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Supplementary Data
Elucidation of several neglected reactions in the GC-MS identification of sialic acids as
heptafluorobutyrates calls for an urgent reassessment of previous claims Paola Rota*, Luigi Anastasia, Pietro Allevi Department of Biomedical, Surgical and Dental Sciences, University of Milan, via Saldini 50, I-20133 Milan, Italy; corresponding
author: Paola Rota; Telephone: +39 02 50316047; Fax: +39 02 50316040; E-mail:[email protected]
Contents Page Number
1H and 13C NMR of Sias esters 1b and 1c S2-S3
1H and 13C NMR of 1,7 lactones 7b, 4b, 7c, 7d, 7e, 4c S4-S9
1H and 13C NMR of crude 1,7 lactones 8b, 8b S10-S11
1H and 13C NMR of crude 1,7 lactones 9a-c S12-S14
GC profile of HFB derivatives of Sias esters 1a-c S15
EI mass spectra of lactones 8a-c S16-S17
EI mass spectra of methyl esters of sialic acids 1a-c S18-S19
EI mass spectra of lactones 9a-c S20-S21
S-1
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2015
1.05
3.00
3.00
0.93
0.98
4.25
0.99
2.00
1.05
1.00
102030405060708090100110120130140150160170180190200f1 (ppm)
None 13C-NMR
methyl ester of 1b
S-2
S-3
S-4
20.7
22.3
30.3
50.2
64.8
65.9
68.4
70.1
77.7
90.4
168.9
169.0
170.3
S-5
compound 7c
1H-NMR
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)
None
22.4
25.5
27.4
37.0
52.4
67.3
67.8
71.4
73.6
75.6
80.7
94.9
111.2
129.4
129.7
129.8
136.3
153.5
167.5
172.9
S-6
S-7
S-8
3.59
3.67
3.04
1.41
2.16
1.33
2.15
1.05
1.07
0.99
0.92
1.00
S-9
3.35
2.13
1.34
1.28
0.97
2.17
1.13
1.00
0.72
20.1
32.5
48.7
61.1
70.1
70.4
71.4
74.4
76.0
93.2
160.9
173.0
S-10
0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)
None
compound 8c
1H-NMR
20.3
20.4
32.8
49.0
61.2
66.6
71.8
74.4
76.2
93.9
161.8
169.7
172.8
S-11
S-12
S-13
3.42
3.57
3.25
1.21
1.13
2.32
1.11
2.33
1.17
1.15
1.00
20.4
22.1
32.6
48.7
61.6
67.2
72.1
74.9
76.5
94.0
162.3
170.3
173.4
S-14
A)
B)
C)
Fig. 1S.Total ion chromatograms of products obtained by HFBAA derivatization (HFBAA, MeCN, 150°C, 5 min) of (A)
methyl ester of acid 1a, (B) methyl ester of acid 1b and (C) methyl ester of acid 1c.
S-15
0 1 2 3 4 5 6 7 8 9 10 11
Time (min)
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
7.80
8.84
10.818.02 10.187.216.712.70 5.943.33 5.574.71
0 1 2 3 4 5 6 7 8 9 10 11
Time (min)
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
9.00
10.7710.38
8.877.736.752.79 3.70 6.555.704.68
RT: 0.00 - 11.02
0 1 2 3 4 5 6 7 8 9 10 11
Time (min)
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
8.51
10.193.60
10.328.98 9.928.283.76 4.22 7.856.745.79
EI mass spectrum of lactone 8a obtained by derivatization (HFBAA, MeCN, 150 °C, 5 min) of the
corresponding acid 1a or of the lactone 4a
EI mass spectrum of lactone 8b obtained by derivatization (HFBAA, MeCN, 150 °C, 5 min) of the
corresponding acid 1b or of the lactone 4b
S-16
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
169
490276
292238 330197 758119 1017704492 534 774464374 968588
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
20
25
Re
lative
Ab
un
da
nce
169 490276
264
292
320238
197 758119 346 1017704492534347 560 774 802464 1017757 968451
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
169
490
264252
292 34781 119 491352 862543197 757451
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
Re
lative
Ab
un
da
nce
169 490264292 347238
81 119
491 531352 862543197
560363757451 863364 604 802648 834 906745
EI mass spectrum of lactone 8c obtained by derivatization (HFBAA, MeCN, 150 °C, 5 min) of the
corresponding acid 1c or of the lactone 4c.
S-17
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
337 490
295169
277 320
27681 338 491238 708119 543214 380 666603477406 902802752
100 200 300 400 500 600 700 800 900 1000
m/z
0
10
20
30
40
Re
lative
Ab
un
da
nce
337 490295169277
320
276
81 252 338491
82 238 352 708119 543214 380 666603 709550477406 902802
EI mass spectra obtained after derivatization (HFBAA, MeCN, 150 °C, 5 min) of the methyl ester of sialic acid
1a. A and B represent the mass spectra of peaks at 7.85 and 8.86 min of derivative of methyl ester of 1a
S-18
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
169
356119884 994175
388328 570424 626218 840253 784 885 1034670456 538
100 200 300 400 500 600 700 800 900 1000
m/z
0
10
20
30
Re
lative
Ab
un
da
nce
169356
119
116 884 994175
388191328 570424 626218 840
253 784 885638 1034670 856456 538
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
169
670
244200 356
144130 246 839
212 671328260 456 626413 820 840780594550
100 200 300 400 500 600 700 800 900 1000
m/z
0
10
20
30
Re
lative
Ab
un
da
nce
169 670244 356
144
130246
839
212 671328260 456 626413 820 840
300 808672424 625457 627 841550
A
B
B
A
EI mass spectra obtained after derivatization (HFBAA, MeCN, 150 °C, 5 min) of the methyl ester of sialic acid
1b. C represent the mass spectrum of peak at 9.00 min of derivative of methyl ester of 1b.
EI mass spectra obtained after derivatization (HFBAA, MeCN, 150 °C, 5 min) of the methyl ester of sialic acid
1c. D represent the mass spectrum of peak at 8.51 min of derivative of methyl ester of 1c.
S-19
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
150
169
131
207
192 281228 730356 899341 388 429 840456 686566 626503 780
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
20
25
30
Re
lative
Ab
un
da
nce
150 169131
192 281
228
730356
899341 388 429
840731456 900316 686566 626503 780
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
169
96 566294122 181 280 336 388 567 764608406 490 704
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
20
Re
lative
Ab
un
da
nce
16996 566
565
294122
181280131 336239 388 567
267 764608234 406 490 704448
C
D
D
C
EI mass spectra of lactone 9a obtained by derivatization with HFBAA at 23°C the acids 1a and the lactones 4a.
EI mass spectra of lactone 9b obtained by derivatization with HFBAA at 23°C the acids 1b and the lactones 4b.
S-20
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
96
169
320
124533
347
648252133 197 348 620306 534406 649520434
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
20
25
Re
lative
Ab
un
da
nce
96 169 320 533347
648252133
197 253 348 620534239 406281 649561 603520434
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
96
169124
133
379193 347252197 306 380 452 494
707
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
20
25
Re
lative
Ab
un
da
nce
96 169133379193 347
252194
306207
380 452 494253210408 493
495 707
EI mass spectra of lactone 9c obtained by derivatization with HFBAA at 23°C the acids 1c and the lactones 4c.
S-21
100 200 300 400 500 600 700 800 900 1000
m/z
0
20
40
60
80
100
Re
lative
Ab
un
da
nce
101
294102
142
143
336194134252 493
512351 554451396298 724682648 767
100 200 300 400 500 600 700 800 900 1000
m/z
0
5
10
15
Re
lative
Ab
un
da
nce
101 294142 169 336194
134
252 493295226296254
512351554451396298 724707553 665470 555 767
