.5808 / -0.86
197.6 / [C22H25O2N3S]2+ / 197.5827 / 197.5834 / -0.64
130 / [C5H8O3N]+ / 130.0495 / 130.0499 / -2.8

Table S7. List of accurate mass measurements and chemical formulae of the product ions obtained by CID of

the molecular ion of M9 (Rt = 4.60 min) at m/z 280.6 (see Figure S7A and 2)

Fragment ion

(m/z) / Chemical formula /

Measured mass

(Da) /

Calculated mass

(Da) /

Mass error

(ppm)
485 / [C24H29O5N4S]+ / 485.1847 / 485.1853 / -1.3
431 / [ C21H27O4N4S ]+ / 431.1744 / 431.1748 / -0.82
422 / [ C23H24O3N3S ]+ / 422.1529 / 422.1533 / -0.92
328 / [ C18H22ON3S ]+ / 328.1478 / 328.1484 / -0.03
311 / [ C18H19ON2S ]+ / 311.1214 / 311.1218 / 0.45
287 / [C16H19ON2S]+ / 287.1213 / 287.1213 / 0.14
285 / [C16H17ON2S]+ / 285.1057 / 285.1062 / 0.31
249 / [C25H30O5N4S]2+ / 249.0962 / 249.0963 / -0.39
243 / [C24H30O5N4S]2+ / 243.0962 / 243.0966 / -0.40
211.6 / [C23H25O3N3S]2+ / 211.5801 / 211.5808 / -0.86
197.6 / [C22H25O2N3S]2+ / 197.5826 / 197.5834 / -1.1
130 / [C5H8O3N]+ / 130.0495 / 130.0499 / -2.1

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