Supplementary Information - Semantic Scholar · S6 Figure S6: HR-MS spectra of compound 6b HR-MS...

S1

Supplementary Information

Methylthiodeoxynivalenol (MTD): Insight into chemistry, structure and

toxicity of thia-Michael adducts of trichothecenes

Philipp Fruhmann,a# Theresa Weigl-Pollack,a# Hannes Mikula,*a

Gerlinde Wiesenberger,b Gerhard Adam,b Elisabeth Varga,c Franz Berthiller,c Rudolf Krska,c

Christian Hametner,a Johannes Fröhlicha

a Institute of Applied Synthetic Chemistry, Vienna University of Technology (VUT), Getreidemarkt 9/163,

1060 Vienna, Austria.

E-mail: [email protected]

b Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences,

Vienna (BOKU), Konrad Lorenz Str. 24, 3430 Tulln, Austria.

c Christian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry,

Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Vienna

(BOKU), Konrad Lorenz Str. 20, 3430 Tulln, Austria.

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2014

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Table of Contents

1. NMR and HR-MS/MS spectra of 6b, 8, 9 and 11 S3

Figure S1: 1H NMR spectrum of compound 6b S3

Figure S2: 13C NMR spectrum of compound 6b S3

Figure S3: COSY spectrum of compound 6b S4

Figure S4: HSQC spectrum of compound 6b S4

Figure S5: HMBC spectrum of compound 6b S5

Figure S6: HRMS spectra of compound 6b S6

Figure S7: HRMS spectra of compound [6a] S7

Figure S8: 1H NMR spectrum of compound 8 S8

Figure S9: 13C NMR (APT) spectrum of compound 8 S8





2. Quantum Chemical Calculations S11

S3

1. NMR spectra of compounds 6b, 8, 9 and 11

Figure S1: 1H NMR spectrum of compound 6b

Figure S2: 13C NMR spectrum of compound 6b

11 10 9 8 7 6 5 4 3 2 1 ppm

NAME WPT051#FEXPNO 31PROCNO 1

Date_ 20120807Time 18.12

INSTRUM spectPROBHD 5 mm MultinuclPULPROG zg30

TD 32768SOLVENT MeOD

NS 32DS 0SWH 4789.272 Hz

FIDRES 0.146157 HzAQ 3.4210291 sec

RG 64DW 104.400 usecDE 6.00 usec

TE 296.2 KD1 1.00000000 sec

TD0 1

======== CHANNEL f1 ========

NUC1 1HP1 13.40 usec

PL1 6.00 dBSFO1 400.1322007 MHzSI 16384

SF 400.1300078 MHzWDW EM

SSB 0LB 0.30 HzGB 0

PC 1.00

220 200 180 160 140 120 100 80 60 40 20 0 ppm


Date_ 20120807Time 18.18INSTRUM spect

PROBHD 5 mm MultinuclPULPROG zgpg30

TD 65536SOLVENT MeODNS 24576

DS 4SWH 25125.629 Hz

FIDRES 0.383387 Hz

AQ 1.3042164 secRG 32768

DW 19.900 usecDE 6.00 usec

TE 296.2 KD1 1.00000000 secd11 0.03000000 sec

DELTA 0.89999998 secTD0 1

======== CHANNEL f1 ========NUC1 13C

P1 14.40 usecPL1 0.00 dB

SFO1 100.6238364 MHz

======== CHANNEL f2 ========

CPDPRG2 waltz16NUC2 1H

PCPD2 100.00 usecPL2 6.00 dBPL12 24.00 dB

PL13 26.00 dBSFO2 400.1316005 MHz

SI 32768SF 100.6126277 MHzWDW EM

SSB 0LB 1.00 Hz

GB 0

PC 1.40

1.52.02.53.03.54.04.5 ppm

S4

Figure S3: COSY spectrum of compound 6b

Figure S4: HSQC spectrum of compound 6b

S5

Figure S5: HMBC spectrum of compound 6b

S6

Figure S6: HR-MS spectra of compound 6b

HR-MS (-ESI) spectrum of 6b at 7.83 min

379.0994

341.1247

265.1084 295.1190

313.1121277.1086 406.1178

260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 4500

0.4

0.8

1.2

1.6

2.0

250

[MTD+HCOO]-

[MTD+Cl]-

389.1279

Counts (x106) vs. Mass-to-Charge (m/z)

HR-MS/MS (-ESI) product ion spectrum of 6b at 7.85 min, collision energy 10 eV

295.1192

341.1247

265.1084

247.0973 313.1117 389.1267138.0317

120 140 160 180 200 220 240 260 280 300 320 340 360 380


0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

100

[DON-H]-

[DON+HCOO]-

[MTD+HCOO]-

[DON-CH2O-H]-

[MTD-CH2O-H]-[DON-CH2O

-H2O-H]-

S7

Figure S7: HR-MS spectra of compound [6a]

HR-MS (-ESI) spectrum of [6a] at 8.66 min

389.1283

313.1121265.1087

343.1224


260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 4500

0.5

1.0

1.5

2.0

2.5

3.0[MTD+HCOO]-

[MTD+Cl]-

379.0992

250

HR-MS/MS (-ESI) product ion spectrum of [6a] at 8.62 min, collision energy 10 eV

313.1121

223.0805 389.1280295.1190

343.1223

265.1084

247.0979138.0323

120 140 160 180 200 220 240 260 280 300 320 340 360 380


0

0.4

0.8

1.2

1.6

2.0

100

[DON-H]-

[MTD-H]-

[MTD+HCOO]-

[DON-CH2O-H]-

[MTD-CH2O-H]-

[C12H16O2S-H]-

341.1242

[DON+HCOO]-

S8

Figure S8: 1H NMR spectrum of compound 8

Figure S9: 13C NMR (APT) spectrum of compound 8

11 10 9 8 7 6 5 4 3 2 1 ppm

NAME WPT057_1#FEXPNO 1PROCNO 1

Date_ 20120711Time 9.33


TD 32768SOLVENT CDCl3

NS 16DS 0SWH 4789.272 Hz

FIDRES 0.146157 HzAQ 3.4210291 sec


TE 296.2 KD1 1.00000000 sec

TD0 1

======== CHANNEL f1 ========

NUC1 1H

P1 13.40 usec

PL1 6.00 dBSFO1 400.1322007 MHzSI 16384

SF 400.1300092 MHzWDW no

SSB 0LB 0.00 HzGB 0

PC 1.00

260 240 220 200 180 160 140 120 100 80 60 40 20 0 ppm

NAME WPT007EXPNO 63PROCNO 1

Date_ 20120508Time 1.57

INSTRUM spectPROBHD 5 mm Dual 13C/PULPROG jmod


NS 2500DS 4SWH 15060.241 Hz

FIDRES 0.229801 HzAQ 2.1758451 sec

RG 16384DW 33.200 usec

DE 6.00 usec

TE 300.0 KCNST2 145.0000000

CNST11 1.0000000D1 3.00000000 secD20 0.00689655 sec

TD0 1

======== CHANNEL f1 ========NUC1 13CP1 9.00 usec

P2 18.00 usecPL1 0.00 dB

SFO1 50.3287677 MHz

======== CHANNEL f2 ========

CPDPRG2 waltz16

NUC2 1H

PCPD2 96.00 usecPL2 0.00 dBPL12 16.72 dB

SFO2 200.1308005 MHzSI 32768

SF 50.3227237 MHzWDW EMSSB 0

LB 1.00 HzGB 0

PC 1.40

1.11.21.31.41.51.61.71.81.92.02.12.22.32.42.52.6 ppm

S9



11 10 9 8 7 6 5 4 3 2 1 ppm


Date_ 20120709Time 11.47



NS 16

DS 0SWH 4789.272 Hz

FIDRES 0.146157 HzAQ 3.4210291 sec


TE 296.2 KD1 1.00000000 sec

TD0 1

======== CHANNEL f1 ========


PL1 6.00 dBSFO1 400.1322007 MHzSI 16384


SSB 0LB 0.00 HzGB 0

PC 1.00

260 240 220 200 180 160 140 120 100 80 60 40 20 0 ppm

NAME WPT007EXPNO 73PROCNO 1

Date_ 20120508Time 7.53INSTRUM spect

PROBHD 5 mm Dual 13C/PULPROG jmod

TD 65536SOLVENT CDCl3NS 4000

DS 4SWH 15060.241 Hz

FIDRES 0.229801 HzAQ 2.1758451 secRG 23170.5

DW 33.200 usec

DE 6.00 usec

TE 300.0 KCNST2 145.0000000CNST11 1.0000000

D1 3.00000000 secD20 0.00689655 sec

TD0 1

======== CHANNEL f1 ========

NUC1 13CP1 9.00 usec

P2 18.00 usecPL1 0.00 dBSFO1 50.3287677 MHz

======== CHANNEL f2 ========

CPDPRG2 waltz16NUC2 1H

PCPD2 96.00 usec

PL2 0.00 dBPL12 16.72 dB

SFO2 200.1308005 MHzSI 32768SF 50.3227225 MHz

WDW EMSSB 0

LB 1.00 HzGB 0PC 1.40

1.71.81.92.02.12.22.32.42.52.62.72.82.93.03.13.23.33.4 ppm

S10



11 10 9 8 7 6 5 4 3 2 1 ppm


Date_ 20120712Time 10.36



NS 16DS 0SWH 4789.272 Hz

FIDRES 0.146157 HzAQ 3.4210291 sec


TE 296.2 KD1 1.00000000 sec

TD0 1

======== CHANNEL f1 ========


PL1 6.00 dBSFO1 400.1322007 MHzSI 16384


SSB 0LB 0.00 Hz

GB 0

PC 1.00

220 200 180 160 140 120 100 80 60 40 20 0 ppm


Date_ 20120711

Time 21.40

INSTRUM spectPROBHD 5 mm MultinuclPULPROG jmod


NS 4096DS 4SWH 25125.629 Hz

FIDRES 0.383387 HzAQ 1.3042164 sec


TE 296.2 KCNST2 145.0000000

CNST11 1.0000000D1 3.00000000 secd20 0.00689655 sec

DELTA 0.00001833 sec

TD0 1

======== CHANNEL f1 ========NUC1 13C

P1 14.40 usecp2 28.80 usec

PL1 0.00 dBSFO1 100.6238364 MHz

======== CHANNEL f2 ========CPDPRG2 waltz16

NUC2 1HPCPD2 100.00 usec

PL2 6.00 dBPL12 24.00 dBSFO2 400.1316005 MHz

SI 32768SF 100.6127613 MHz

WDW EMSSB 0

LB 1.00 Hz

GB 0PC 1.40

1.11.21.31.41.51.61.71.81.92.02.12.22.32.42.52.62.7 ppm

S11

2. Quantum Chemical Calculations

Computational methods

Geometry optimisations to energy minima as well as subsequent frequency analyses of all considered

starting geometries were achieved applying the PM6 semiempirical method1 followed by DFT calculations2

at the level 6-311++G(d,p),3 using the B3LYP hybrid functional,4 as implemented in the Gaussian 09

(Revision A.1) program package.5 For calculations in methanol and water the polarisable continuum model

(PCM)6 using the integral equation formalism variant (IEFPCM) was applied. Data analysis was done using

GaussView 5 (Gaussian, Inc.).

Modelling of hemiketalisation

Table S1: Calculated energies for all compounds in gas phase and water.

Reaction Compound Eketo(a) [Hartree] Ehemiacetal(b) [Hartree]

gas phase water gas phase water

i I -423.3149 -423.3270 -423.3034 -423.3120

ii II -424.5429 -424.5525 -424.5385 -424.5464

iii III -769.2026 -769.2139 -769.1934 -769.2029

iv IV -1207.9545 -1207.9667 -1207.9547 -1207.9648

v 1 -1035.1823 -1035.1993 -1035.1748 -1035.1903

vi 6 -1473.9344 -1473.9506 -1473.9357 -1473.9509

1 Stewart, J. P. J. Mol. Model. 2007, 13, 1173-1213. 2 (a) Kohn, W.; Sham, L. J. Phys. Rev. 1965, 140, A1133-A1138. (b) Parr, R. G.; Yang, W. Density Functional

Theory of Atoms and Molecules. Oxford University Press: London, 1989. 3 Krishnan, R.; Binkley, J. S.; Seeger, R.; Pople, J. A. J. Chem. Phys. 1980, 72, 650-654. 4 Becke, A. D. J. Chem. Phys. 1993, 98, 5648-5652. 5 Frisch, M. J.; Trucks, G. W.; Schlegel, H. B.; Scuseria, G. E.; Robb, M. A.; Cheeseman, J. R.; Scalmani, G.;

Barone, V.; Mennucci, B.; Petersson, G. A.; Nakatsuji, H.; Caricato, M.; Li, X.; Hratchian, H. P.; Izmaylov, A. F.;

Bloino, J.; Zheng, G.; Sonnenberg, J. L.; Hada, M.; Ehara, M.; Toyota, K.; Fukuda, R.; Hasegawa, J.; Ishida, M.;

Nakajima, T.; Honda, Y.; Kitao, O.; Nakai, H.; Vreven, T.; Montgomery, J., J. A.; ; Peralta, J. E.; Ogliaro, F.;

Bearpark, M.; Heyd, J. J.; Brothers, E.; Kudin, K. N.; Staroverov, V. N.; Kobayashi, R.; Normand, J.;

Raghavachari, K.; Rendell, A.; Burant, J. C.; Iyengar, S. S.; Tomasi, J.; Cossi, M.; Rega, N.; Millam, J. M.; Klene,

M.; Knox, J. E.; Cross, J. B.; Bakken, V.; Adamo, C.; Jaramillo, J.; Gomperts, R.; Stratmann, R. E.; Yazyev, O.;

Austin, A. J.; Cammi, R.; Pomelli, C.; Ochterski, J. W.; Martin, R. L.; Morokuma, K.; Zakrzewski, V. G.; Voth, G.

A.; Salvador, P.; Dannenberg, J. J.; Dapprich, S.; Daniels, A. D.; Farkas, Ö.; Foresman, J. B.; Ortiz, J. V.;

Cioslowski, J.; Fox, D. J., Gaussian 09, Revision A.1. Gaussian Inc: Wallingford CT, 2009. 6 Tomasi, J.; Mennucci, B.; Cammi, R. Chem. Rev. 2005, 105, 2999-3094.

S12

Figure S14: Optimised geometries for hemiketalisation of model compounds I-IV, DON (1)

and MTD (6)

S13

Conformation of MTD (6b) in solution

On the basis of the observed NOESY correlations several starting geometries of MTD were optimised in

gas phase applying PM6 and DFT calculations. Lowest total energy was obtained for conformation 6b,

which was subsequently optimised in solution (MeOH, see Figure 2 in the manuscript).

Supplementary Information - Semantic Scholar · S6 Figure S6: HR-MS spectra of compound 6b HR-MS...

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