Standard mixture is composed of: 2-methyl furan, Furfuryl Alcohol, Shikimic Acid, Vanillic Acid, P-Cresol, 3,5-dimethoxy toluene, Eugenol, 5-

(hydroxymethel) furfural, Syringol, Apocynin, Levoglucosan, Syringaldehyde, Acetosyringone, Coniferyl Aldehyde, Pyrene

• Pyrolysis of lignocellulosic biomass has shown promise as an alternative energy source to fossil fuels.

• Comprehensive characterization of biomass pyrolyzates is critical for producing high quality biofuels.

• Traditional GC-EI-MS has many limitations, especially in identifying the compounds that are not in the database.

• Dopant assisted atmospheric pressure chemical ionization (dAPCI) coupled to GC-TOFMS is a complementary tool; however, structural information is limited.

Exploring Structural Information using In-source CID on Micropyrolyzer-GC-dAPCI-TOF Mass Spectrometry

Nathan R. Bond1; Daniel P. Cole1; Carolyn P. Hutchinson1; Young Jin Lee1 1 Department of Chemistry, Iowa State University, Ames, IA

Overview Results

• To explore the use of in-source CID for structural analysis of biomass pyrolyzates in Py-GC-dAPCI-TOFMS.

Instrumentation

Materials • Standards are obtained from Sigma Aldrich (see first chromatograph) • Standard mix is prepared at 1 mM per compound with 5 μL injection. Methods • Red oak pyrolysis: 500°C on Frontier micropyrolyzer. • GC programing: Oven temp. held at 35° for 5 min, increased at

4° per min to 260°, and held for 5 min. • Standard condition: Fragmenter -95V, Skimmer -65V • Fragmentation conditions: Fragmenter -160V, Skimmer -145V

Experimental

Conclusions

Autosampler or micropyrolyzer

TOF MS Agilent 6200

series

Ammonia gas

APCI interface

GC

DB-1701 column (J& W, 30 m x 0.25 mm x 0.25 μm)

• Understanding MS/MS of standard compounds allows to interpret fragmentation pattern of unknowns.

• Most ammonium adducts are resulted in the loss of ion signals in fragmentation condition. Thus, it is not effective for levoglucosan or other cellulose pyrolysis products.

Acknowledgements Future Direction

• Test tentative identifications against standards • Run replicate experiment • Study the effect of changing other parameters • Examine other standard compounds to observe

fragmentation patterns

6x10

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+APCI TIC Scan Frag=95.0V 01_1mM std mix_5ul ALS_GCsep.d

1

Counts vs. Acquisition Time (min)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56

Acetosyringone (RT:46.5 min) Syringaldehyde (RT:44.7 min)

3,5-dimethoxytoluene (RT:29.3 min) Coniferyl Aldehyde (RT:47.35 min)

Unknown at 44.8 min.(Syringaldehyde) Unknown at 40.5 min (3,5-dimethoxyphenone)

Unknown at 32.4 min (4-hydroxy-3-methoxystyrene) Unknown at 36.4 min (2-methoxy-4-propenylphenol)

5x10

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+APCI TIC Scan Frag=160.0V 02_1mM std mix_5ul ALS_GCsep_frag160V_skimmer145V.d

1

Counts vs. Acquisition Time (min)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56

197.0756

213.0944

207.0301

190 195 200 205 210 215 220

183.0608

199.0793

208.0378175.0495

4

160 165 170 175 180 185 190 195 200 205 210 215 220 225

4x10

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1.

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2.

+APCI Scan (46.291-46.658 min, 520 Scans) Frag=160.0V 02_1mM std mix_5ul ALS_GCsep_frag160V_skimmer145V.d Subtract

181.0480

196.0715140.0459

155.0689125.0224108.0200

167 0322

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

100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200

6x10

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+APCI TIC Scan Frag=95.0V 04_py_3rods_redoak_nonacid_infused_GC sep_frag95V_skimmer65V.d

1

Counts vs. Acquisition Time (min)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54

6x10

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+APCI TIC Scan Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_frag160V_skimmer145V.d

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Counts vs. Acquisition Time (min)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54

Micropyrolyzer-GC-dAPCI-

TOFMS

CID

153.0874

0980 160 1653

45 150 155 160

3x10

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3.

3.

+APCI Scan (29.241-29.365 min, 176 Scans) Frag=160.0V 02_1mM std mix_5ul ALS_GCsep_frag160V_skimmer145V.d Subtract (2)

153.0894

138.0661123.0789

109.0637

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

85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165

179.0656

195.0844

213170 0766

170 175 180 185 190 195 200 205 210 2

3x10

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+APCI Scan (46.861-47.762 min, 1274 Scans) Frag=160.0V 02_1mM std mix_5ul ALS_GCsep_frag160V_skimmer145V.d Subtract

179.0689

147.0429

161.0584119.0484

107.0487 135.0431

124.0508

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

85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190

180.0758

198.1097

167 0658 208 0345

170 175 180 185 190 195 200 205 210

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+APCI Scan (40.412-40.674 min, 372 Scans) Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_frag160V_skimmer145V.d Subtract

180.0757

165.0525

137.0580

121.0636

149.0580105.0439

115.0383 197.0769129 0508 207 0974

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

80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200 205 210

151.0732

158.0793

146.0791

45 150 155 160

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+APCI Scan (32.281-32.478 min, 280 Scans) Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_frag160V_skimmer145V.d Subtract

151.0735

135.0428

107.0485

119.0482

140.0452

166.0443115.0384

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

80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180

NRB acknowledges Alice & Donald Hudson Scholarship to support undergraduate research and Professor Emeritus Robert Angelici to support the travel to attend the 2014 American Society for Mass Spectrometry Conference.

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+APCI Scan (44.611-44.886 min, 390 Scans) Frag=160.0V 02_1mM std mix_5ul ALS_GCsep_frag160V_skimmer145V.d Subtract

182.0557

123.0431

167.0324140.0453155.0687

111.0434

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

100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190 195 200

CID CID

CID

CID

CID CID

[MH]+

[M.NH3]+.

[MH]+

[MH]+

[MH]+

~

-C3H2O

-C4H8O -C4H8O2

-C2H4O

-C2H2O -C3H2O

M+.

M+.

~

[MH]+ [MH]+

-H2O

150.0661

-C3H4O2

-C3H2O

-CHO

C9H10O2

-C2H4O -CH4

-C2H3O2

-CH4O

-CH3O

-C2H3O -CH3

C10H12O3

Purpose

Overview

GC-TIC Chromatograms MS and MS/MS spectra of selected compounds

Standard mix without fragmentation

Standard mix with fragmentation

Pyrolysis of red oak without fragmentation

Pyrolysis of red oak with fragmentation

M+.

[MNH4]+

[MNH4]+

[M.NH3]+. M+.

[MNH4]+

[M.NH3]+.

M+.

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+APCI Scan (44.727-44.940 min, 301 Scans) Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_frag160V_skimmer145V.d Subtract

182.0538

105.0442 123.0429

140.0451155.0683 167.0316

111.0431

131.0476 151.0730115.0389 163.0727

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

80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170 175 180 185 190

183.0617

200.0881

9 0471

70 175 180 185 190 195 200 205 21

M+.

M+.

[MH]+

[M.NH3]+.

[MNH4]+

M+.

M+.

[MH]+

-C2H2O -C3H2O

M+.

[MH]+

-C2H4O2

-CH4 -CO

Acetosyringone

Syringaldehyde

Coniferyl Aldehyde

3,5-dimethoxytoluene

kimmer65V.d Subtract

164.0811

182.1151

0738 172.0945188 0

155 160 165 170 175 180 185 19

4x10

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+APCI Scan (36.295-36.429 min, 190 Scans) Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_frag160V_skimmer145V.d Subtract

164.0814

149.0580105.0675

137.0583

133.0634121.0637

115.0390

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

102 104 106 108 110 112 114 116 118 120 122 124 126 128 130 132 134 136 138 140 142 144 146 148 150 152 154 156 158 160 162 164 166 168

M+.

[MH]+

[MNH4]+

-CH4

-CH4O

-C2H4 -C2H4O2

-C2H4O

M+.

M+.

C10H12O2

44.8 36.4 40.5

32.4

3

Pyrene

P-Cresol

Levoglucosan

1

9, 10 2

4

Compounds listed numerically: Lactic Acid, 2-Butonic Acid, 2,5-Dimethyl-3-methylene P-dioxane, 3-Oxobutanoic Acid, 2-(5H) Furanone, a-Methyl furan, Furfural, p-/o-dioxanone, 2-propenoic Acid, Ethyl ester, 5-Methyl 2(3H)-furanone, Vanillic Acid, Levoglucosane Compounds listed numerically: 5-methoxy-2-furoic Acid, Maltol, Syringol, Vanillyl methylcetone, Ethyl vanillylether, Acetosyringone, Phenol, Methyl 10-hydroxydec-trans-8-ene-4,6-diynoate

12

5

6 11

7,8

13

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5