non-target analysis with high-resolution mass spectrometry

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

Joey Ramone

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


“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


“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.


22


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.


22


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.


38

Structural information is retrieved from

external database.

Formula Finder


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


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


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


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


51





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


51




• 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.


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

)


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

non-target analysis with high-resolution mass spectrometry

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