The Most Powerful Instrument
DESIGNED TO ADVANCE YOUR SCIENTIFIC PURSUITS
Tribrid Mass Spectrometer
PUSHING NEW
FRONTIERS IN SCIENCE
Breakthroughs in science rely on great scientists
making rapid progress answering challenging questions.
“How does lipid metabolism affect cancer?”
“How do non-cancerous cells mutate?”
“What protein modifications are indicative of a disease state?”
As a leading scientist, you need to be equipped with
the latest innovations in analytical technology.
With the Thermo Scientific™ Orbitrap Fusion™
Lumos™ Tribrid™ MS, we are committed to
keeping pace with your scientific pursuits
through continuous innovations.
The Most Powerful Instrument
Designed for High Impact Discoveries
The Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS delivers superior sensitivity, selectivity and
versatility to enable life scientists to obtain the highest quality data. It is designed to pursue the most difficult
analyses, including multiplexed quantitation of low-abundance peptides in complex matrices, characterization of
positional isoforms of intact proteins, resolution of isobaric metabolites, protein structure characterization using
chemical crosslinking and the deepest mining of challenging post-translational modifications.
You trusted us to make this most versatile and powerful mass spectrometer a reality.
We now trust you with groundbreaking science and high impact discoveries.
Multiplexing Targeted Analysis
Accurately Quantifying Hundreds of Proteins in 90 Minutes
Multiplexed analyses using isobaric mass tags are widely utilized for high throughput quantitative comparisons of protein abundances.
The TMT SPS MS3 workflow available on the Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS is a proven method which
enables the simultaneous analysis of 10 samples together with improved quantitative accuracy. Pushing the multiplexing frontiers even
further, targeted assays incorporating this technique can now be built to detect and quantify very low levels of target peptides even from
undetected precursors. This new TMT SPS tMS3 workflow further benefits from the enhanced sensitivity of the Orbitrap Fusion Lumos MS,
which boosts the number of quantifiable peptides present at low levels.
List of
Peptide
Targets
Synthetic peptide targets
labeled with TMT0
mixed
with
Native peptides from digested
samples labeled with TMT10
MS DETECTION
Synthetic
TMT0-labeled
Target
8.81
6.59
12.26
Isolation
Offset
15.14
27.32
30.33
36.50
50.98
46.44
77.93
41.19
10
TMT10-labeled
Native Peptide
Target
20
30
40
61.67
50
Time (min)
60
67.73
70.73 75.15
70
81.96
80
Base peak Extracted Ion Chromatogram (XIC). Purple peaks are XICs of TMT0-labeled peptides.
Everley et al. WP 663, ASMS 2016.
m/z
The synthetic TMT0-labeled peptide is
detected in the full MS scan. The native
TMT10-labeled peptides are isolated by
using an offset (m/z difference between
TMT0 and TMT10).
“
The Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS now supports targeted proteomics assays
with multiplexing in two dimensions: targets and samples. These experiments make use of triggered-byoffset-in-mass peptides that allow for quantitation, even from undetected precursors. Using this method, we
monitored 131 target peptides corresponding to 69 proteins across the NCI-60 cell line in biological triplicate,
analyzing 180 samples in only 48 hours (16 min/sample). We found that accurate and reproducible TMT SPS
tMS3 quantitation elucidated a correlation between expression of key proteins and cellular response to drug
treatment.
—Steven Gygi, Professor, Harvard Medical School
CID MS2
FTMS HCD SPS tMS3
IMPROVED QUANTITATIVE
ACCURACY AT LOW LEVELS
8
Expected
Ratio
Fold Change
6
4
2
m/z
m/z
CID MS2 spectrum of native peptide target
contains multiplexed fragments. Gray bars
indicate the predicted fragment target ions that
are selected for the following SPS tMS3 step.
Targeted SPS MS3 of the predicted
MS/MS fragments generates ten
reporter ions used for quantitation.
0
tMS2
SPS tMS3
Comparison of results for the TMT tMS2 and TMT SPS tMS3
quantification methods shows improved accuracy of the tMS3
method. Each point represents the fold change of a single
quantification event for each of the targeted peptides present
at 100 amol level in matrix. The expected ratio is 8:1.
Advancing Peptide Quantitation
Large Scale Quantitation with Data Independent Acquisition
Protein extraction, and digestion
LC-MS
Creation of MS2
Spectral Library
140 min
HeLa digest standard was introduced into
the Orbitrap Fusion Lumos MS using RP-HPLC.
Data Dependent experiments were used to
generate high resolution accurate mass (HRAM)
MS2 spectra. The data was searched using
SEQUESTTM HT in order to generate a spectral
library for the following DIA data analysis.
3.2 s cycle
Data Independent Acquisition (DIA)
is widely used today for the global
identification and quantification of
thousands of peptides in complex
mixtures. Step-wise isolation and MS/MS
fragmentation of all ions in a defined m/z
window cover the targeted mass range
and provide ultra-high resolution MS and
MS/MS data for all components in the
sample, allowing accurate quantitation
of peptides, and the unique opportunity
for retrospective analysis of unknowns
and new targets of interest in the future.
The Thermo Scientific™ Orbitrap Fusion™
Lumos™ Tribrid™ MS provides speed,
selectivity and enhanced sensitivity
to obtain maximal performance while
maintaining high reproducibility of
quantitation for low abundance analytes.
DATA INDEPENDENT
HRAM HCD MS2
MS scan
m/z 400-1000
Sequential 15 amu-wide
MS/MS scans
400
m/z
1000
DIA acquisition was performed using
a 120,000 FWHM resolution MS scan
followed by 15 amu-wide MS/MS windows
at 30,000 FWHM resolution with 60 ms
maximum injection time.
MATCHING DIA SPECTRUM
TO THE SPECTRAL LIBRARY
QUANTITATION
USING FULL MS
PROTEIN
Protein
Groups
UNIQUE
Unique
Peptides
GROUPS
60,000
PEPTIDES
50,659
50,000
43,143
40,000
30,000
20,000
10,000
6,501
5,964
Spectral
Library
Quantified
by DIA
0
DIA MS2 spectra were searched against
the library built from Sequest HT search
results. Fragment peak detection was
performed with Biognosys SpectronautTM
software.
Peptide quantitation took into account
all confidently identified isotopes from
consecutive MS full scans, correcting
for interference and using cross-run
normalization.
Confident Low-Attomole Limit
Peptide Quantitation Using
Parallel Reaction Monitoring
Parallel Reaction Monitoring (PRM) is uniquely designed
for quantifying hundreds of targeted proteins in complex
matrices. Using this approach, precursor ions are isolated
and selectively fragmented, with the resulting product ions
analyzed in the Orbitrap. This approach benefits from the
brighter ion source and Advanced Quadrupole Technology
of the Thermo Scientific™ Orbitrap Fusion™ Lumos™
Tribrid™ MS, routinely achieving attomole-level limits of
quantification (LOQ) in matrix.
Spectral
Library
Quantified
by DIA
Nearly 92% of the protein groups and over 85%
of the unique peptides were confidently matched
to the spectral library and quantified with <6.9%
CV when using 500 ng of HeLa digest.
GILFVGSGVSGGEEGAR
LTILEELR
GLILVGGYGTR
SAAGAFGPELSR
TASEFDSAIAQDK
LSSEAPALFQFDLK
ELASGLSFPVGFK
NGFILDGFPR
SFANQPLEVVYSK
ELGQSGVDTYLQTK
GISNEGQNASIK
SSAAPPPPPR
DIPVPKPK
IGDYAGIK
HVLTSIGEK
9.8%
10.6%
5.1%
3.3%
100 amol
10 amol
5 amol
1 amol
Fifteen PRTC peptides
were spiked into 200 ng of
HeLa digest and analyzed
by LC-MS (30 min run).
The Orbitrap Fusion Lumos
MS provides accurate
quantitation of all 15 PRTC
peptides with some down
to 1 attomole levels.
Average CV% for each
LOQ level is shown.
“
New Horizons in Intact Protein Analysis
Highly Selective Analysis of Protein Isoforms
“The Lumos proved capable of high selectivity precursor
selection in MS but retained exceptionally high sequence
coverage for MS2. This enabled the decoding of the
most complicated core histone H3 to detect trivalent
proteoforms uniquely detected in a cellular model
of the B-cell cancer, multiple myeloma.”
Top-down mass spectrometry is commonly utilized to characterize intact
proteins and their modifications. The Advanced Vacuum Technology, unique
to the Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS, provides
conditions optimized for improved performance in intact protein analysis.
The high selectivity of Advanced Quadrupole Technology allows for isolation
of precursors and detection of fragments with very high resolving power in
the Orbitrap analyzer. Combined, the new system most efficiently delivers
the highest quality data for the characterization of protein isoforms and their
post-translational modifications.
INTACT HISTONE H3
SPECTRUM, 18+
—Neil Kelleher, Professor, Northwestern University
HIGH RESOLUTION
ISOLATION OF METHYLATED
FORMS OF HISTONE H3
FRAGMENTATION MAPS
OF HISTONE H3 FROM ETD HDTM
TOP-DOWN EXPERIMENTS
14 Da (methylation)
18+
854.9
854.2
853.4
852.6
Dimethylation
Trimethylation
851.8
851.0
850.3
849.4
850
855
m/z
860
865
Advanced Vacuum Technology
increases ion transmission, improving
the quality of intact protein spectra
acquired in the high resolution
accurate mass Orbitrap analyzer.
Zheng, Y et al. Mol Cell Proteomics 2016, 15,776-90.
848
850
Center of 0.5 m/z isolation window
852
854
m/z Dimethylation
856
Trimethylation
With improved ion transmission provided by
the Advanced Quadrupole Technology, it is now
possible to efficiently enrich for individual isobaric
protein forms for a subsequent top-down analysis.
ETD HD identification of isomeric forms of histone H3,
differing in the site of trimethylation (K9 vs. K27) from
the precursors at m/z 853.4.
ETD HD enhances the dynamic range of ETD spectra
by increasing the precursor ion storage capacity.
The higher efficiency of ETD HD experiments provides
greater sequence coverage at faster acquisition rates.
Comprehensive Characterization of Intact Monoclonal Antibodies
Recent advances in protein and antibody-based therapeutics have led to a demand for mass spectrometers
capable of comprehensive characterization of antibody heterogeneity in addition to providing accurate mass
measurements. The Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS allows high accuracy mass
analysis of the intact monoclonal antibody with isotopic resolution of the heavy and light chains. The combination
of the various fragmentation methods available on the Orbitrap Fusion Lumos MS and the improved efficiency
provided by ETD HD enables users to obtain high sequence coverage for the light and heavy chains.
INTACT mAb GLYCOFORMS
7.0
2851.38
24182.85 Da
2830
2860.63
2857.60
2848.28
m/z
2845.40
2803.71
2806.74
2791.81
2754.76
148541.91
2790
2870
931.3
931.5
m/z
931.9
932.1
Glycosylated Heavy Chain, 42+
G1F/G2F
38.6165
148380.82
0.0
G1F/G1F
84.1024
148219.21
3.6
G0F/G1F
100.0000
148058.29
11.8
G0F/G0F
68.152
147909.08
9.2
G0/G0F
10.2611
Average
Mass
TOP-DOWN SEQUENCE COVERAGE
FOR mAb LIGHT AND HEAVY CHAINS
Light Chain, 26+
2854.49
2797.60
2780.64
2794.55
2745.80
2748.82
2751.77
2742.85
2740.03
2750
ISOTOPIC RESOLUTION OF
mAb LIGHT AND HEAVY CHAINS
Matched
Matched
Relative
Delta
Glycoforms Abundance
Mass (ppm)
Raw spectrum showing baseline resolution of
major intact mAb glycoforms. Deconvolution
results are shown in the table above.
49894.17 Da
1189.5
1190.5
50056.48 Da
1191.5
m/z
1192.5
1193.5
Isotopic resolution of light chain (top panel),
and heavy chain (bottom panel), showing the
detection of glycosylated forms.
Combination of ETD HD, CID and HCD fragmentation
modes provided 91% sequence coverage for the light
chain (top panel) and 62% sequence coverage for the
heavy chain (bottom panel).
Revolutionizing Glycoproteomics
Simplified Glycopeptide Analysis
Analysis of post-translational modifications (PTMs) is often central to
biological research because of their roles in cellular function and disease
states. Some PTMs, such as glycosylation, are particularly challenging to
analyze because they require information to both accurately determine the
composition of the modification as well as correctly identify the modification
site. The multiple fragmentation capabilities (CID, HCD, ETD and EThcD) on
the Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS are essential
for high-throughput, comprehensive glycopeptides analysis.
The high dynamic range of ETD and EThcD HD yields high quality MS/MS
spectra of glycopeptide backbone fragmentation, resulting in higher sequence
coverage and accurate localization of the modification site. Accompanying
HCD spectra provide complementary information about glycan composition.
Combination of fragmentation techniques, in addition to intelligent instrument
control workflows for glycopeptide analysis unique to the Orbitrap Fusion
Lumos MS, enables deeper mining of glycosylation modifications in a variety
of biological samples.
EThcD MS/MS spectrum of O-linked glycopeptide
ORBITRAP FUSION LUMOS
CAN PRODUCE:
M+e-NeuAc
HexNAc(1)Hex(1)NeuAc(2)
L A G T E S P V R E E P G E D F PA A R
929.091
M+e-
Large scale LC-MS intact
glycopeptide identification
y13
c13
z10
c2
c3
200
z3
HexNAcHex
HexNAc
c1
z12
y4
400
488.261
y5 z6
600
y6
c15
y15
y7
800
1072.481
z16
y10
m/z
c12
c14
z11
y8
1000
M+2e-NeuAc
y1
z5
Pep 3+
NeuAc-18
NeuAc
c9
c18
c17
M+2e-17
z15
z13
c19
c5
y17
M+2e-Acetyl
y9
z1
Quantitation of intact glycopeptides
using isobaric tags via SPS MS3
M+2e
M+e-17
M+e-Acetyl
c10
Pep+HexNAcHex_2+
Increased peptide sequence coverage
and confidence in glycosylation site
localization with EThcD
HexNAc(1)Hex(1)NeuAc(2)
Pep+HexNAc_3+
Intelligent acquisition strategies
for complete characterization of intact
glycopeptide structure using HCDproduct ion dependent-EThcD/CID
Pep+HexNAcHex_3+
1026.458
c4
y11
1200
c7
1400
1600
c8
1800
Comprehensive Lipid Profiling
Enhancing Productivity with Fast Polarity Switching
For specific lipid sub-classes such as phosphatidylcholine (PC), both positive and negative HCD MS/MS data are required for
full characterization of the individual molecular species. The Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS allows fast
polarity switching and intelligent acquisition of HCD and CID MSn. This provides complementary fragmentation information for
identifying PC molecular species and elucidating co-eluting triacylglycerol (TG) isomers within a single LC-MS run, yielding higher
2
2
ddMS
OT HCD
2
ddMS
OT
15,000
2
throughput, increased sensitivity and more confident
analysis
lipid
molecular
species.
2 HCDof
ddMS
OT HCD
15,000
ddMS
OT15,000
HCD 15,000
ddMS
OT HCD
15,000
ddMS2 OT HCD 15,000
ddMS2 OT HCD 15,000
ddMS2 OT HCD 15,000
POSITIVE MODE 0-18 MIN
NEGATIVE MODE 0-18 MIN
HCD MS/MS
of m/z 810.5997
(P_ChO)+
HCD
MS/MS
FA(20:4)-H
FA(20:4)-H
of m/z 854.5917
(P_ChO)+FA(18:0)-H
FA(18:0)-H
FA(18:0)-H
(P_ChO)+
(P_ChO)+
(P_ChO)+
184.0731
184.0731
FA(20:4)-H
303.2329
283.2643 FA(18:0)-H
283.2643
NL[FA(16:2)]+NH3
NL[FA(16:2)]+NH3
NL[FA(16:0)]+NH
3
NL[FA(16:2)]+NH
NL[FA(16:2)]+NH3
3
NL[FA(16:0)]+NH
523.4724
3
519.4407
NL[FA(16:2)]+NH
NL[FA(16:0)]+NH
HCD
MS/MS
NL[FA(14:0)]+NH
NL[FA(18:2)]
792.7076
NL[FA(18:2)]
NL[FA(14:0)]+NH
NL[FA(18:2)] of m/z
NL[FA(16:2)]+NH
NL[FA(16:0)]+NH
NL[FA(16:0)]+NH
519.4407
FA(20:4)-H
303.2329
495.4408
523.4724
523.4724
519.4407
547.4720
NL[FA(14:0)]+NH3
NL[FA(18:2)]
NL[FA(14:0)]+NH
NL[FA(14:0)]+NH3
NL[FA(18:2)]
3
3
547.4720NL[FA(16:2)]+NH
495.4408
NL[FA(16:2)]+NH
3
3 3
3
NL[FA(16:0)]+NH
547.4720
495.4408
3
547.4720
495.4408
FA(14:0)-OH 523.4724
523.4724
519.4407
519.4407
211.2055 FA(14:0)-OH
3
3
3
184.0731
283.2643
508.3406
224.0691
224.0691
794.5705
575.5032
685.4446
183.1741 794.5705
794.5705339.2902
183.1741
224.0691
3
m/z
m/z
m/z
508.3406
m/z
794.5705
508.3406
100 150 200 250 300 350
100 400
150 450
200 500
250 300 100
350 150
400 200
450100
250
500 300
400 450
500500 200 600 300 700 400
200 350 300
400 100
800 500
100
200
CID MS/MS
of m/z 810.5997
m/z
523.4724
519.4407
575.5032
339.2902
685.4446
183.1741
685.4446
MG(14:0)-OH
MG(14:0)-OH
285.2429
150
250
100 150 200 100
250
300
350
400 400
450 500
285.2429
(P-Cho)-CH
(P-Cho)-CH
(P-Cho)-CH
250
350
450 800
550 250650 603.5353
100 150 200 250 300 350 400
450 500
200 3-H 300 150
400
500
600
700
3-H300
3-H
200
500450
600
700300 100
800
150
350750 750450
500
100
200
400
500
600
700
800
150
250
300
350
400
500
150
250
350
450
550
650
100 150 200 250 300 350100
400150
450200
500250 300 350100400150450
100
200
300
400
500
600
700
800
100200
200
300
400
500
600
700
800
168.0430
LPC(18:0)-CH
168.0430
168.0430
LPC(18:0)-CH3
LPC(18:0)-CH
3
3
M-CH
M-CH
M-CH
m/z
3
3
3
m/z
m/z
m/z
m/z
375.7514
375.7514
m/z
523.4724
547.4720
495.4408
523.4724
FA(14:0)-OH
519.4407 FA(14:0)-OH
FA(14:0)-OH
211.2055
FA(20:4)-H-CO
FA(20:4)-H-CO2
2
211.2055
FA(20:4)-H
FA(20:4)-HFA(20:4)-H-CO
FA(20:4)-H
211.2055
211.2055
FA(20:4)-H-CO2
FA(20:4)-H-CO
2
2
259.2430
259.2430
303.2329
303.2329
FA(18:2)-OH
259.2430
FA(18:2)-OH NL[FA(18:2)]
NL[FA(14:0)]+NH
NL[FA(14:0)]+NH
NL[FA(14:0)]+NH3
NL[FA(18:2)]
NL[FA(18:2)]
259.2430 303.2329
3
3
259.2430
FA(18:0)-H
FA(18:2)-OH
FA(18:0)-H
FA(18:0)-H
FA(18:2)-OH
FA(18:2)-OH 495.4408 NL[FA(12:0)]+NH
(P_ChO)+
263.2372
3
263.2372
547.4720
547.4720
547.4720
495.4408
495.4408 NL[FA(12:0)]+NH
3
FA(12:0)-OH
M-FA1-FA2+H2O-CH3 FA(12:0)-OH
NL[FA(12:0)]+NH
M-FA1-FA2+H2O-CH
283.2643 M-FA1-FA2+H2O-CH
263.2372 FA(12:0)-OH
283.2643
283.2643
NL[FA(12:0)]+NH3
3
NL[FA(12:0)]+NH
263.2372
263.2372 FA(12:0)-OH 575.5032
184.07313
575.5032
FA(12:0)-OH
M-FA1-FA2+H2O-CH
3
M-FA1-FA2+H2O-CH
3
183.1741 3
FA(14:0)-OH
FA(14:0)-OH
FA(14:0)-OH
3
183.1741
224.0691
575.5032
224.0691
575.5032
575.5032
183.1741
224.0691
183.1741 MG(14:0)-OH
224.0691 183.1741
224.0691
MG(14:0)-OH
211.2055
211.2055
211.2055
FA(20:4)-H-CO2
FA(20:4)-H-CO2
FA(20:4)-H-CO2
MG(14:0)-OH
MG(14:0)-OH
MG(14:0)-OH
285.2429
(P-Cho)-CH3-H
285.2429
(P-Cho)-CH3-H
259.2430
603.5353
(P-Cho)-CH3-H 259.2430
603.5353 285.2429
285.2429
(P-Cho)-CH259.2430
(P-Cho)-CH
3-H
FA(18:2)-OH
FA(18:2)-OH
FA(18:2)-OH
603.5353
168.0430
LPC(18:0)-CH3 285.2429
168.0430 3-H
LPC(18:0)-CH
603.5353
603.5353
3
M-CH3
M-CH3 LPC(18:0)-CH
168.0430
168.0430 3 FA(12:0)-OH
LPC(18:0)-CH
NL[FA(12:0)]+NH
NL[FA(12:0)]+NH
NL[FA(12:0)]+NH3
168.0430
LPC(18:0)-CH
375.7514
3
263.2372
263.2372
263.2372
3
3
M-CH
3
375.7514
508.3406
339.2902
M-CH
FA(12:0)-OH
FA(12:0)-OH
3
508.3406
M-CH
339.2902
M-FA1-FA2+H2O-CH
M-FA1-FA2+H2O-CH
M-FA1-FA2+H2O-CH
3
3 508.3406 3
794.5705
3
3
794.5705
375.7514
339.2902 375.7514
508.3406 375.7514
685.4446
(P_ChO)+
184.0731
283.2643
283.2643
184.0731
184.0731
184.0731
(P_ChO)+
FA(20:4)-H
303.2329FA(18:0)-H
303.2329
303.2329
POSITIVE MODE 18-28 MIN
NL[FA(16:0)]+NH3
NL[FA(16:0)]+NH
3
519.4407
m/z
m/z
m/z
m/z
PC (18:0_20:4)
751.5145
751.5145
751.5145
751.5145
m/z
m/z794.5705
508.3406339.2902
794.5705
600
300
150
700
400
250
800
500
m/z
350
600
339.2902
685.4446
450 800 250
550
700150
650
350
m/z
MG(14:0)-OH
350 285.2429
450
550
550
650
750
250 603.5353
350
450
m/z
750
150450
339.2902
250550
m/z
m/z
m/z
375.7514
685.4446
350650
575.5032
450750
m/z
685.4446
650 603.5353
750
550
650
750
685.4446
550
650
750
CID MS3
of m/z FA(18:2)-OH
NL[FA(14:0)]
NL[FA(14:0)]
FA(18:2)-OH
FA(18:2)-OH
NL[FA(14:0)]
NL[FA(14:0)]
NL[FA(14:0)]
792.7076
>547.4722
NL[FA(14:0)]+NH
NL[FA(14:0)]+NH
NL[FA(14:0)]+NHNL[FA(14:0)] NL[FA(14:0)]+NH
FA(18:2)-OH
NL[FA(14:0)]
NL[FA(14:0)]+NHFA(18:2)-OH
NL[FA(14:0)]
751.5145
FA(18:2)-OH
LPC(20:4)-OH
m/zFA(18:2)-OH
283.2643
m/z 283.2643
M+H
LPC(20:4)-OH
263.2362
263.2362m/z
319.2646
m/z 283.2643
526.3292M+H
319.2646 263.2362
283.2643
m/z
283.2643
M+H
263.2362
319.2646
263.2362
810.6010
319.2646
O18:0 FA, C18H
319.2646
H O18:0 FA,
C526.3292
35 2
273.9171
18 35 2
810.6010
H
O
18:0
FA,
C
810.6010 18 35 2
H 273.9171
O18:0
FA, C
O
O
751.5145810.6010
751.5145 18:0 FA, C18H35O
751.5145 O
O
273.9171
18 35 2 273.9171
273.9171
2
O
O
O
O
LPC(20:4)
O
O
LPC(18:0)
LPC(20:4)
LPC(18:0)
P
LPC(20:4)
LPC(18:0)
P
LPC(20:4) O
O
LPC(20:4)
LPC(18:0)
O
O
P
O
O
544.3391
208.3922
524.3699 LPC(18:0)
544.3391
208.3922
3
P
524.3699
P
3
O
OHO
O
O
O
OH O
N
544.3391
208.3922
ON
O
524.3699
FA(18:2)-OH
O
3
544.3391
208.3922
544.3391
524.3699
3
LPC(20:4)-OH
LPC(20:4)-OH524.3699
LPC(20:4)-OH
H
O
3
OH208.3922 N
547.4722
H
O
OH
N
547.4722
OH
m/zN283.2643
m/z 283.2643
H
O
M+H
M+H m/z 283.2643
M+H
547.4722319.2646
HCO
H
O
263.2362 263.2362
263.2362
547.4722
HCO
H
O
627.5240
547.4722
319.2646
627.5240
526.3292
526.3292
526.3292
CH 319.2646
CH
HCO
HCO
627.5240
HCO
627.5240
O
810.6010
810.6010
810.6010
LPC(18:0)-OH
H
O
18:0
FA,
C
H
O
H
O
18:0
FA,
C
18:0
FA,
C
627.5240
CH
O
LPC(18:0)-OH
CH
CH 18
18 35
2
35 2
18 35 2
273.9171
273.9171
273.9171
O
LPC(18:0)-OH
435.0800
O
LPC(18:0)-OH
435.0800
O
O
O
O
O
O
O
LPC(18:0)-OH
170.5874 435.0800
170.5874
506.3599
506.3599LPC(18:0)
435.0800
170.5874
435.0800
170.5874
LPC(20:4) LPC(18:0)
LPC(20:4)
LPC(20:4)
LPC(18:0)
506.3599
170.5874
506.3599
506.3599
P
P
P
m/z
794.5705
m/zO 794.5705
O
O
O
O
O 208.3922
O
O
NL[FA(14:0)]+NH3208.3922
NL[FA(14:0)]+NH3
NL[FA(14:0)]+NH3
544.3391
544.3391 524.3699
544.3391
208.3922
524.3699
524.3699
m/z
m/zO794.5705
303.2330
m/z
794.5705
m/z 303.2330
m/z 794.5705
N
OH
N
OHN
m/z
303.2330
]
-- 303.2330 O[M-CH
m/z
]OH
[M-CH
283.8380
438.7795
m/z 303.2330
H
O
H
H
O
283.8380
438.7795
547.4722
547.4722
547.4722
3
3
[M-CH
H O]2
20:4 FA, C
283.8380
438.7795
]
[M-CH
H31O2
20:4 FA, C20627.5240
[M-CH
283.8380
438.7795
HCO
20 31 3HCO
283.8380
438.7795
627.5240
627.5240
20:43]FA,HCOC20CHH31O2CH
CH3
20:4 FA,
C20H31OO2H O20:4 FA, C
20 31 2
O
O
LPC(18:0)-OH
LPC(18:0)-OH
LPC(18:0)-OH
150 200
250 300 350 400 435.0800
450 500 550
435.0800
150
200
250
300
350
400
450
500
550
435.0800
250
300
350
400
450
500
550
600
650
700
750
800
170.5874
170.5874
170.5874
250 300 350 400 450 500
550
600
650
700
750
800
506.3599
400 200
450 250
500 300
550 350 400 450 500 550
506.3599500 250
150 200 250 300 350 150
400 200
450 250
500 300
550 350 150
600 350
650506.3599
700 450
750 500
800 550 600 650 700 750 800
400
250 300 350 400 450 500 250
550 300
600 350
650 400
700 450
750 800 550 300
m/z 794.5705
m/z 794.5705
m/z 794.5705
m/z
m/z
m/zm/z
m/z
303.2330
m/z 303.2330
m/z 303.2330
m/z
m/z 20:4m/z
m/z
[M-CH3][M-CH ][M-CH3]m/z
283.8380
438.7795
283.8380
438.7795 m/z
283.8380
438.7795
FA, C20H31O220:4 FA,
C20H31O220:4 FA, C3 20H31O2LPC(20:4)-OH
LPC(20:4)-OH
526.3292
LPC(20:4)-OH
M+H
M+H526.3292
810.6010
526.3292
+
3
2
+
m/z
m/z
m/z
O
O
O
O
O
O
O
H
P
H
O
O O
O
O
O
m/z
526.3292
N
O
OH
N
- 18:0 FA,
C18H36O2
m/z 184.0733
O
m/z
184.0733
+
C5H15NO4P
C5PHO15NO4P+
O
O
OH O
P
O
H
O
+
m/z 506.3605
- 20:4 FA, C20H32O2
O
P
O
O
+
m/z 184.0733O
+
P C5H15NO4P
O
O O
H
O
m/z
506.3605
m/z 506.3605
O2
-O20:4 FA, C20H
m/z
32 184.0733
H32O2 +
- 20:4 FA, C
20 NO
P
C5H
15
4
0 MIN
m/z 526.3292
O
H
O
OH
N
- 18:0 FA, C18H36O2
O
+
N
O
O
+
OH
-
2
3
200
250
m/z 526.3292
m/z 526.3292
m/z 526.3292
- 18:0 FA, C18H36O2
- 18:0 FA, C18H36O2
- 18:0 FA, C18H36O2
m/z 526.3292
m/z 526.3292
- 18:0 FA, C18H36O2
- 18:0 FA, C18H36O2
O
2
O
O
O
H
OH
O
N
+
O
300
OHO
+
OH
N
m/z 506.3605
- 20:4 FA, C20H32O2
Fast polarity switching during LC/MS analysis. A cycle with
one positive and one negative Orbitrap MS scan at 120,000
resolving power takes less than 2 seconds to complete.
O
O
O
O
O
O
H
O
18 MIN
500
300
+
550
350
m/z 319.2648
m/z
547.4721 NH
+
O4
NH4FA, C H O O
14:0
14 28 2
O
O
O
NH4
m/z
547.4721
O
m/z 547.4721
]+ - NH
[M+NH
O
4
]+ -3 NH3 [M+NH
4
14:0 FA,
C
H31NO2
O
H 14
14:0
FA,
C14H31NO2
+
O
m/z 547.4721
[M+NH4]+ - NH3 14:0 FA, C14H31NO2
450 500 550
m/z 319.2648
m/z 319.2648
m/z 547.4721 m/z 547.4721 14:0 FA, C14H28O2
14:0 FA, C14H28O2
m/z 319.2648
O
m/z
547.4721
O +
NH
14:0 FA,4 C14H28O2
O
O
O
O
O
O
400
m/z
m/z
319.2648
m/z 547.4721 14:0 FA, C14H28O2
O
H
O
O
NEGATIVE
450
250
m/z 547.4721 14:0 FA, C14H28O2
14:0 FA, C14H28O2
O
+
400
200
m/z 319.2648
(14:0_14:0_18:2)
m/z
m/z 319.2648
m/z 547.4721 -
+
H
O
m/z
506.3605
m/z 506.3605
m/z 506.3605
H32O
O- 20:4 FA, C20
184.0733
184.0733
2
H
O
- 20:4 m/z
FA, C
- m/z
20:4
FA,
C + H32O2
20 32 2
C5H15NO4P+
C5H15NO4P20
m/z 506.3605
- 20:4 FA, C20H32O2
350150400200450250500300550350
150
O
P
m/z 526.3292
ON
O
OH
FA, C18H36NO2
H
O - 18:0
m/z 184.0733
m/z 184.0733
C5H15ONO4P+
C5H15NO4P+
P
O
O
O
3
TG
m/z
O
P
O
O
O
O
3
+
-
3
150
250 300 350 400 450 500
250550
300600
350650
400700
450750
250
500800
300
550 350
600 400
650 450
700 500
750 550
800 600 650 700 750 800
2
+
-
PC (18:0_20:4)
-
3
3
3
2
+
-
2
-
-
2
+
+
+
-
2
O
H
O
O
m/z 319.2648
NH
m/z
547.4721
4
O +
O
NH
14:0 FA, C14H4 28O2
O
H
O
H
O +
O
NH O Om/z 547.4721
NH4
O
m/z 547.47214
m/z 547.4721
]+ - NH3 - +
[M+NH
4
O
[M+NH4]+ - NH3 [M+NH4] - NH3 14:0 FA,
C H31NO2
O14:0
O
H
H 14
FA, C14H31NO2
14:0 FA, C14H31NO2
O
O
+
O
+
m/z 547.4721
[M+NH4]+ - NH3 14:0 FA, C14H31NO2
O
m/z 547.4721
[M+NH4]+ - NH3 14:0 FA, C14H31NO2
POSITIVE
28 MIN
Small Molecule Analysis
Ultimate Confidence in Identification and Quantitation
The high sensitivity and high resolution of the Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ MS makes it a powerful instrument for
the identification and quantitation of small molecules. Due to the instrument’s very high resolution capabilities, fine isotopic structure can
be observed, enabling the determination of highly accurate molecular formulae. This direct measurement removes the ambiguity of pattern
matching estimations and is critical in cases where monoisotopic elements like fluorine or phosphorous may be present in the compound, as
shown in the example of norfloxacin below. Furthermore, the enhanced sensitivity of the Orbitrap Fusion Lumos MS enables far lower levels of
quantitation.
Improved Low Limit
Of Quantitation
NH 4
OH
HO
O
S
Relative
Abundance
Relative
Abundance
Relative
Abundance
O
320.1398
O
Resolving Isobaric
Interferences
HN
N
N
O
F
O
O
100
OH
320.1398
60
20
321.1432
100
322.1462
60
20
320.0
320.5
321.0
100
321.5
320.0
60
321.1432
322.0
320.5
322.1462
321.0
322.0
20
2.6
2.8
3.0
3.2
3.4
13 C13 C
3.6
500 fg
100 fg
Standard
Ion Source
New Ion Source on
Orbitrap Fusion Lumos MS
322.1462
322.1462
18 O18 O
322.1440
322.1440
15 N15
13N
C13 C
15 N15 N
322.1400
322.1400
322.1373
322.1373
Extracted ion chromatogram of Irganox 1035 (M+NH4)+ ion at m/z 660.429 (100 fg on
column). Irganox is a plasticizer known to leach into foods stored in plastic and must be
quantified at very low levels. The Orbitrap Fusion Lumos MS operated in SIM mode was
able to quantify Irganox 1035 in food simulant matrix, achieving an LOQ of 100 fg with
linear dynamic range of 5 orders and <10% CV for all levels. This LOQ is 5x lower than
previously achieved on an Orbitrap instrument equipped with a standard ion source.
2 X213XC13 C
321.1432
321.1432
Time (min)
LOQ
321.5
321.14
321.14
321.15
321.15
m/z
m/z
322.14
322.14
2H 13
2H
C13 C
322.1492
322.1492
322.15
322.15
m/z
m/z
High resolution MS2 spectra of norfloxacin. The direct observations of fine isotopes
of the drug are essential for determining elemental composition.
The list of the
viewable parameters
in the scan and filter
properties can now
be customized
MS OT
TMT SPS MS3
Run advanced
experiments
effortlessly
with validated
template methods
for a variety of
applications
ddMS3 OT HCD
Edit parameters in
easily accessible
properties
panels, featuring
recommended
defaults for each
experiment
Focus on Your Science, Not on Instrument Set Up
The highly intuitive method editor features a user-friendly interface that includes
optimized pre-designed templates for a wide range of applications.
Building on
Revolutionary
Tribrid Architecture
The Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ mass spectrometer further
amplifies the power and versatility of the innovative Tribrid design, first pioneered on
the Orbitrap Fusion MS. The Orbitrap Fusion Lumos mass spectrometer incorporates
the latest technologies and groundbreaking innovations in ion transmission,
dissociation and detection. The combination of these improvements makes it the
most sensitive, most selective and most versatile mass spectrometer to date.
With the Orbitrap Fusion Lumos MS, we are
true partners, committed to innovations to
advance your scientific pursuits.
ADVANCED ACTIVE
ION BEAM GUIDE
Prevents neutrals and high
velocity clusters from entering
mass resolving quadrupole
EASY ETD SOURCE
Based on Townsend discharge;
reliable and easy to use
ELECTRODYNAMIC
ION FUNNEL
Focuses ions after High
Capacity Transfer Tube;
broad tuning curves
DUAL-PRESSURE
LINEAR ION TRAP
n
MS and sensitive mass analysis
of fragments resulting from CID,
ETD HD
HCD, ETD and EThcD
Improved dynamic range
and detection limits for
ETD/EThcD events
ION ROUTING MULTIPOLE
ULTRA-HIGH FIELD
ORBITRAP ANALYZER
Enables parallel analysis; allows
n
HCD at any MS stage
Offers resolution >500,000 FWHM and
scan rates up to 20 Hz at 15,000 FWHM
ADVANCED VACUUM TECHNOLOGY
Reduces pressure in UHV region, improving
transmission to the Orbitrap analyzer
ADVANCED QUADRUPOLE TECHNOLOGY
Segmented design improves transmission at higher resolution;
symmetric transmission across the isolation window
HIGH CAPACITY TRANSFER TUBE
Increases ion flux into the mass spectrometer
Pushing New Frontiers in Science
The Industry’s Leading Portfolio of Mass Spectrometry Solutions
Metabolomics
Non-targeted
Analysis
Exactive Series MS
Tribrid Series MS
Proteomics
Proteomics
Metabolism
HRAM
Bioanalysis
Metabolomics
Lipidomics
Transform
Your Science
Triple Quadrupole MS
Qualitative
Ion Trap MS
PTM Analysis
MS, MS
n
Environmental
Research
Markets
Applied
Markets
Lipidomics
Quantitative
Food Safety
Biomarker Discovery
Lipidomics
Clinical Research/
Forensic Toxicology
Proteomics
Targeted
Analysis
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES
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