Supplementary Material

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[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.
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  

   

 

 

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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.  

     

 

 

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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.  

         

 

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