1. 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.
AcknowledgementsFuture 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
0
0.
0.
0.
0.
0.
0.
0.
0.
0.
1
+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
0
1
1
2
2
3
3
4
4
5
5
+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
0
0.
0.
0.
0.
0.
0.
0.
0.
0.
1
1.
1.
1.
1.
1.
1.
1.
1.
1.
2
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
155.0689
125.0224108.0200
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
0
0
1
1
2
2
3
3
4
4
5
5
6
6
7
7
8
+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
0
0
0
0
1
1
1
1
2
2
2
2
3
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3
3
4
4
4
+APCI TIC Scan Frag=160.0V 05_py_3rods_redoak_nonacid_infused_GC sep_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
Micropyrolyzer-GC-dAPCI-
TOFMS
CID
153.0874
0980 160 1653
45 150 155 160
3x10
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0.
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0.
1
1.
1.
1.
1.
2
2.
2.
2.
2.
3
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
138.0661
123.0789
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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1
1
1
1
1
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2
2
2
2
+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
161.0584
119.0484
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
4x10
0
0.
1
1.
2
2.
3
3.
4
4.
+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
4x10
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0
0
0
0
1
1
1
1
1
2
+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.
3x10
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1.
1.
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1.
2
2.
2.
+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
167.0324140.0453
155.0687
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
CIDCID
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+.
4x10
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1
1
1
1
1
2
2
+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
140.0451
155.0683 167.0316
111.0431
131.0476 151.0730
115.0389 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 172.0945
188 0
155 160 165 170 175 180 185 19
4x10
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1
1
1
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2
2
2
2
+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
149.0580
105.0675
137.0583
133.0634
121.0637
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
1
1
3
3
2
2
4
6
4
7
7
6
8
8
5
5