National High Magnetic Field Laboratory at Florida State University
FT-ICR
Mass Spectrometry Advantages
Every molecule has mass!
Mass Defect as a Chromophore
Isotopes: Tracing/Quantitation
Attomoles
Interpretation (One Peak/Cpd)
(m/z)max(m/z)minm/z
Peak Capacity = m50%
(m/z)max - (m/z)min
m50%
Separation Method
Maximum # of Components
MaximumPeak Capacity
HP-TLC 6 25
Isocratic LC 12 100
Gradient LC 17 200
HPLC 37 1,000
CE 37 1,000
Open Tubular GC 37 1,000
ESI FT-ICR MS 525 200,000
B
h B= q B m=
BNMR or EMR ICR
Differential Amplifier
FT
100 150 200 250Frequency (kHz)
7+
8+
10+
11+
12+
9+
600 1000 1400 1800
12+
11+
10+
9+
8+
7+
m/z
0
80 240 400Time (ms)
ImageCurrent
BovineUbiquitin
10721071
mz
A= + B
2
Capillary tip
Taylor Cone
HV
Counter Electrode
Electrospray Ionization
Adapted from: Enke, C. Anal. Chem. 1997, 69, 4885-4893.
Heated Metal Capillary
Sleno et al.
178.10178.06178.04 178.08m/z
178.07233
178.08043
178.08489FT-ICR (IRMPD)
QqTOF (CID)
178.06107
C8H8N3O2+
m/z 178.06110
+ 13C1
12C713C1H9N4O+
m/z 178.08044
N
N
NHHO
O
N
N
NH
OH2N
C8H10N4O+•
m/z 178.08491
+N
O
C
N
H2N
N
C
CH2
C7H8N5O+
m/z 178.07234
N
N OH2N
N
NH2N
HO
N
NHO
N
N
m/z 220
-0.04
-0.03
-0.02
-0.01
0
0.01
0.02
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34
1H
2H13C 14N 15N
16O
31P32S 34S
12C
AtomicMass Defects
(Dalton)
0
2
4
6
8
10
521.0 521.1 521.2 521.3 521.4 521.5
(1 mDa Bins)
CcHhNnSsOo
S. KimRodgers
Mass
0
2
4
6
8
10
521.10 521.105 521.110 521.115 521.120
(1 mDa Bins) S. KimRodgers
Mass
CcHhNnSsOo
0
1
2
3
4
5
6
7
521.100 521.105 521.110 521.115 521.120
(0.5 mDa Bins)
Mass
S. KimRodgers
CcHhNnSsOo
0
1
2
3
4
521.110 521.112 521.114 521.116 521.118 521.120
(0.1 mDa Bins)
CcHhNnSsO
o
Mass
+/- 100 ppb =1 Composition per bin!
Negative-Ion APPI FT-ICR MS
-2 -1 0 1 2
100
90
80
70
60
50
40
30
20
10
Mass Accuracy vs. Relative Abundance (12,449 Assigned Masses)
Mass Accuracy (ppm)
Re
lati
ve
Ab
un
dan
ce
S. American Crude Oil
± 100 ppb
229225221
C17H21+
C16H33+
C16H17O+
C15H13S+
Measured Theoretical225.07326 225.07325 225.12733 225.12739 225.16375 225.16378 225.25769 225.25768
m/z 250200150 300
C15H13S+
C16H17O+
C17H21+
C16H33+
Raw Diesel Feedstock 1L Septum Injection
RodgersAndersenWhiteHendrickson
C8H7N
Isomers
N- H+
H N
+ H+
NH
+N
900800700600500400300200
~
-900 -800 -700 -600 -500 -400 -300 -200m/z
17,000+ Compositionally Distinct Components Resolved by High Resolution 9.4 Tesla Electrospray FT-ICR MS
6,118 resolved components
11,127 resolved components
0
Positive Ion ESI MSNegative Ion ESI MS
HugheyHendricksonRodgersQian
m/z 900800700600500400300
11,000 Peaks250 < m/z < 900
m/z492.475492.400492.325492.250
Expansionat m/z 492C3 SH4
Positive ESI FT-ICR MSS. Amer. Heavy Crude Oil
m2 – m1 = 3.4 mDa
HugheyHendricksonRodgersQian
m/z431.4431.3431.2431.1431.0
8 σ
m/z700640580520460400340280
*
63 Spectral Peaks above 8 σ62 Spectral Peaks Assigned* Not assigned
APPI FT-ICR MS
Incr
ea
sin
g
DB
E
Ke
nd
ric
k M
ass
De
fect
Incr
ea
sin
g
DB
E
0.1
0.2
0.3
0.4
0.5
0.0
0.1
0.2
0.3
0.0
0.4
0.5
300 400 500 600 700 800 900Nominal Kendrick Mass0.00 0.03
0.00 0.07
Crude Oil B
Asphaltenes Deposit B
m/z650550450350250
Canola Oil
Olive Oil
Soybean Oil
Negative-Ion ESI FT-ICR MS
WuRodgers
m/z285284283282281280279278277
C18:3
C18:2
C18:1
C18:0
C18:2
C18:1
C18:0
C18:3
C18:2
C18:1C18:0
Canola Oil
Olive Oil
Soybean Oil
Fatty Acids
WuRodgers
m/z890885880875
Canola Oil
Olive Oil
Soybean Oil
TriacylglycerolsC54:6
C54:5
C54:8
C54:7WuRodgers
m/z455.5455.4455.3455.2455.1
C30H47O3-
C29H43O4-
C25H43O7-
C24H39O8-
C27H35O6-
C23H35O9-
C26H31O7-
C22H31O10-
C22H32O8P-
C25H27O8-
C21H28O9P-
Negative-Ion ESI of Olive Oil
WuRodgers
0
4
8
12
16
0
4
8
12
O2
Canola Oil
Olive Oil
O3 O4 O6O5 O7 O9O8 O10
0
20
40
60
S NP
Soybean Oil
Acidic Species Detected by Negative-Ion ESI16
WuRodgers
m/z
435430425420415410405400
C27H45O2-
(δ-tocopherol)
C28H47O2-
(β, γ-tocopherol)
m/z429.4429.3429.2429.1
C29H49O2-
(α-tocopherol)
Tocopherols in Soybean Oil
O
R1
OH
R2
R3
Negative-Ion ESIWuRodgers
m/z415.6415.5415.4415.3415.2415.1
C28H47O2- (β, γ-tocopherol)
Canola Oil
Olive Oil
Soybean Oil
C19H27O10- C20H31O9
-
WuRodgers
m/z657.62657.58657.54657.50
C41H70O4P+
C41H69O6+
C45H69O3+
C42H73O5+
C46H73O2+
C43H77O4+
C47H77O+
Positive-Ion ESI FT-ICR MS
Soybean Oil
WuRodgers
m/z610605600
Canola Oil
Olive Oil
Soybean Oil
Diacylglycerols
C36:3C36:2
C36:1
C36:4
C36:5
WuRodgers
274
278
282
20
40
60
80
100
PureOlive
PureSoybean
C18:3
Fatty Acids
Relative Abundance %
C18:2
WuRodgers
876
880
884
20
40
60
80
100
PureOlive Oil
PureSoybean Oil
Triacylglycerols
C54:7
C54:5C54:6
Relative Abundance %
WuRodgers
NHO
OH
OOOH
OH
OH
OH
OOH
NHAc
OH
O
OH
OH
O
OH
OH
OH
ONHAc
OH
OH COOHOH
OH O
O
OO
GM1a
GM1b
NHO
OH
O
O
OOH
OH
OH
OOH
NHAc
OH
O
OH
OH
O
OH
OH
OHONHAc
OH
OH COOHOH
OH
O
OO
OH
HexNAc = N-Acetylhexosamine
NeuAc = N-Acetylneuraminic Acid
(Sialic Acid)
Hex = Hexose
200 400 800 1000 1200 1400600
Ceramide
Ceramide’ [M+2H] 2+
’’
’
’ )M-( ’)M-(
) M-(
’)M-( M-
’)M-(
)M-(
m/z
’(fr
om
0,2)
oo o o
O
OH
HNO
[M+2H]2+
’
’
’)M-(
’)M-(
)M-(’)M-(
M-( )’
200 ms IRMPD of GM1
100 ms IRMPD of GM1
HNO
OH
O
o o oo
200 400 800 1000 1200 1400600
[M-2H]2-
Cer
amid
e’
Cer
amid
eCer
amid
e’’
~1.5~
’
Sp
hin
go
sin
e-N
’ ’)M
-(
)M
-(
’)M
-(
’
’)M
-(
’)M
-(
M-
)’M
-(
m/z
oo o o
HNO
OH
OHNO
OH
O
o o oo’(
fro
m
0
,2)
’
ECD of GM1
Sugar
Lipid
M-x
Unknown
Amino Acid
0
0.2
0.4
0.6
0.8
1
0 500 1000 1500
Neutral Mass
and/or
HNO
OH
O
MassDefect
Sialic Acid
Hexose
HexNAcHN
O
OH
O
Hydrocarbon
Peptides fromDigest andStandards
McFarland
Atmospheric Pressure PhotoionizationFT-ICR MS
http://www.chem.agilent.com
Inlet
Nebulizing GasNebulizer
Vaporizer
Drying Gas
Capillary
UV Lamp
h To FT-ICR MS
M
+ H+
- e-
[M+H]+
M+•
Nominal Mass
Odd Even
N0, N2, N4, … [M+H]+ M+•
N1, N3, N5, … M+• [M+H]+
Nitrogen RuleMcLafferty, Interpretation of Mass Spectra, 1993
APPI
[C30H41N1S1 + H]+
[C32H37N113C1]+ •
[C33H37N1 + H]+
m/z
448.310448.305448.300448.295448.290
4.5 mDa 3.4 mDa
Why Ultrahigh Mass Resolving Power?
South American Crude Oil3.4 mDa C3 versus SH4
4.5 mDa 13C versus CH
PurcellRodgers
[M+H]+
M+•
0
Ion Energy
Ion Trapping Period
Upper Mass LimitNumber of Ions
Mass Resolving Power
Scan Speed (LC/MS)
Highest Non-Coalesced Mass
0
7 T
7 T 9.4 T9.4 T
21 T
B (tesla)2121
B (tesla)
14.5 T
14.5 T
21 T
(2004)
(1995)(1985)
#2764 IT: 119.142 ST: 6.31 uS: 1 CS: 2 AMW: 1569.67 NL: 6.55E4F: FTMS + p ESI Full m s [ 0.00-6144.00]
0 500 1000 1500 2000 2500 3000 3500 4000 4500 5000 5500 6000
m /z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Relat
ive In
tensity
68.7004390.1465
772.3228 1192.9299 1624.7856 2084.4141 2499.5969 3175.2803 3667.2979 4082.1877 4361.0347 4825.6611 5262.9180 5699.0908
Glu fib 500K FT_041001162804 #1 RT: 0.00 AV: 1 NL: 2.98E5T: FTMS + p ESI Full ms [ 400.00-2000.00]
783 784 785 786 787 788 789 790m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lativ
e A
bu
nd
an
ce
0 2 4 6
Time (s)
783 784 785 786 787 788 789 790m/z
Glu-Fib, 1 pmol/μL
785.842
786.343
786.845
HendricksonMackayQuinn
m/m50% = 914,000
External Calibration
Mass Accuracy: 50 ppb
Nanomate LTQ Broadband FT ICR MS
857 858m/z
Absorption
Magnitude Ubiquitin
BeuBlakneyHendricksonQuinn
[M+10H]10+
0
Ion Energy
Ion Trapping Period
Upper Mass LimitNumber of Ions
Mass Resolving Power
Scan Speed (LC/MS)
Highest Non-Coalesced Mass
0
7 T
7 T 9.4 T9.4 T
B (tesla)21
B (tesla)
14.5 T (2004)
(1995)(1985)
$1.5 M
$0.5 M$0.2 M
$?? M
NHMFLKBSIPNNL
21 T
(2009)
NSF
Florida State U.
NIH ExxonMobil
STINT (Sweden)
Schlumberger-Doll
Dow
Support for ICR at NHMFL
Fluor Canada
Saudi Aramco
ThermoElectronOil Phase DBR
Baker Petrolite
UnoCal
KBSI (Korea)
0
100
200
300
400
500
600
700
1976 1980 1984 1988 1992 20001996 2004
WorldwideFT-ICR MS Systems
NHMFL ICRProgram Starts
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