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Tandem Mass Spectrometry: An arrangement of two or more mass spectrometers (tandem-in-space) or mass spectrometry experiments (tandem-in-time) placed one behind the other. Acronym: MS/MS; MS3; MSn
ionization source A+ , B+ , C+ , D+
precursor ion
A+
A1+ A+ m/z
mass analyzer II
isobar
distinct fragments
OH CH2 N
O C C C NH NH2
+
Precursor Ion
N C NH2 NH
C C
CH N CH2CH2O C NH2 N
CH
Product Ion
m/z
O
196
OH CH2 CH N NH3 O CH2 C C C NH N CH2 CH
m/z
CH2 OH
152
O CH2
C N
+
OH CH2 N
O C C NH CH
C C NH
NH
+C
N C NH C C
C NH3
+C
CH N CH2CH2O C NH
+C
m/z
O
196
O CH2 C N CH C NH2
+
m/z 135
CH2 OH CH2 OH CH2 CH C NH2 N NCNH2 NH2 C N O CH2 OH CH2 CH CH2CH2O NH2 O C C NH CH
C N C NH2 NH2 C C
+C
C
+C
m/z
O
196
m/z 110
OH CH2 CH
CH2 C N C NH2 NH C C NH
+
+ CH2
CH2
OH
m/z
196
m/z
45
LC-MS/MS
LC-MS
128 fmol
1.02 pmol
LC-MS/MS
LC-MS
128 fmol
1.02 pmol
Full MS scan
Mass analyzer
scan
select ion
scan
x10
High Resolution!
x10
It was realized that MALDI ions are not as soft as many had expected. First commercial mass spectrometer that performed PSD analysis was introduced ~ 1994.
ESI
ground voltage
Shielding is important for TOF chamber to create a field-free region floated at high potential
(From Chernushevich et al., JMS, 2001, 36, 849)
Pulsing too fast: overlap with slowest ion from previous pulse Pulsing too slow: low duty cycle Typical pulsing rate is 5-10 kHz with 5-30% duty cycle
(From Chernushevich et al., JMS, 2001, 36, 849)
Extraction Pulsing Rate, Duty Cycle, and Gating pulse for Duty Cycle Enhancement @ a specific m/z region
Using gating pulse, ~ 100% duty cycle can be achieved @ a specific m/z region, At the expense of low duty cycles in other m/z regions
Can Q-TOF be used for various MS/MS modes as provided by triple quadrupole? (yes) How about efficiencies? (depends)
Product ion scan
select ion
(CID)
scan
scan
(CID)
scan
scan
(CID)
select ion
select ion
(CID)
select ion
mpre+
Scan
Product ion scan Precursor ion scan Neutral loss scan Selected reaction monitorg
mpro+ + mneu
mpro+
Vary Fix Vary Fix
Efficiency
mneu
Vary Q-TOF >> TSQ Vary Fix Fix
mpre+
Fix Vary Vary Fix
Q-TOF ~ TSQ
Gating pulse for Duty Cycle Enhancement @ a specific product ion is useful in precursor ion scan
Using gating pulse, ~ 100% duty cycle can be achieved @ a specific product ion
Liquid Chromatography
Exact mass measurement of precursor and product ions for structural confirmation and identification by Q-TOF
Exact mass measurement of precursor and product ions for structural confirmation and identification by Q-TOF
Fragmentation
An example of product ion data of tryptic peptides generated by tandem time-offlight (TOF/TOF) mass spectrometry