Mass Spectrometry Facility
Advanced Analysis Centre
University of Guelph
Presented by:
Dr. Dyanne Brewer
Mass Spectrometry Facility Manager
May 2014
MS Sample Submission Workflow
Sample
Submission
Sample
Preparation
Data
Acquisition
Data
Interpretation
Submission Forms:
http://www.uoguelph.ca
/~bmsf/
Training Available:
GC-MS derivitization
Protein digestion
Training Available:
GC-MS
LC-MSn
Training Available:
Metabolomics software
Proteomics software
Instrumentation
Agilent GC-MS
Bruker GC-MS-MS
Bruker LC-MSn
Solid, liquid, gas, SPME
GC- gas chromatography
LC- liquid chromatography
MS- mass spectrometry
Agilent LC-HR
MS-MS
Sample Type: Small Molecule
MS confirmation
Volatile/semivolatile
GC
separation
MS-MS
quantitation
Small
Molecule
Non-volatile
LC
Separation
MS
confirmation/formula
MS-MS characterize/quantitate
Small Molecule GC-MS
Sample Collection
Metabolite Extraction
Compound Derivitization
8.744 min, Scan: 1089
Search
TIC; KIT_051964_T120_4-4-2013_Metabol2.xms; Filtered
100%
174.2
5.251e+8
73.1
4.742e+8
10.0
75%
50%
147.1
1.494e+8
7.5
25%
248.2
1.042e+8
59.6
7.291e+7
GCps
0%
Match
5.0
Match 1 of 12
174.0
999
100%
Si
75%
Si
N
O
2.5
50%
73.0
510
O
Si
25%
86.0
132
0.0
147.0
208
248.0
163
0%
7.5
10.0
12.5
15.0
17.5
20.0
minutes
Gas Chromatogram
100
R.Match: 845, F.Match: 819
200
300
400
500
600
m/z
NIST match to Glycine
N,N-bis(trimethylsilyl) trimethyl ester
Dr. Mutch UoG HHNS
Small Molecule Mass Accuracy
Best
TRUE
ID Source
MFG
Formula
C49 H54 Cl5 N8 O9 P
Species
(M+H)+
m/z
1105.2269
Score
94.53
Diff (ppm)
-0.33
Score (MFG)
94.53
Dr. Manderville UoG Chem
Small Molecule LC-MS
Intens.
x108
2.5
2.0
1.5
1.0
0.5
0.0
0
2
4
6
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 271.0 +All MS2
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 727.0 +All MS
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 303.0 +All MS2
Cherry-profile-Jan-21-2014_BD2_01_1145.d: Base Peak UV Chromatogram, 520 nm
8
10
12
14
16
Time [min]
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 449.0 +All MS2
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 301.0 +All MS2
Cherry-profile-Jan-21-2014_BD2_01_1145.d: EIC 317.0 +All MS2
UV
Pelargonidin
Cyanidin
Peonidin
Delphinidn
Malvidin
Petunidin
520 nm
271
287
301
303
331
317
Intens.
[%]
100
+MS2(609.4), 10.2min #571
301.0
-peonidin
80
-hexose
60
40
463.0
20
-deoxihexose
0
200
400
600
800
1000
1200
1400
m/z
Dr. Kirakosyan Un. Michigan
Small Molecule LC-MS
Heart
Bladder
kidney
Brain
Tissue Extract
Calibration Curve of Peonidin 3-O-glucoside
3000000
y = 1017.4x + 20978
2000000 R² = 0.99
sample name
1000000
0
0
2000
4000
RT [min] Area
concentration_peonidin dihexoside_fg/ul
1%heart_1.54g_(5)
10.2
42548
32.6
1%Bladder_0.75g_(1)
10.2
12946
3.2
1%kidney_1.4g_(14)
10.2
91933
81.7
1%brain_0.94g-(9)
10.2
14337
4.6
Dr. Kirakosyan Un. Michigan
Metabolomics Workflow
Separate and
Detect MS
GC-MS/LC-MS
Compare
and Stats
MS database
search targets
MassHunter/MPP
software
In-house
Metlin/NIST
library or online
Target MS/MS
Compare to
database/standard
ID
GC-MS data
Pathway?
Sample Type: Large Biomolecule
Intact
characterization
Protein
Clean-up LC
MS
confirmation
Solution
digest
Identification
LC
In-gel digest
Auto MS-MS
Database
search/de
novo
sequencing
PTM id
Protein MW
Protein solution
Deconvoluted data
Clean-up LC
Raw data
27,310.4656 amu
Dr. Brown UoG Physics
Protein Identification
Protein digest
LC-Chromatogram
Protein
Group Protein ID Accession -10lgP
Colicin_E1
1
6181
_Q372C 283.51
Coverage
(%)
#Peptides #Unique
69
60
60
PTM
N
Descriptio
n
Colicin_E1
21734 _Q372C
Avg. Mass
Peptide Fragmentation
Protein Identification
Dr. Merrill UoG MCB
Mass Spectrometry Facility
Advanced Analysis Centre
University of Guelph
Manager: Dr. Dyanne Brewer
Research Associate: Dr. Armen Charchoglyan
Location: Science Complex Rm 1205
Email: dbrewer@uoguelph.ca
Phone: x58649 lab x53793 DB office