Supplementary Material Monitoring protein glycation by electrospray ionization (ESI) quadrupole time-‐of-‐flight (Q-‐TOF) mass spectrometer H. Gül Akıllıoğlu1, Ömür Çelikbıçak2, Bekir Salih2, Vural Gökmen1* 1 Food Quality and Safety (FoQuS) Research Group, Department of Food
Engineering, Hacettepe University, 06800 Beytepe Campus, Ankara, Turkey 2 Department of Chemistry, Hacettepe University, 06800 Beytepe Campus,
Ankara, Turkey Running Title: Monitoring protein glycation by ESI-‐Q-‐TOF MS * Corresponding Author Tel: +90-‐312-‐2977108; fax: +90-‐312-‐2992123; e-‐mail:
[email protected]
Table S1. Observed m/z values and peak areas of the signals of glycated cytochrome C (heated at 70 ᵒC for 3h in dry state). Observed ion for Cytochrome C Type
Observed m/z
Peak area
[M+18H]18+
a0
Not detected
N/A
[M+Glucose-‐H2O+18H]18+
a1
Not detected
N/A
[M+2Glucose-‐2H2O +18H]18+
a2
705.6850
402
[M+3Glucose-‐3H2O +18H]18+
a3
714.6468
695
[M+4Glucose-‐4H2O +18H]18+
a4
723.6548
988
[M+5Glucose-‐5H2O +18H]18+
a5
732.6696
1427
[M+6Glucose-‐6H2O +18H]18+
a6
741.6751
2034
[M+7Glucose-‐7H2O +18H]18+
a7
750.6833
3263
[M+8Glucose-‐8H2O +18H]18+
a8
759.6865
3237
[M+9Glucose-‐9H2O +18H]18+
a9
768.6991
3230
[M+10Glucose-‐10H2O +18H]18+
a10
777.7054
2031
[M+11Glucose-‐11H2O +18H]18+
a11
786.7236
1564
[M+12Glucose-‐12H2O +18H]18+
a12
795.7097
1348
[M+17H]17+
b0
Not detected
N/A
[M+Glucose-‐H2O +17H]17+
b1
Not detected
N/A
[M+2Glucose-‐2H2O +17H]17+
b2
Not detected
N/A
[M+3Glucose-‐3H2O +17H]17+
b3
756.6132
1619
[M+4Glucose-‐4H2O +17H]17+
b4
766.1646
2087
[M+5Glucose-‐5H2O +17H]17+
b5
775.7077
1903
[M+6Glucose-‐6H2O +17H]17+
b6
785.2461
2662
[M+7Glucose-‐7H2O +17H]17+
b7
794.7763
4250
[M+8Glucose-‐8H2O +17H]17+
b8
804.3179
4590
[M+9Glucose-‐9H2O +17H]17+
b9
813.8916
4929
[M+10Glucose-‐10H2O +17H]17+
b10
823.3848
3355
[M+11Glucose-‐11H2O +17H]17+
b11
832.9821
2744
[M+12Glucose-‐12H2O +17H]17+
b12
842.4652
1166
[M+16H]16+
c0
Not detected
N/A
[M+Glucose-‐H2O +16H]16+
c1
Not detected
N/A
[M+2Glucose-‐2H2O +16H]16+
c2
Not detected
N/A
[M+3Glucose-‐3H2O +16H]16+
c3
803.8809
113
[M+4Glucose-‐4H2O +16H]16+
c4
814.0016
425
[M+5Glucose-‐5H2O +16H]16+
c5
824.1309
1896
1
[M+6Glucose-‐6H2O +16H]16+
c6
834.2555
2956
[M+7Glucose-‐7H2O +16H]16+
c7
844.3874
3289
[M+8Glucose-‐8H2O +16H]16+
c8
854.5247
3221
[M+9Glucose-‐9H2O +16H]16+
c9
864.6528
3745
[M+10Glucose-‐10H2O +16H]16+
c10
874.7924
2228
[M+11Glucose-‐11H2O +16H]16+
c11
884.9279
2396
[M+12Glucose-‐12H2O +16H]16+
c12
895.0728
1137
[M+15H]15+
d0
Not detected
N/A
[M+Glucose-‐H2O +15H]15+
d1
Not detected
N/A
M+2Glucose-‐2H2O +15H]15+
d2
846.6098
125
[M+3Glucose-‐3H2O +15H]15+
d3
857.2817
628
[M+4Glucose-‐4H2O +15H]15+
d4
868.2098
1442
[M+5Glucose-‐5H2O +15H]15+
d5
878.9905
1881
[M+6Glucose-‐6H2O +15H]15+
d6
889.8056
2309
[M+7Glucose-‐7H2O +15H]15+
d7
900.6155
2654
[M+8Glucose-‐8H2O +15H]15+
d8
911.4211
2710
[M+9Glucose-‐9H2O +15H]15+
d9
922.2073
2376
[M+10Glucose-‐10H2O +15H]15+
d10
933.0455
2694
[M+11Glucose-‐11H2O +15H]15+
d11
943.8809
1523
[M+12Glucose-‐12H2O +15H]15+
d12
954.6060
1055
[M+14H]14+
e0
Not detected
N/A
[M+Glucose-‐ H2O +14H]14+
e1
Not detected
N/A
[M+2Glucose-‐2H2O +14H]14+
e2
Not detected
N/A
[M+3Glucose-‐3H2O +14H]14+
e3
918.4719
91
[M+4Glucose-‐4H2O +14H]14+
e4
930.1367
990
[M+5Glucose-‐5H2O +14H]14+
e5
941.7085
1288
[M+6Glucose-‐6H2O +14H]14+
e6
953.2914
1716
[M+7Glucose-‐7H2O +14H]14+
e7
964.8720
1791
[M+8Glucose-‐8H2O +14H]14+
e8
976.4494
2618
[M+9Glucose-‐9H2O +14H]14+
e9
988.0250
1537
[M+10Glucose-‐10H2O +14H]14+
e10
999.6271
1226
[M+11Glucose-‐11H2O +14H]14+
e11
1011.2081
887
[M+12Glucose-‐12H2O +14H]14+
e12
1022.7774
647
[M+13H]13+
f0
Not detected
N/A
[M+Glucose-‐H2O +13H]13+
f1
Not detected
N/A
[M+2Glucose-‐2H2O +13H]13+
f2
Not detected
N/A
2
[M+3Glucose-‐3H2O +13H]13+
f3
989.1047
518
[M+4Glucose-‐4H2O +13H]13+
f4
1001.6430
619
[M+5Glucose-‐5H2O +13H]13+
f5
1014.0579
745
[M+6Glucose-‐6H2O +13H]13+
f6
1026.5432
1056
[M+7Glucose-‐7H2O +13H]13+
f7
1039.0084
1109
[M+8Glucose-‐8H2O +13H]13+
f8
1051.4787
1672
[M+9Glucose-‐9H2O +13H]13+
f9
1063.9628
1036
[M+10Glucose-‐10H2O +13H]13+
f10
1076.4295
771
[M+11Glucose-‐11H2O +13H]13+
f11
1088.8948
496
[M+12Glucose-‐12H2O +13H]13+
f12
1101.3482
501
3
Table S2. Observed m/z values and peak areas for most abundant signals of β-‐ casein (A1 and A2 variants) (heated at 70 ᵒC for 1h in dry state). Observed ion for β-‐casein
Type
Observed m/z
Peak area
[A1+20H]20+
a0
1200.1662
760
[A2+20H]20+
a0*
1202.0711
823
[A1+Glucose-‐H2O+20H]20+
a1
1208.3225
1166
[A2+Glucose-‐H2O+20H]20+
a1*
1210.2399
1109
[A1+2Glucose-‐2H2O +20H]20+
a2
1216.3302
2577
[A2+2Glucose-‐2H2O +20H]20+
a2*
1218.3439
2325
[A1+3Glucose-‐3H2O +20H]20+
a3
1224.4588
2789
[A2+3Glucose-‐3H2O +20H]20+
a3*
1226.4735
2513
[A1+4Glucose-‐4H2O +20H]20+
a4
1232.5756
3327
[A2+4Glucose-‐4H2O +20H]20+
a4*
1234.5698
3012
[A1+5Glucose-‐5H2O +20H]20+
a5
1240.6953
2865
[A2+5Glucose-‐5H2O +20H]20+
a5*
1242.6781
2672
[A1+6Glucose-‐6H2O +20H]20+
a6
1248.7474
2620
[A2+6Glucose-‐6H2O +20H]20+
a6*
1250.7752
2541
[A1+7Glucose-‐7H2O +20H]20+
a7
1256.9319
1277
[A2+7Glucose-‐7H2O +20H]20+
a7*
1258.9102
1377
[A1+8Glucose-‐8H2O +20H]20+
a8
1265.0262
878
[A2+8Glucose-‐8H2O +20H]20+
a8*
1267.0173
563
[A1+19H]19+
b0
1263.2512
1357
[A2+19H]19+
b0*
1264.3527
1054
[A1+Glucose-‐H2O +19H]19+
b1
1270.7813
1172
[A2+Glucose-‐H2O +19H]19+
b1*
1272.8871
1066
[A1+2Glucose-‐2H2O +19H]19+
b2
1279.3188
1288
[A2+2Glucose-‐2H2O +19H]19+
b2*
1281.4313
1480
[A1+3Glucose-‐3H2O +19H]19+
b3
1287.9113
2061
[A2+3Glucose-‐3H2O +19H]19+
b3*
1289.9492
1893
[A1+4Glucose-‐4H2O +19H]19+
b4
1296.4001
3774
[A2+4Glucose-‐4H2O +19H]19+
b4*
1298.4881
3538
[A1+5Glucose-‐5H2O +19H]19+
b5
1304.9315
2126
[A2+5Glucose-‐5H2O +19H]19+
b5*
1307.0378
2107
[A1+6Glucose-‐6H2O +19H]19+
b6
1313.4534
1683
[A2+6Glucose-‐6H2O +19H]19+
b6*
1315.5372
1510
4
[A1+7Glucose-‐7H2O +19H]19+
b7
1321.9953
1187
[A2+7Glucose-‐7H2O +19H]19+
b7*
1324.0910
1402
[A1+8Glucose-‐8H2O +19H]19+
b8
1331.5286
634
[A2+8Glucose-‐8H2O +19H]19+
b8*
1333.6399
472
[A1+18H]18+
c0
1333.4090
1107
[A2+18H]18+
c0*
1335.6112
681
[A1+Glucose-‐H2O +18H]18+
c1
1342.4123
1164
[A2+Glucose-‐H2O +18H]18+
c1*
1344.6016
1079
[A1+2Glucose-‐2H2O +18H]18+
c2
1351.4120
2675
[A2+2Glucose-‐2H2O +18H]18+
c2*
1353.6343
2682
[A1+3Glucose-‐3H2O +18H]18+
c3
1360.4286
3383
[A2+3Glucose-‐3H2O +18H]18+
c3*
1362.6376
3416
[A1+4Glucose-‐4H2O +18H]18+
c4
1369.4366
3579
[A2+4Glucose-‐4H2O +18H]18+
c4*
1371.6475
3648
[A1+5Glucose-‐5H2O +18H]18+
c5
1378.4432
4312
[A2+5Glucose-‐5H2O +18H]18+
c5*
1380.6522
4109
[A1+6Glucose-‐6H2O +18H]18+
c6
1387.4601
3001
[A2+6Glucose-‐6H2O +18H]18+
c6*
1389.6674
3219
[A1+7Glucose-‐7H2O +18H]18+
c7
1396.4652
1756
[A2+7Glucose-‐7H2O +18H]18+
c7*
1398.6760
1867
[A1+8Glucose-‐8H2O +16H]18+
c8
1405.4717
983
[A2+8Glucose-‐8H2O +16H]18+
c8*
1407.6816
895
5
Figure S1. Deconvoluted ESI-‐QTOF mass spectra of cytochrome C and glucose mixture heated in solution at 70 ᵒC for 6h (A) Full deconvoluted mass spectrum (B) Expanded view of the region m/z 600–1400 (C) Expanded view of the region m/z 10500–14500, represents deconvoluted signals of glycated protein species.
6
Figure S2. Deconvoluted ESI-‐QTOF mass spectra of cytochrome C and glucose heated in dry state at 70 ᵒC for 6h (A) Full deconvoluted mass spectrum (B) Expanded view of the region m/z 720 –1300 (C) Expanded view of the region m/z 13000–15200, represents deconvoluted signals of glycated protein species.
7