non-target analysis with high-resolution mass spectrometry
TRANSCRIPT
Atsushi Yamamoto, Tottori University of Environmental Studies, Japan
Non-target analysis with high-resolution mass spectrometry for environmental samples
1Seminar and Workshop: Environmental Protection, Prevention, Monitoring and RemediationBelgrade University 19 Nov 2018
Tottori University of Environmental Studies
2
Tokyo 13,513,734Kanagawa 9,127,323Osaka 8,838,908
Tottori 573,648Shimane 694,188
Population
Japan
Tottori
••••
•
47 prefecture’s in Japan
47th
Prawn
Oyster
Crab
Starting-up of laboratory
3Now the laboratory is ready for environmental analysis!
Agilent 1260 Infinity/SCIEX API2000
2 liquid chromatograph/mass spectrometersSCIEX EXION LC/X500R QTOF
3 gas chromatograph/mass spectrometersAgilent 7890/5977Thermo Trace GC Ultra/PolarisQHP 6890/5973 controlled by
Windows NT!
State of the art
The first mass spectrometry (MS)
4
• J.J. Thomson achieved separation of ions of different mass within a beam.
• He confirmed that neon could exist in two forms.
What is MS?
5
“ Mass spectrometer separates ions within a beam according to m/z values and measures their relative abundances.
“The dimensionless quantity formed by dividing the ratio of the mass of an ion to the unified atomic mass unit, by its charge number (regardless of sign).Italicized lowercase letters with no spaces.
IUPAC Recommendation 2013
m/z
Mass spectrometer component
7
“ Mass spectrometer separates ions within a beam according to m/z values and measures their relative abundances.
“Ionization source Mass filter Detector
Ionization of substances Selection of an ion of a specified m/z value Detection as current
Measurement of m/z
8
“ Mass spectrometer separates ions within a beam according to m/z values and measures their relative abundances.
“m/z
Ions observed show m/z distribution.
Ability of a measurement to distinguish two peaks.
m/z
100 101 Mass resolution 100
Definition of resolution in MS
9
Ability of a measurement to distinguish two peaks: Resolution
The IUPAC definition of resolution in mass spectrometry expresses this value as m/Δm, where m is the mass of the ion of interest. So, how do we define Δm?
• Peak width definition
(50% → FWHM)
m/z
Δm 50%
50%
Full width at half maximum
• Spacing between two equal intensity peaks with a valley between them(x% Valley)
m/z
Δm 10%
10%
The resolution by FWHM is 2-fold higher than that by 10 % valley.
Example of resolution
10
• Separation of Br – isotopes79 81
m/z
10%
for m/z 79 Resolution10% Valley 39.5
FWHM 83.6
• Separation of [M – H]– generated from hexabromocyclododecane (C12H18Br6) in negative-ion ESI.
640.637509
625m/z
660
10000080
10000001000
[C12H1779Br581Br]–× 500
[C1013CH162H79Br6]–
[C1013C2H1779Br6]–
Ultimate resolution discovers numerous types of matter which we could normally not find. But we want to know optimal resolution for each purpose.
Highresolution (HR)
Performance of MS
11
555 SRM transitions in 1 sec!
Mass resolution10 000 000!
VerapamilS/N 15.8 at 100 ag!
MS satisfies requirements in trace analysis. MS becomes the essential technique in environmental analyses.
• Sensitivity, dynamic range
• High speed analysis
• Mass range
• Mass resolution
Distinguishable as never before
Type of mass analyzers
12
Mass range Speed
Linear dynamic
rangeSensitivity Mass
resolution Founder
Magnetic sector ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ Thomson
1911
Quadrupole ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ Paul1953
Ion trap ✓ ✓ ✓ ✓ ✓ Paul1953
Time of flight ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ Stephen1946
Fourier transform-
ion cyclotron resonance
✓ ✓ ✓ ✓ ✓ ✓ Marshall1974
HR
HR
HR
Terminology
13
Definition in MS Important in HR MS
a Isobar Same nominal mass
b Isotopomer Same numbers of each isotopic atom
c Isotopologue Differ only in the isotopic composition ✓
d Principal ion Most abundant ion of an isotope cluster
e Monoisotopic ionExact mass of an ion calculated using the mass of the most abundant isotope
✓
f Isotopic ion Other ions in an isotope cluster
g Isotope patternSet of peaks related to ions with the same chemical formula
✓
Mass defect (chemistry)
Difference between the nominal mass and the monoisotopic mass
✓
Exact mass Calculated ✓Accurate mass Experimentally determined ✓
d
c
a
b
CO2 ↔ N2OC2H2=CH2
CH2H=CH2H
↔
C12H17Br6
e
f
g
What are we going to do using HR
14
Target analysis
• Specific analyte. Ultra-trace quantitative analysis by definitive separation using HR.
• e.g. dioxin analysis.
Comprehensive analysis
• Prior information such as exact mass values of numerous substances and precursor structures.
Comprehensive analysis • Reading of molecular structure by interpretation
of accurate mass.
Suspect screening
Non-target screening
Analysis X500R QTOF
15
• Hybrid tandem mass spectrometer consists of quadrupole and time of flight.
• Mass resolution and mass accuracy are 30k and 2ppm, respectively.
Quadrupole (Q) type mass spec
16
• Pairs of hyperbolic (or cylindrical) electrodes with DC and AC voltage settings.
–φ0
+φ0+φ0
–φ0x
y z
Attractive and repulsive forces are alternately subjected to ions in x and y directions.
Quadrupole electrode
Quadrupole electrode
Ions with specific mass that stay in stable orbit under certain direct and RF voltages can reach the detector.
✓ Instrument can be compact.✓ High quantitative performance.✓ Rapid scanning of voltage.
Ion
sour
ce
Dete
ctor
Time of flight (TOF) type mass spec
17
Ion source
Acceleration voltage
Flight pathL
V
Mass of ion! : mVelocity of ion! : vCharge number! : zElementary charge! : e
12mv2 = zeV
v = 2zeVm
T =Lv= L m
2zeV
Time of Flight (TOF)
✓ Stable abundance in high mass region.✓ No limitation of mass range.✓ Short measuring time.
Dete
ctor
Tandem mass spectrometry
18
Magnetic sector
Quadrupole
Time of flight
Magnetic sector
Quadrupole
Ion trap
Time of flight
FT-ICR
Ion trap
FT-ICR
Tandem in space
Tandem in time
• Combination of mass analyzer enables consecutive separation of an ion.
When we dissociate an ion, we can know mass values of the fragment (product ion).
Collision cell between mass filters
19
Neutral gas (Nitrogen)
Transit time is over 10–5 sec.
:
H3CC
CH3
O
H3CC
CH3
O
H3CC
CH3
O
Energy needed in dissociation of covalent bond is150-500 kJ/mol → 1.6-5.2 eV
Q is used as ion guide.All ions can pass through Q
Ions gradually increase their internal energy by multiple collisions.
Internal energy repeat convergence and dispersion inside molecule.
The bond with the lowest bond energy dissociate first.
8 IL LI CF I C F1 C C F CF LC C C F
Advantage of QTOF combination
20
• HR product ion spectrum by TOF can be acquired after separation of a precursor ion by Q and following fragmentation in collision cell.
• Rapid acquisition of TOF enables two different acquisition modes. Considering time scale of chromatography, plural acquisition can be regarded as consecutive. (Data dependent acquisition, DDA or IDA; SWATH)
Magnetic sector
Quadrupole
Time of flight
Magnetic sector
Quadrupole
Ion trap
Time of flight
FT-ICR
Ion trap
FT-ICR
MRMHR mode for trace quantitative analysis
21
Pairs of precursor and fragment ions are known for target analytes. MRMHR utilizes this prior information. TOF acquires mass spectrum around mass value of fragment with a range of ± 10 Da.
Quantitation is carried out with extracted ion chromatogram using exact mass of targeted fragment ions.
←20 Da→ ←20 Da→
Around mass values of product ions
←20 Da→ ←20 Da→
←20 Da→ ←20 Da→ Alternating voltage
Dire
ct v
olta
ge
IDA mode for unknown identification
22
Information dependent acquisition
m/z
Once Q forces all ions past, TOF analyzed all ions.
TOF(MS) 2 5 1 3 4
TOF
Q1
For ions with enough abundance, Q reselect the ions. After fragmentation in collision cell, TOF acquires fragment ion spectrum for the reselected precursor ion.
IDA mode for unknown identification
22
Information dependent acquisition
TOF(MS/MS)1~5
m/zm/z
Q 2 5 1 3 4
TOF(MS) 2 5 1 3 4
CIDQ1
TOF
1
For ions with enough abundance, Q reselect the ions. After fragmentation in collision cell, TOF acquires fragment ion spectrum for the reselected precursor ion.
IDA mode for unknown identification
22
Information dependent acquisition
TOF(MS/MS)1~5
m/zm/z
Q 2 5 1 3 4
TOF(MS) 2 5 1 3 4
CIDQ1
TOFIons without enough abundance
will be ignored.so...
1
SWATH
23
Sequential window acquisition of all theoretical fragment-ion spectra
Instead of a single precursor ion, Q selects ions with a mass window (width). TOF acquires spectrum of fragment ions generated from ions that pass Q. Changes of the mass window by Q can enable acquisition of all transition reactions of precursors.
1 2 3 4 5 6 7 8 9 10 11 12
m/z
Q TOF(MS/MS)1~12
m/z
CID
Window
Q1
TOF
SWATH setting in non-target analysis
24
Window 1 Window 2 Window 3 Window 4
Window 1 Window 2 Window 3 Window 4 Any of windows include all Isotopologs.
Isotope information of fragments is available.
MS/MS spectrum loses isotope information.
Isotopologs sometimes divide into two windows.
Window overlap
Isotope pattern
25
0 1 2 3 4 5 6 7 8 9 100 1 2 3 4 5 6 7 8 9 100 1 2 3 4 5 6 7 8 9 10
The number of 13C
C20
C30
C40
C50
C60
C70
C80
C90
C100
• The ratio of A + 1 to A is approximate to the number of carbon atoms included in the molecule.
• Change of isotope pattern during fragmentation.
[A] ion [A+1] ion(monoisotopic ion)
0 1 2 3 4 5 6 7 8 9 100 1 2 3 4 5 6 7 8 9 100 1 2 3 4 5 6 7 8 9 10
Isotope pattern
26
[A] ion [A+2] ion(monoisotopic ion)
Cl2
Cl3
Cl4
Cl5
Cl6
Cl7
Cl8
Cl9
Cl10
• S, Cl, Br have characteristic isotope pattern.
The number of 37Cl
What accurate mass uncover
27
Mass values of atoms that compose molecules is not integer value. Elemental composition can be obtained from accurate mass of molecules.
ElementElement MassMass Majorisotope12C Carbon 12.0000 13C(1.1%)1H Hydrogen 1.0078
16O Oxygen 15.994914N Nitrogen 14.003119F Fluorine 18.998431P Phosphorus 30.973832S Sulfur 31.9721 34S(4.3%)35Cl Chlorine 34.9689 37Cl(24.2%)79Br Bromine 78.9183 81Br(49.3%)
e electron 0.00055
Index for determination of formula
28
• Summation of number of cyclic structures, double bonds, and triple bonds.• aka IHD, index of hydrogen deficiency.
RDB = (4 valence atom) – 1/2 × (1 valence atom) + 1/2 × (3 valence atom) + 1
RDB = (carbon) – 1/2 × (hydrogen and halogen) + 1/2 × (nitrogen) + 1
NH2
OH
O
RDB = 8 – 8/2 + 1 = 5 RDB = 9 – 11/2 + 1/2 + 1 = 5
In cases of LC/ESI-MS, ions generally have fractional number of 0.5. Neutral molecules and neutral losses are integers.
RDB, ring and double bond equivalent
PhenylalanineMetatoluic acid
Trace analysis in environment
29
Although the approach can quantitate extreme trace amount, overall risk assessment is not alway possible.
Selective analysis
Targets are definitive. Cumbersome concentration and cleanup processes.
An enormous number of substances
30
144 000 000
21 000 Substances we generate to a certain amount.Substance registrations (ECHA)
462 Substances enumerated for emission and transfer.Law concerning Pollutant Release and Transfer Register (Jpn)
27 Substances monitored constantly (Aquatic envi)Water quality standard for protection of health (Jpn)
Substances we know.Organic and inorganic substances registrations (ACS)
Non-regulated substances
31
• Not considered a cause of pollution so far, or newly used.
• Become detectable by advance of analytical methods and instruments.
• With proven impact on wildlife.
PharmaceuticalsNano-materials
Fluoroorganicsetc...
Siloxane
There are numerous substances besides regulated substances.
Neat liquid of shampoo (17 000 L) spilled into sewage system from a chemical factory. (2015 Mar)Effluent from the sewage treatment plant intensely foamed and flowed in water resource river.
The local government tested the river water quality and confirmed no detection of regulated substances.
Effluent from the identical plant foamed again. (2015 May)No detection of regulated substances was confirmed.
Accidental water pollution
32
Non target analysis of risk substances
33
Is it possible to comprehensively analyze chemicals to prevent potential risk from unidentified risk substances?
HR-MS is the most promising technique to find unidentified risk substances.
Analytical condition
Difference in mass spectrometer
Isomer
Lack of mass spectra library
Mass spectrum Substance
non one-to-one correspondence
Our society is overflowing with chemical substances. They might include risk substances.
Sampling and pretreatment
34No golden standard for non-target analysis. Ideally, all substances must be grabbed and analyzed.
Ions detected from an atmospheric sample
35
PM2.5 MeOH extract obtained by high volume air sampler with impactor.
m/z
50
800
600
400
200
Retention time / min0 10 20 30 55
Column
Mobile phase
Flow rate
Gradient
GL Science Inertsil ODS-3GL Science Inertsil ODS-3GL Science Inertsil ODS-3A: H2OB: MeOHA: H2OB: MeOHA: H2OB: MeOH0.3 mL/min Injection vol 10 μm0 min (B5%), 30 min (B95%), 35 min (B100%), 45 min (B100%), 45.5 min (B5%), 55 min (B5%)0 min (B5%), 30 min (B95%), 35 min (B100%), 45 min (B100%), 45.5 min (B5%), 55 min (B5%)0 min (B5%), 30 min (B95%), 35 min (B100%), 45 min (B100%), 45.5 min (B5%), 55 min (B5%)
ESI(–)
Ions detected in an aquatic sample
36
• Sample extracted from river water by solid phase extraction cartridge.
m/z
100
1500
1000
500
Column
Mobile phase
Flow rate
Gradient
TOSOH TSK-GEL ODS-100STOSOH TSK-GEL ODS-100STOSOH TSK-GEL ODS-100SA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solutionA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solutionA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solution0.2 mL/min Inj volume 10 μm0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)
ESI(+)
10 20 23Retention time / min
0
m/z 515.4124
Data processing by SCIEX OS
37
515.4124 516.4165 517.4158
ObservedTheoretical
Molecular formula
candidates
Formula Finder
Molecular formula derived from accurate mass and isotope pattern are presented.
Data processing by SCIEX OS
38
Structural information is retrieved from
external database.
Formula Finder
Data processing by SCIEX OS
39
143.0161
Partial structure of C6H7O2S+ is shown in bold style
Bold expression of fragment structure can be used in confirmation of identification.
Fragment Pane
C12H25O S
O
OC12H25
O
Tools for mass spectra interpretation
40
MAGMa
MetFrag/MetFusionISIS
SIRIUS MSMSF
ilter
FingerID
Partitioning
CFM-ID
Simulation of fragmentation using molecular structure database and comparison observed MS/MS spectrum...
Ions detected in an aquatic sample
41
• Sample extracted from river water by solid phase extraction cartridge.
Column
Mobile phase
Flow rate
Gradient
TOSOH TSK-GEL ODS-100STOSOH TSK-GEL ODS-100STOSOH TSK-GEL ODS-100SA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solutionA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solutionA: 5 mM ammonium acetate aqueous solutionB: 5 mM ammonium acetate methanolic solution0.2 mL/min Inj volume 10 μm0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)0 min (B10%), 3 min (B10%), 8 min (B100%), 18 min (B100%), 18.1 min (B10%), 23 min (B10%)
ESI(+)
10 20 22Retention time / min
2
m/z 369.1232 m/z
150
500
300
200
400
Examination of fragment ions
42
• Product ion spectra were converted to a list of m/z values and their intensity.
m/z 369.1232
369.1232
327.0759 369.1232
91.0596243.0576
261.0684
Tools for mass spectra interpretation
43
‣ 8 C F‣ /F C F C C F‣ I C C F IC IC
B . C IA A
3F L LIF CF F C
Tools for mass spectra interpretation
44
‣ 8 C F‣ /F C F C C F‣ I C C F IC IC
B . C C B 5 2I A0
3F L LIF CF F C
Output from the tool
45
/ CAF I A F B MF MC B II M
8C L IL
OP
O
O
O
II
Verification of retrieved result
46
I II
III IV
OP
O
O
O
OP
O
O
O
OP
O
O
O
OP
O
O
O
m/z 369.1232m/z 327.0786
m/z 411.1725m/z 453.2195
The detection of congener series in accurate mass enhances reliability of the retrieved result.
I
IV III
II
Retrieval by the tools
47
• Vast retrieval using big database is possible.• Database can be selected from PubChem,
ChemSpider, KEGG, and HMDB...• The candidates are not always united.• Development of tools is still continued but
automation of MS/MS interpretation will take more time.
Present library or database
48
- 7 71
(
-.58 58 4C I I962 6I C I
O , C C F -) C C F
Analytical example of leachate
49
• Leachate from a waste repository
m/z
200
3000
2000
1000
Retention time / min0 10 20
Column
Mobile phase
Flow rate
Gradient
TOSOH TSK-GEL ODS-100S 5 μm, 2.0 × 150 mmTOSOH TSK-GEL ODS-100S 5 μm, 2.0 × 150 mmTOSOH TSK-GEL ODS-100S 5 μm, 2.0 × 150 mmA: 2 mM Ammonium bicarbonate aq solnB: 2 mM Ammonium bicarbonate methanolic solnA: 2 mM Ammonium bicarbonate aq solnB: 2 mM Ammonium bicarbonate methanolic solnA: 2 mM Ammonium bicarbonate aq solnB: 2 mM Ammonium bicarbonate methanolic soln0.2 mL/min Inj volume 10 μL
0 min (B85%), 10 min (B100%), 16 min (B100%), 16.1 min (B85%), 20 min (B85%)0 min (B85%), 10 min (B100%), 16 min (B100%), 16.1 min (B85%), 20 min (B85%)0 min (B85%), 10 min (B100%), 16 min (B100%), 16.1 min (B85%), 20 min (B85%)
ESI-Negative
Difference of detected ions
50
• Plural adducts might be generated.• Adduct generation usually depend on instrument and condition of ion source.• Discrimination between an ion with additional H and an ion with one more 13C is
difficult by a resolution of 30k.
1574.2857
1637.2872
1650.2716 1574.2814
1636.2848
1650.2943
X500R HR-MS from another vendor
218.9874 311.9760
1217.2753860.2722
Analytical example of leachate
51
• Three fragments with a constant interval of m/z 357.
• m/z 219 and 312 were found in product ion spectrum.
SWATH data includes isotope pattern.
Product ion spectrum of 1574
SWATH
1574.2857
Monoisotopic mass A A+1 A+2 A+31574.2809 100 54 26 9.01217.2753 100 46 18 3.5311.9734 100 5.0 3.9218.9850 100 4.1 0.1)
218.9874 311.9760
1217.2753860.2722
Analytical example of leachate
51
• Three fragments with a constant interval of m/z 357.
• m/z 219 and 312 were found in product ion spectrum.
SWATH data includes isotope pattern.
Product ion spectrum of 1574
SWATH
Product ion spectrum of
1574
1574.2857
Monoisotopic mass A A+1 A+2 A+31574.2809 100 54 26 9.01217.2753 100 46 18 3.5311.9734 100 5.0 3.9218.9850 100 4.1 0.1)
218.9874 311.9760
1217.2753860.2722
Analytical example of leachate
51
• Three fragments with a constant interval of m/z 357.
• m/z 219 and 312 were found in product ion spectrum.
SWATH data includes isotope pattern.
• m/z 219 and 312 have large negative mass defect. The difference of isotope pattern suggest a loss of characteristic element in A+2 such as sulfur.
Product ion spectrum of 1574
SWATH
SWATH
1574.2857
Monoisotopic mass A A+1 A+2 A+31574.2809 100 54 26 9.01217.2753 100 46 18 3.5311.9734 100 5.0 3.9218.9850 100 4.1 0.1
C4F9–92.9886 CH3NSO2
)
Analytical example of leachate
52
218.9850311.9734
1574.2809
– 357.0074 – 357.0076
– 357.0056
Accuratemass Formula Exact
mass
219 218.9850 C4F9– 218.9862
312 311.9734 C5H3NO2F9S– 311.9746
357 C7H8NO3F9S 357.0081
C4F9S
-N
O
O
N-Methyl nonafluorosulfonamide
C4F9S
N
O
O
OH
N-Methyl nonafluoroslfonamido ethanol (N-MeFBSE)
(N-MeFBSA)
Presence of a structure with fluorinated butyl chain
Current regulation of PFASs
53
• Voluntary regulation of F industry. Switch to replacement with short alkyl chain.• It seems to be still used in several countries.
2009.5 Stockholm Convention on POPs (persistent organic pollutants)
2010.42011.4
Revision of Act on the Evaluation and Regulation on Chemical SubstancesPFOS Class I Specified Chemical SubstancesPFOA Chemical Substances Subject to Type II Monitoring
F C SF
FO
OH
O
n
F C CF
F
n
OH
O
F CF
FC C C C C C CF
F
F
F
F
F
F
F
F
F
F
FS
F
F OO
OHF C
F
FC C C C C C CF
F
F
F
F
F
F
F
F
F
F
F OH
O
Perfluoroalkane sulfonic acid (PFAS) Perfluoroalkyl acid (PFAA)
n-PFOS n-PFOA
Production method of F-substances
54
Electrochemical fluorization
SulfonylfluorideCarnonylfluoride
H3C
H2C
CH2
H2C
CH2
H2C
CH2
H2C
SO2F
H3C
H2C
CH2
H2C
CH2
H2C
CH2
COF
Production method of F-substances
54
Electrochemical fluorization
F3C
F2C
CF2
F2C
CF2
F2C
CF2
F2C
SO2F
F3C
F2C
CF2
F2C
CF2
F2C
CF2
COF
Modified by ethylamine and ethylene carbonate
F3CCF
F2C
CF2
F2C
CF2
F2C
SO2F
CF3
F3CCF2
F2C
CF2
F2C
CF2
F2C
SO2F
C8F17SO2
N
H2CCH3
CH2
CH2OH
C8F17SO2
NH
H2CCH3
F3C
F2C
CF2
F2C
CF2
F2C
CF2
F2C
SO2
N
H2CCH3
CH2
H2C
OP
OH
O
OCH2
CH2N
CH2H3C
H2C
CH2
O2S
CF2
F2C
CF2
F2C
CF2
F2C
CF2
F3C
One structural example of end articles. Phosphate ester had been used in paper coating.
(N-EtFOSE)
Reading of accurate mass
55
• Subtract three N-MeFBSEs from 1574. Then the remaining mass is 503.2603.
• Many N and O atoms suggest a possibility for amide or urethane.• Assumption of urethane generated from alcohol and isocyanate.• 3 N-MeFBSE means triisocyanate. Given that triisocyanate of isocyanurate and
N-MeFBSE were used...
Formula Exact mass RDB
C22H33N9O5 503.2610 11.0C23H39N2O10 503.2610 5.5
C18H36N10O5P 503.2613 6.5C19H42N3O10P 503.2613 1.0C17H40N6O9P 503.2600 1.5
C38H33N 503.2618 23.0C27H41N2O3P2 503.2600 9.5C21H37N5O9 503.2597 6.0C34H36N2P 503.2622 18.5C21H37N5O9 503.2597 6.0C24H35N6O6 503.2624 10.5C30H39N3P2 503.2625 14.0
Fractional RDB
Carbon number
Structure consistent with 3 N-MeFBSEs
✓✓ ✓✓
✓
✓ ✓
✓
✓ ✓ ✓
Reading of accurate mass
56
N N
NO O
O
C6H12 C6H12
C6H12
NH
HN
HN
O
O
O
O
O O
C2H4
C2H4
C2H4N
N
N
S
S
S
C4F9
O
O
C4F9
OO
C4F9 O
O
C4F9S
N
O
O
OH
+
N-MeFBSE
OCNC6H12
NCO
N N
N
C6H12
OCN
OO
C6H12
NCO O
C6H12
NCOOCN
C6H12NCO
OCNC6H12
NCO
Triisocyanate
R-OH + OCN-R‘ → R-OCONH-R’
Alcohol Isocyanate Urethane
• Synthesis of urethane
[M – H]– Exact mass1574.2867
Accurate mass1574.2809
Detection of degradation product
57
A structure backing the predicted structure was found in the same sample.
F2C S N
O
O
CF2F2CF3C
O
OH
[M – H]– (m/z 369.9801)
m/z 369.9754
m/z 1574.2809
N-MeFBSAA
N-MeFBSE
N N
NO O
O
C6H12 C6H12
C6H12
NH
HN
HN
O
O
O
O
O O
C2H4
C2H4
C2H4N
N
N
S
S
S
C4F9
O
O
C4F9
OO
C4F9 O
O
• In non target analysis, various substances that slip through the current regulations could be found.
• A resolution of 30 k seems to be enough to determine structure of small molecules.
• The presence of consistent congeners or degradation products can be a key in structural determination in environmental samples.
Conclusions
58
Acknowledgement
59
• This study is financially supported by Grant-in- Aid for Scientific Research (A) of Japan Society for the Promotion of Science (JSPS KAKENHI Grant Number 26241026).
• SCIEX‣ Masaaki Ikeda, Ushio Takeda, Chikako Konomi, Yuka Ikoma
Thank you very much for your attention.
8