si tx-com-03-2014-000019.r1 proofs · 4 5-(hydroxymethyl)furfural, hmf (1)1 to a round-bottom flask...

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Supplementary Information

Emerging Platform from Renewable Resources: Selection Guidelines for Human

Exposure of Furfural‐related Compounds

Raquel F. M. Frade*,a, Jaime A. S. Coelhoa, Svilen P. Simeonova, Carlos A. M. Afonso*,a

a Instituto de Investigação do Medicamento (iMed.ULisboa), Faculty of Pharmacy, University of Lisbon, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal

Procedures 3 5-(Hydroxymethyl)furfural, HMF (1) ................................................................................................................................... 4 Sodium (5-formylfuran-2-yl)methyl sulfate (2) .................................................................................................................... 4 5-(Chloromethyl)furan-2-carbaldehyde (3) .......................................................................................................................... 5 5-(Ethoxymethyl)furan-2-carbaldehyde (4) .......................................................................................................................... 5 5-((Benzyloxy)methyl)furan-2-carbaldehyde (5).................................................................................................................. 6 5-(((tert-Butyldimethylsilyl)oxy)methyl)furan-2-carbaldehyde (6) ...................................................................................... 6 (5-Formylfuran-2-yl)methyl acetate (7) ................................................................................................................................ 7 (5-Formylfuran-2-yl)methyl hexanoate (8) ........................................................................................................................... 7 (5-Formylfuran-2-yl)methyl benzoate (9) ............................................................................................................................. 8 Furan-2,5-dicarbaldehyde (10) ............................................................................................................................................. 8 Furan-2-carbaldehyde (11) ................................................................................................................................................... 9 Furan-2,5-diyldimethanol (12) .............................................................................................................................................. 9 (5-(Ethoxymethyl)furan-2-yl)methanol (13) ......................................................................................................................... 9 Sodium 5-(hydroxymethyl)furan-2-carboxylate (14) ......................................................................................................... 10 Furan-2-ylmethanol (15) ..................................................................................................................................................... 10 2,5-Bis(ethoxymethyl)furan (16) ........................................................................................................................................ 10 Furan-2,5-dicarboxylic acid, FDCA (17) ............................................................................................................................ 11 2,5-Dimethylfuran (18) ....................................................................................................................................................... 11 2-Methylfuran (19) ............................................................................................................................................................. 11 (5,5'-(Oxybis(methylene))bis(furan-5,2-diyl))dimethanol (20) .......................................................................................... 11 (5,5'-((1E,1'E)-hydrazine-1,2-diylidenebis(methanylylidene))bis(furan-5,2-diyl))dimethanol (21) ................................... 12 4-Oxopentanoic acid, levulinic acid (22) ............................................................................................................................ 12 Figures (HPLC, GC-MS and NMR) 13 Figure 1. HPLC chromatogram of 1. .................................................................................................................................. 13 Figure 2. HPLC chromatogram of 2. .................................................................................................................................. 13 Figure 3. HPLC chromatogram of 3. .................................................................................................................................. 14 Figure 4. HPLC chromatogram of 4. .................................................................................................................................. 14 Figure 5. HPLC chromatogram of 5. .................................................................................................................................. 15 Figure 6. HPLC chromatogram of 6. .................................................................................................................................. 15 Figure 7. HPLC chromatogram of 7. .................................................................................................................................. 16 Figure 8. HPLC chromatogram of 8. .................................................................................................................................. 16 Figure 9. HPLC chromatogram of 9. .................................................................................................................................. 17 Figure 10. HPLC chromatogram of 10. .............................................................................................................................. 17

Electronic Supplementary Material (ESI) for Toxicology Research.This journal is © The Royal Society of Chemistry 2014

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Figure 11. HPLC chromatogram of 11. .............................................................................................................................. 18 Figure 12. HPLC chromatogram of 13. .............................................................................................................................. 18 Figure 13. HPLC chromatogram of 16. .............................................................................................................................. 19 Figure 14. HPLC chromatogram of 17. .............................................................................................................................. 19 Figure 15. HPLC chromatogram of 18. .............................................................................................................................. 20 Figure 16. HPLC chromatogram of 19. .............................................................................................................................. 20 Figure 17. HPLC chromatogram of 20. .............................................................................................................................. 21 Figure 18. GC-MS chromatogram and MS of major signal of 1. ....................................................................................... 22 Figure 19. GC-MS chromatogram and MS of major signal of 3. ....................................................................................... 22 Figure 20. GC-MS chromatogram and MS of major signal of 6. ....................................................................................... 23 Figure 21. GC-MS chromatogram and MS of major signal of 7. ....................................................................................... 24 Figure 22. GC-MS chromatogram and MS of major signal of 11. ..................................................................................... 25 Figure 23. GC-MS chromatogram and MS of major signal of 12. ..................................................................................... 25 Figure 24. GC-MS chromatogram and MS of major signal of 15. ..................................................................................... 26 Figure 25. GC-MS chromatogram and MS of the major signals of 18. .............................................................................. 27 Figure 26. GC-MS chromatogram and MS of the major signals of 19. .............................................................................. 28 Figure 27. 1H NMR spectrum of 1. ..................................................................................................................................... 29 Figure 28. 13C NMR spectrum of 1. .................................................................................................................................... 30 Figure 29. 1H NMR spectrum of 2. ..................................................................................................................................... 31 Figure 30. 13C NMR spectrum of 2. .................................................................................................................................... 32 Figure 31. 1H NMR spectrum of 3. ..................................................................................................................................... 33 Figure 32. 1H NMR spectrum of 4. ..................................................................................................................................... 34 Figure 33. 13C NMR spectrum of 4. .................................................................................................................................... 35 Figure 34. 1H NMR spectrum of 5. ..................................................................................................................................... 36 Figure 35. 13C NMR spectrum of 5. .................................................................................................................................... 37 Figure 36. 1H NMR spectrum of 6. ..................................................................................................................................... 38 Figure 37. 13C NMR spectrum of 6. .................................................................................................................................... 39 Figure 38. 1H NMR spectrum of 7. ..................................................................................................................................... 40 Figure 39. 13C NMR spectrum of 7. .................................................................................................................................... 41 Figure 40. 1H NMR spectrum of 8. ..................................................................................................................................... 42 Figure 41. 13C NMR spectrum of 8. .................................................................................................................................... 43 Figure 42. 1H NMR spectrum of 9. ..................................................................................................................................... 44 Figure 43. 1H NMR spectrum of 10. ................................................................................................................................... 45 Figure 44. 13C NMR spectrum of 10. .................................................................................................................................. 46 Figure 45. 1H NMR spectrum of 12. ................................................................................................................................... 47 Figure 46. 13C NMR spectrum of 12. .................................................................................................................................. 48 Figure 47. 1H NMR spectrum of 13. ................................................................................................................................... 49 Figure 48. 1H NMR spectrum of 16. ................................................................................................................................... 50 Figure 49. 1H NMR spectrum of 17. ................................................................................................................................... 51 Figure 50. 1H NMR spectrum of 20. ................................................................................................................................... 52 Figure 51. 13C NMR spectrum of 20. .................................................................................................................................. 53 Figure 52. 1H NMR spectrum of 21. ................................................................................................................................... 54 Figure 53. 13C NMR spectrum of 21. .................................................................................................................................. 55 Toxicity curves 56 References 67

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Procedures General Remarks

All solvents were freshly distilled from commercial grade sources. Commercially available reagents were

used as received without further purification otherwise notice. Preparative thin-layer chromatography plate

was prepared with silica gel 60 GF254 MercK (ref 1.07730.1000), whereas flash chromatography was carried

out on silica gel 60M purchased from MN (Ref. 815381). Reaction mixtures were analyzed by TLC using

ALUGRAM SIL G/UV254 from MN (Ref. 818133, silica gel 60), and visualization by UV and

phosphomolybdic acid stain.

HPLC analysis was performed on Dionex P680 pump, Dionex UVD 340S diode array detector, detection

at 275 and 225 nm, manual injector with 20 μL loop, column HICHROM C18, 250x4.6mm, or Kromasil 100,

C18, 250x4.6mm. Mobile phase gradient from 1:99 to 50:50 for 40 min acetonitrile:water, and then 50:50 for

the time indicated in the chromatogram, flow 1 mL/min, The HPLC purity was determined by comparing the

integration area of the main signal with other observed minor peaks and are represented in the chromatograms

by relative area. Retention times were determined by using the mobile phase gradient from 1:99 to 90:10 in 50

min.

GC–MS analyses were performed on Gas Chromatograph Mass Spectrometer-QP2010S, Shimadzu by

using the column TRB-5MS-Teknokroma (30 m × 0.25 mm × 0.25 µm). GC program: column oven Tinitial=:

50.0 °C, Tfinal=: 250.0 °C, slope = 5ºC/min ; injection temperature: 250 °C; pressure: 77.9 kPa, total flow: 17.7

mL/min; column flow: 1.34 mL/min; linear velocity: 42.0 cm/sec; purge flow: 3.0 mL/min split ratio: 10.0,

high press. inj. pressure: 100.0 kPa, high press. inj. time: 1.00 min. MS program: start time: 3.00 min; end

time: 50.00 min; event time: 0.50 s; scan speed: 666; start: m/z = 40.00; end: m/z = 350.00.

NMR spectra were recorded at room temperature in a Bruker AMX 300 or Bruker AMX 400 using CDCl3,

D2O or DMSO-d6 as solvents and (CH3)4Si(1H) as internal standard.

4

5-(Hydroxymethyl)furfural, HMF (1)1

To a round-bottom flask D-fructose (6 g, 33.3 mmol) and DMSO (200 mL) were added followed by the

addition of Amberlyst-15-powder (0.4 g, 15% w/w). The reaction was allowed to stir for 120 minutes at 120

ºC under inert atmosphere. After reaction the DMSO was removed by distillation and the crude reaction

mixture was purified by column chromatography with silica gel (hexane/ethyl acetate 1:1) to yield the product

(2.9 g, 70% yield) as a pale yellow oil that crystalizes in the freezer (0ºC). 1H NMR (300 MHz, CDCl3) δ 2.59

(s, 1H), 4.71 (s, 2H), 6.51 (d, J = 3.5 Hz, 1H), 7.21 (d, J = 3.5 Hz, 1H), 9.58 (s, 1H); 13C NMR (100 MHz,

CDCl3) δ 57.7, 110.1, 123.1, 152.4, 160.8, 177.9.

As alternative, HMF was synthetized using the recently reported protocol from our group, using

tetraethylammonium bromide and Amberlyst-15.2

Sodium (5-formylfuran-2-yl)methyl sulfate (2)3

To a solution of N,N'-Dicyclohexylcarbodiimide (1.0 g, 5 mmol) in DMF (3 mL) at 0 ºC, HMF (128 mg, 1

mmol) dissolved in DMF (0.5 mL) was added with vigorous stirring. A cold solution of sulfuric acid (2 mmol)

in DMF (1 mL) was added. The mixture was allowed to stir at 0 ºC for 1h. The mixture was then centrifuged

and the solution neutralized with methanolic NaOH 1M. The resulting solution was once again centrifuged.

The precipitate was washed with DMF and centrifuged. The collected organic phases were concentrated under

vacuum and the crude dissolved in ethanol and centrifuged. To the solution 10 volumes of Et2O was slowly

added and the precipitate centrifuged. The precipitate was then washed with Et2O to give the product as a pale

orange solid (55 mg, 24% yield); mp 128 ºC.

5

5-(Chloromethyl)furan-2-carbaldehyde (3)4

OH

O

Cl

To a flame dried round bottom flask, HMF (1 g, 7.9 mmol) and CHCl3 were added. Trimethylsilyl chloride

(6 mL, 47.3 mmol, 6 equiv.) was added dropwise at 45 ºC and the resulting mixture stirred for 24 h at 45ºC

under inert atmosphere. The mixture was then quenched with saturated aqueous solution of NaHCO3 and

extracted with DCM. The collected organic layers were dried over Na2SO4 and the solvent removed. The

crude product was purified by dissolution in hot hexane and filtration through a pad of celite and activated

carbon. The hexane was removed under vacuum to give the desired product (0.7 g, 61% yield). 1H NMR (300

MHz, CDCl3) δ 4.59 (s, 2H), 6.57 (d, J = 3.57 Hz, 1H), 7.19 (d, J = 3.57 Hz, 1H), 9.60 (s, 1H).

5-(Ethoxymethyl)furan-2-carbaldehyde (4)5

HMF (1.0 g, 8.0 mmol) was dissolved in absolute EtOH (15 mL) then 3 drops of concentrated H2SO4 were

added and mixture was stirred for 5 h at reflux temperature. The reaction mixture was neutralized with

aqueous saturated solution of NaHCO3 and extracted with EtOAc. The organic phase was dried with MgSO4

and evaporated. The product was purified by silica flash chromatography using EtOAc/Hexane to give 703

mg (57% yield) of the desired product. 1H NMR (300 MHz, CDCl3) δ 1.20 (t, J = 7.0 Hz, 3H), 3.55 (q, J = 7

Hz, 2H), 4.49 (s, 2H), 6.49 (d, J = 3.52 Hz, 1H), 7.18 (d, J = 3.52 Hz, 1H), 9.56 (s, 1H); 13C NMR (100 MHz,

CDCl3) δ 15.1, 64.8, 66.7, 111.1, 152.6, 158.8, 177.9.

6

5-((Benzyloxy)methyl)furan-2-carbaldehyde (5)6

Benzyl bromide (3.8 g, 22.2 mmol) and silver oxide (2.6 g, 11.1 mmol) were successively added to a

stirred solution of HMF (1.4 g, 11.1 mmol) dissolved in DMF (15 mL). The mixture was stirred for 50 h at

RT. The solution was evaporated under reduced pressure and the residue was chromatographed (silica;

hexane-EtOAc, 1:1) to give the desired product (1.2 g, 50 % yield). 1H NMR (300 MHz, CDCl3) δ 0.87 (t, J =

6.9 Hz, 3H), 1.27-1.31 (m, 4H), 1.63 (quint, J = 7.4 Hz, 2H), 2.35 (t, J = 7.6, 2H), 5.12 (s, 2H), 6.58 (d, J =

3.55 Hz, 1H), 7.21 (d, J = 3.55 Hz, 1H), 9.63 (s, 1H); 13C NMR (100 MHz, CDCl3) δ 64.2, 73.0, 111.4, 122.0,

128.0, 128.1, 128.6, 137.3, 152.7, 158.5, 177.8.

5-(((tert-Butyldimethylsilyl)oxy)methyl)furan-2-carbaldehyde (6)7

To a solution of HMF (1.6 g, 12.7 mmol) in DMF (3.2 mL) were added imidazole (2.2 g, 31.7 mmol, 2.5

equiv.) and tert-butyldimethylsilyl chloride (2.3 g, 15.2 mmol, 1.2 equiv.). The solution was stirred for 22 h.

The silylated compound was extracted with hexane (5 x 20 mL). The collected organic layers were washed

with brine, dried over Na2SO4, and the solvent removed to give the crude product that was distillated

(100ºC/0.8 mbar) giving the product (1.9 g, 63 % yield) as a light yellow liquid that solidifies in the freezer

(0ºC). 1H NMR (300 MHz, CDCl3) δ 0.08 (s, 6H), 0.89 (s, 9H), 4.71 (s, 2H), 6.45 (d, J = 3.53 Hz, 1H), 7.19

(d, J = 3.53 Hz, 1H), 9.56 (s, 1H); 13C NMR (100 MHz, CDCl3) δ -5.3, 18.4, 25.8, 58.7, 109.5, 122.6, 152.3,

161.5, 175.6.

7

(5-Formylfuran-2-yl)methyl acetate (7)6

To a flame dried round-bottom flask, HMF (3.5 g, 27.8 mmol), MeCN (50 mL) and acetic anhydride (4.3

mL, 45.6 mmol) were added followed by catalytic amount of pyridine (0.5 mL, 6.2 mmol). The reaction

mixture was allowed to stir at room temperature for 22h under argon atmosphere. The solvent was removed

and the product purified by column chromatography with silica gel (hexane/ethyl acetate 8:2) to give the

product (4.3 g, 93% yield) as a pale yellow oil that crystallized in the freezer (0ºC). 1H NMR (300 MHz,

CDCl3) δ 2.11 (s, 3H), 5.12 (s, 2H), 6.58 (d, J = 3.55 Hz, 1H), 7.21 (d, J = 3.55 Hz, 1H), 9.64 (s, 1H); 13C

NMR (100 MHz, CDCl3) δ 20.8, 57.9, 112.7, 121.8, 153.0, 155.6, 170.5, 178.0.

(5-Formylfuran-2-yl)methyl hexanoate (8)8

To a flame dried round-bottom flask, HMF (0.250 g, 1.98 mmol), MeCN (10 mL) and hexanoic anhydride

(0.51 mL, 2.2 mmol, 1.1 equiv.) were added followed by catalytic amount of pyridine (32 µL, 0.4 mmol, 0.2

equiv.). The reaction mixture was allowed to stir at room temperature for 66 h under argon atmosphere. The

mixture was quenched with cold water, acidified with HCl 1M and extracted with Et2O. The combined

organic layers were dried over MgSO4, filtered and the solvent removed to give the crude product that was

purified by column chromatography with silica gel (hexane/ethyl acetate 8:2) to give the product (0.315 g,

71% yield) as a pale yellow oil. 1H NMR (300 MHz, CDCl3) δ 0.87 (t, J = 6.9 Hz, 3H), 1.27-1.31 (m, 4H),

1.63 (quint, J = 7.4 Hz, 2H), 2.35 (t, J = 7.6, 2H), 5.12 (s, 2H), 6.58 (d, J = 3.55 Hz, 1H), 7.21 (d, J = 3.55 Hz,

1H), 9.63 (s, 1H); 13C NMR (100 MHz, CDCl3) δ 14.0, 22.4, 24.6, 31.3, 34.06, 57.8, 112.6, 121.9, 152.9,

155.8, 173.3, 178.0.

8

(5-Formylfuran-2-yl)methyl benzoate (9)9

To a flame dried round-bottom flask, HMF (1.0 g, 7.9 mmol) and pyridine (4 mL) were added followed by

benzoyl chloride (1 mL, 8.7 mmol, 1.1 equiv.). The reaction mixture was allowed to stir at reflux for 1.5 h

under argon atmosphere. The mixture was quenched with cold water (50 mL), acidified with HCl 1M and

extracted with Et2O. The combined organic layers were dried over MgSO4, filtered and the solvent removed

to give the crude product that was recrystallized from Et2O/hexane as a pale orange solid (0.86 g, 47% yield). 1H NMR (300 MHz, CDCl3) δ 5.38 (s, 2H), 6.68 (d, J = 3.55 Hz, 1H), 7.24 (d, J = 3.55 Hz, 1H), 7.46 (t, J =

7.7 Hz, 2H), 7.58 (tt, J = 7.45, 1.30 Hz, 1H), 8.06 (m, 2H), 9.65 (s, 1H).

Furan-2,5-dicarbaldehyde (10)10

To a flame dried schlenk with CH2Cl2 (100 mL) at -78ºC under argon atmosphere oxalyl chloride (0.85

mL, 1.25 equiv.) was added followed by dropwise addition of DMSO (1.13 mL, 2 equiv.). After stirring for 15

minutes, a solution of HMF (1.0 g, 8.0 mmol) in CH2Cl2 (10 mL) was added dropwise and the resulting

mixture was stirred at -78ºC for an additional 30 minutes. Et3N was then added dropwise and the mixture was

allowed to warm up to RT. After 1 hour of stirring at RT, water (50 mL) was added and the resulting mixture

was washed with HCl 1M, extracted with CH2Cl2 and finally dried over Na2SO4. The resulting crude mixture

was purified by flash chromatography with Et2O/Hexane. The collected solid was further recrystallized from

hexane/EtOAc to give 305 mg (31% yield) of the product as white needles. 1H NMR (300 MHz, CDCl3) δ 7.3

(s, 2H), 9.86 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 119.4, 154.4, 179.4.

9

Furan-2-carbaldehyde (11)

The commercial available compound was purified by distillation under vacuum to give a colorless oil.

Furan-2,5-diyldimethanol (12)11

To a solution of HMF (2g, 16mmol) in dry THF (10 mL), NaBH4 (2.4g, 64 mmol) was added portionwise.

The reaction mixture was stirred at RT overnight. MeOH (7 mL) and acetic acid (9 mL) were added and

stirred at RT for 15 min then 50 mL of MeOH was added and the solvent removed under vacuum. Then twice

50 ml MeOH were added and evaporated. The mixture was dissolved in DCM/MeOH 9:1 and passed through

a pad of silica gel. The solvent was evaporated and the product was purified by silica flash chromatography

using EtOAc/Hexane to give 1.6g (80% yield) of the product. 1H NMR (300 MHz, D2O) δ 4.51 (s, 4H), 6.31

(s, 2H); 13C NMR (100 MHz, D2O) δ 55.8, 109.0, 153.6.

(5-(Ethoxymethyl)furan-2-yl)methanol (13)12

5-(ethoxymethyl)furan-2-carbaldehyde (154 mg, 1 mmol) were dissolved in MeOH (5 mL) then NaBH4

(148mg, 4 mmol) was added on 3 portions at RT. The reaction mixture was stirred for 15 min and quenched

with H2O. The MeOH was evaporated and the water phase extracted with EtOAc. The organic phase was

dried with MgSO4 and evaporated. The product was purified by silica flash chromatography using

EtOAc/Hexane to give 148mg (95% yield) of the title compound. 1H NMR (300 MHz, CDCl3) δ 1.19 (t, J =

7.0 Hz, 3H), 2.62-2.74 (b, 1H), 3.51 (q, J = 7 Hz, 2H), 4.39 (s, 2H), 6.53 (s, 2H), 6.20 (m, 1H), 6.23 (m, 1H).

10

Sodium 5-(hydroxymethyl)furan-2-carboxylate (14)13

To a closed vessel containing a solution of HMF (3 g, 24 mmol) in water (30 mL), NaOH (0.87 g, 22

mmol) was added at 0ºC, and the resulting mixture stirred for 18h at 0 ºC. The water was removed under

vacuum and ethyl acetate (2 x 50 mL) was added to the solid residue to separate the furan-2,5-diyldimethanol

(0.85g, 30% yield). From the remaining solid, the title compound was isolated by recrystallization with

ethanol (2 mL)/ethyl acetate (100 mL) to give the product as a hygroscopic solid (1.4 g, 41% yield). 1H NMR

(400MHz, D2O): δ 6.90 (d, J = 3.3Hz, 1H), 6.43 (d, J = 3.3Hz, 1H), 4.54 (s, 2H); 13C NMR (100 MHz, D2O):

δ 166.5, 155.6, 149.0, 115.7, 109.9, 55.9; ESI (-) MS: m/z 140.89.

Furan-2-ylmethanol (15)

The commercial available compound was purified by distillation under vacuum to give a colorless oil.

2,5-Bis(ethoxymethyl)furan (16)12

(5-(ethoxymethyl)furan-2-yl)methanol (390mg, 2.5mmol) was dissolved in acetonitrile (5 mL) then NaH

(0.2g, 8.3mmol) was added followed by dropwise addition of bromoethane (0.6 mL, 8 mmol). The reaction

mixture was stirred at RT overnight. The reaction was quenched with H2O and the acetonitrile was evaporated

and the water phase extracted with Et2O. The organic phase was dried with MgSO4 and evaporated. The

product was purified by silica flash chromatography using EtOAc/Hexane to give 180 mg (39%) of the

desired product. 1H NMR (300 MHz, CDCl3) δ 1.20 (t, J = 7.0 Hz, 6H), 3.52 (q, J = 7 Hz, 4H), 4.41 (s, 4H),

6.24 (s, 2H).

11

Furan-2,5-dicarboxylic acid, FDCA (17)14

To a solution of sodium hydroxide (958 mg, 23 mmol) in water (10 mL), HMF (126 mg, 1 mmol) was

added at 20 ºC, followed by the addition of potassium permanganate (363 mg, 2.3 mmol). After 10 minutes

stirring at 20ºC the precipitate was filtered off and a concentrated HCl solution was added to the filtrate until

pH 1. The resulted precipitate was separated by filtration, washed with water and dried under vacuum to give

FDCA (106 mg, 68% yield) as a white powder. 1H NMR (300 MHz, D2O) δ 7.25 (s).

2,5-Dimethylfuran (18)

The commercial available source was used as received.

2-Methylfuran (19)


(5,5'-(Oxybis(methylene))bis(furan-5,2-diyl))dimethanol (20)15

The title compound was isolated by silica gel column chromatography EtOAc/Hexane as a 10% side

product from the following reaction. To 9.1g of Et4NBr (1% water content w/w) was mixed with 2g of

fructose and 0.2g of smashed Amberlyst-15® (10% w/w). The mixture was placed at 80 ˚C and heated up to

100 ˚C for 10 min. Then the mixture was stirred at 100 ˚C for 30 min. The mixture was cooled down to RT

and the resulting solid was washed with EtOAc (50 mL). The solvent was decanted and the solid was

12

dissolved in hot EtOH (2 mL) then under vigorous stirring EtOAc (200 mL) was added. The resulting

precipitated was filtered out and the combined solutions were filtered through a pad of silica gel (10g) and

evaporated to give brown liquid of HMF (1.25g, 71%) in 88% purity by HPLC and 10% of the desire

compound which was isolated by silica flash chromatography, EtOAc/Hexane. 1H NMR (300 MHz, CDCl3) δ

4.63 (s, 4H), 6.57 (d, J = 3.55 Hz, 2H), 7.21 (d, J = 3.55 Hz, 2H), 9.63 (s, 2H); 13C NMR (100 MHz, CDCl3) δ

64.8, 112.0, 122.0, 152.9, 157.4, 177.9.

(5,5'-((1E,1'E)-hydrazine-1,2-diylidenebis(methanylylidene))bis(furan-5,2-diyl))dimethanol (21)16

HMF (1.0 g, 8.0 mmol) and hydrazine monohydrate (0.4 mL, 8.0 mmol) was dissolved in absolute EtOH

(15 mL) then 3 drops of concentrated H2SO4. The mixture was stirred at RT for 1h. The resulting solid was

filtered and washed with absolute EtOH to give the desired product 1.7 g (86% yield). 1H NMR (300 MHz,

DMSO-d6) δ 4.45 (s, 2H), 4.47 (s, 2H), 5.43 (t, 2H), 6.50 (s, 2H), 7.02 (s, 2H), 8.44 (s, 2H); 13C NMR (100

MHz, DMSO-d6) δ 55.8, 109.6, 118.4, 148.4, 150.2, 159.4.

4-Oxopentanoic acid, levulinic acid (22)


13

HPLC

Figure 1. HPLC chromatogram of 1.


14



15



16



17



18



19



20



21


22

GC-MS

Figure 18. GC-MS chromatogram and MS of major signal of 1.


5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.00.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5(x100,000)

TIC

40 50 60 70 80 90 100 110 1200.0

25.0

50.0

75.0

100.0

%

41

97695153

1268144

10966 9862

5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0

0.00

0.25

0.50

0.75

1.00

1.25

(x10,000,000)TIC

40 50 60 70 80 90 100 110 120 130 140 1500.0

25.0

50.0

75.0

100.0

%

109

53144

81

146110

8749 7942 61 12411773 95

23


5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.00.0

1.0

2.0

3.0

4.0

5.0(x10,000,000)

TIC

36.00 36.25 36.50 36.75 37.00 37.25 37.50 37.75 38.00

0.0

2.5

5.0

7.5

(x10,000,000)TIC

50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.00.0

5.0

10.0

15.0

20.0

25.0

%

18311175

61 139

53 99

15541

121

225167 195 211 239 309

24


5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0

0.00

0.25

0.50

0.75

1.00

1.25

(x1,000,000)TIC

50.0 75.0 100.0 125.0 150.00

10

20

30

40

50

%43

126

53

79

41 109

6997

1259870 16861

25



5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0

0.0

1.0

2.0

3.0

(x10,000,000)TIC

4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0

0.0

0.5

1.0

1.5

2.0

(x10,000,000)TIC

40 50 60 70 80 90 1000.0

25.0

50.0

75.0

100.0

%

96

6740 42 5149 98656154 70 7977 93

5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0-0.25

0.00

0.25

0.50

0.75

1.00(x100,000)

TIC

40 50 60 70 80 90 100 110 120 1300.0

25.0

50.0

75.0

100.0

%

41 53

51

6997

4381

109

65 12883

26


5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0

0.0

1.0

2.0

3.0

(x10,000,000)TIC

4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0

0.0

1.0

2.0

3.0

(x10,000,000)TIC

50.0 75.0 100.0 125.0 150.00.0

25.0

50.0

75.0

100.0

%

98

41

8153

69

51

7961 91 160131103 148119

27

Figure 25. GC-MS chromatogram and MS of the major signals of 18.

5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.00.25

0.50

0.75

1.00

1.25

1.50

1.75

2.00

(x100,000)TIC

40 50 60 70 80 90 100 1100.0

25.0

50.0

75.0

100.0

%

43

97

69

41 53 1125581 9884

40 50 60 70 80 90 100 1100.0

25.0

50.0

75.0

100.0

%

43

97

69112

41 5553 9870

28

Figure 26. GC-MS chromatogram and MS of the major signals of 19.

5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.00.25

0.50

0.75

1.00

1.25(x100,000)

TIC

40 50 60 70 80 90 1000.0

25.0

50.0

75.0

100.0

%

43

83 98

55

69

41 53 7081

40 50 60 70 80 90 1000.0

25.0

50.0

75.0

100.0

%

55

43 98

41

50.0 75.0 100.0 125.0 150.0 175.00.0

25.0

50.0

75.0

100.0

%

43

137

109

16281

180

91556541 119

29

NMR Spectra

Figure 27. 1H NMR spectrum of 1.

30

Figure 28. 13C NMR spectrum of 1.

31


2.22

1.07

1.00

1.13

1.87

4.54

4.79

6.42

6.43

6.90

6.90

8.41

32


55.8

6

109.

86

115.

73

148.

97

155.

60

166.

51

33


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

6.56.66.76.86.97.07.17.27.3f1 (ppm)

0

10

20

30

1

2 3

4

O56

H7

O8

9

Cl10

34


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

6.56.76.97.17.3f1 (ppm)

0

10

20

30

3.60f1 (ppm)

0

10

20

30

40

50

60

f1 (ppm)

0

50

100

150

1 2

3

O4

5

67

O8

O9

10CH311

35


-100102030405060708090110130150170190210f1 (ppm)

-30

-20

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

210

220

1 2

3

O4

5

67

O8

O9

10CH311

36


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

1

2 3

4

O56

H7

O8

9

O10

11

12

13

1415

16

17

4.554.604.65f1 (ppm)

0

50

6.56.66.76.86.97.07.17.27.37.4f1 (ppm)

0

50

100

37


0102030405060708090100110120130140150160170180190200210f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

1

2 3

4

O56

H7

O8

9

O10

11

12

13

1415

16

17

128.0128.4128.8f1 (ppm)

0

50

100

38


-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

1

2 3

4

O56

H7

O8

9

O10

Si11

12

CH313

CH314

CH315

CH316CH3

17

6.46.56.66.76.86.97.07.17.27.3f1 (ppm)

0

2

4

6

8

39


-100102030405060708090100110120130140150160170180190200210f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

1

2 3

4

O56

H7

O8

9

O10

Si11

12

CH313

CH314

CH315

CH316CH3

17

40


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

6.56.76.97.17.3f1 (ppm)

0

5

10

151

2 3

4

O56

H7

O8

9

O10

11

CH312

O13

41


-100102030405060708090110130150170190210f1 (ppm)

-4

-2

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

1

2 3

4

O56

H7

O8

9

O10

11

CH312

O13

42


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

6.76.97.1f1 (ppm)

0

10

20

30

0.00.51.01.52.02.53.0f1 (ppm)

0

10

20

30

40

50

60

701

2 3

4

O56

H7

O8

9

O10

11

12

O13

14

1516

CH317

43


0102030405060708090100110120130140150160170180190200210f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

1

2 3

4

O56

H7

O8

9

O10

11

12

O13

14

1516

CH317

44


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

2.09

1.01

1.01

2.08

1.04

2.03

1.00

1.19

1.21

1.22

2.11

3.45

3.47

3.48

5.38

6.67

6.68

7.23

7.24

7.45

7.47

8.04

8.05

8.05

8.07

8.07

9.65

1

2 3

4

O56

H7

O8

9

O10

11

12

O13

14

1516

17

18

6.46.66.87.07.27.47.67.88.08.2f1 (ppm)

0

10

20

30

45


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

12

3

O4

56 7

H8

O9

H10

O11

46


-100102030405060708090110130150170190210f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

12

3

O4

56 7

H8

O9

H10

O11

47


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

1

2 3

4

O56

OH7

8

OH9

48


0102030405060708090100110120130140150160170180190200f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

1

2 3

4

O56

OH7

8

OH9

49


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.0f1 (ppm)

-100

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

6.26.3f1 (ppm)

0

200

400

600

3.53.6f1 (ppm)

0

100

200

300

400

500

1.20f1 (ppm)

0

500

10001

O2

3

4 5

6

OH7

8

O9

10

CH311

50


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

1

O2

3

4 5

6

O7

8

CH39

10

O11

12

CH313

51


0123456789101112131415f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

1

2 3

4

O56

OH7

O8

9

OH10

O11

52


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

6.56.66.76.86.97.07.17.27.3f1 (ppm)

0

10

20

30 12

3O4

56

O7

8 9

1011

12

O1314

15

O16

O17

53


-100102030405060708090110130150170190210f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

120

130

12

3O4

56

O7

8 9

1011

12

O1314

15

O16

O17

54


0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.510.5f1 (ppm)

-10

0

10

20

30

40

50

60

70

80

90

100

110

1 2

3

O4

56

OH7

8

N9

N10

11

12 13

14

O15

1617

OH18

55


-100102030405060708090110130150170190210f1 (ppm)

0

10

20

30

40

50

60

70

80

90

1 2

3

O4

56

OH7

8

N9

N10

11

12 13

14

O15

1617

OH18

56

Toxicity curves

Compound 1

1.8 2.0 2.2 2.4 2.6 2.8 3.080

90

100

110

Log10(M)

Via

bil

ity

(%)

Compound 2

1.8 2.0 2.2 2.4 2.6 2.8 3.050

60

70

80

90

100

110

Log10(M)

Via

bil

ity

(%)

57

Compound 3

1.8 2.0 2.2 2.4 2.6 2.8 3.050

60

70

80

90

100

110

Log10(M)

Via

bil

ity

(%)

Compound 4

1.8 2.0 2.2 2.4 2.6 2.8 3.060

80

100

120

140

Log10(M)

Via

bil

ity

(%)

58

Compound 5

1.8 2.0 2.2 2.4 2.6 2.8 3.050

60

70

80

90

100

Log10(M)

Via

bil

ity

(%)

Compound 6

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

59

Compound 7

1.8 2.0 2.2 2.4 2.6 2.8 3.060

70

80

90

100

110

120

Log10(M)

Via

bil

ity

(%)

Compound 8

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

60

Compound 9

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

Compound 10

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

61

Compound 11

1.8 2.0 2.2 2.4 2.6 2.8 3.070

80

90

100

110

Log10(M)

Via

bil

ity

(%)

Compound 12

1.8 2.0 2.2 2.4 2.6 2.8 3.080

90

100

110

120

130

Log10(M)

Via

bil

ity

(%)

62

Compound 13

1.8 2.0 2.2 2.4 2.6 2.8 3.080

90

100

110

120

130

140

Log10(M)

Via

bil

ity

(%)

Compound 14

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

63

Compound 15

1.8 2.0 2.2 2.4 2.6 2.8 3.070

80

90

100

110

120

Log10(M)

Via

bil

ity

(%)

Compound 16

1.8 2.0 2.2 2.4 2.6 2.8 3.060

80

100

120

140

Log10(M)

Via

bil

ity

(%)

64

Compound 17

1.8 2.0 2.2 2.4 2.6 2.8 3.090

100

110

120

130

Log10(M)

Via

bil

ity

(%)

Compound 18

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

65

Compound 19

1.8 2.0 2.2 2.4 2.6 2.8 3.080

90

100

110

Log10(M)

Via

bil

ity

(%)

Compound 20

1.8 2.0 2.2 2.4 2.6 2.8 3.00

50

100

150

Log10(M)

Via

bil

ity

(%)

66

Compound 21

1.8 2.0 2.2 2.4 2.6 2.8 3.080

90

100

110

120

130

Log10(M)

Via

bil

ity

(%)

Compound 22

1.8 2.0 2.2 2.4 2.6 2.8 3.085

90

95

100

105

110

115

Log10(M)

Via

bil

ity

(%)

67

References

(1) Shimizu, K.; Uozumi, R.; Satsuma, A. Catal. Commun. 2009, 10, 1849. (2) Svilen, P.; Coelho, J. A. S.; Afonso, C. A. M. Chemsuschem 2012, 5, 1388. (3) Monien, B. H.; Frank, H.; Seidel, A.; Glattt, H. Chem. Res. Toxicol. 2009, 22, 1123. (4) Sanda, K.; Rigal, L.; Gaset, A. Carbohydr. Res. 1989, 187, 15. (5) Mascal, M.; Nikitin, E. B. Angewandte Chemie International Edition 2008, 47, 7924. (6) Cottier, L.; Descotes, G.; Eymard, L.; Rapp, K. Synthesis 1995, 303. (7) Cottier, L.; Descotes, G.; Soro, Y. J. Carbohyd. Chem. 2005, 24, 55. (8) Dai, H. L.; Shen, Q.; Zheng, J. B.; Li, J. Y.; Wen, R.; Li, J. Lett. Org. Chem. 2011, 8, 526. (9) Bognar, R.; Herczegh, P.; Zsely, M.; Batta, G. Carbohydr. Res. 1987, 164, 465. (10) Mehdi, H.; Bodor, A.; Lantos, D.; Horváth, I. T.; De Vos, D. E.; Binnemans, K. The Journal of

Organic Chemistry 2006, 72, 517. (11) Cottier, L.; Descotes, G. R.; Soro, Y. Synthetic. Commun. 2003, 33, 4285. (12) Balakrishnan, M.; Sacia, E. R.; Bell, A. T. Green Chemistry 2012, 14, 1626. (13) Gupta, N. K.; Nishimura, S.; Takagaki, A.; Ebitani, K. Green Chemistry 2011, 13, 824. (14) KAISHA, C. K., 2007. (15) Cottier, L.; Descotes, G.; Eymard, L.; Rapp, K. Synthesis 1995, 1995, 303. (16) Knotz, F. scientia pharmaceutica 1961, 29, 20.

si tx-com-03-2014-000019.r1 proofs · 4 5-(hydroxymethyl)furfural, hmf (1)1 to a round-bottom flask...

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