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Elizabeth Crawford1, Julie Horner2 and Catharina Crone3
6th October 2012
28th Asilomar Conference on Mass Spectrometry
Pacific Grove, CA USA
Food Packaging:Strategies for Rapid Phthalate Screening by
Ambient Ionization MS2
1 IonSense, Inc. Saugus, MA USA2 Thermo Fisher Scientific, San Jose, CA USA3 Thermo Fisher Scientific, Bremen Germany
DART-SVP Ionization Source
2
Key DART Source Parameters
• Source Parameters:• Ionization gasses
• N2 (Standby)
• He (Run Mode)
• Temperature of ionization gas
• Sample introduction speed
• Ions typically observed in DART–MS mass spectra:• [M+H]+, [M–H]–
• M+·, M–· (ionic compounds)
• Use of dopants to promote ionization:
• [M+NH4]+ (ammonia)
• [M+Cl]– (e.g. CH2Cl2)
• [M+CF3COO]– (trifluoroacetic acid)
3
Figure courtesy of Prof. Jana Hajšlová
ICT Prague, Czech Republic
4
Major Regulated Phthalates of InterestCompound Elemental
composition
Precursor
[M+H]+
Selection of characteristic
fragments nominal m/z
Regulation
Di-n-butyl phthalate (DBP) C16H22O4 279.1591 205.1 CA Prop 65/ EU
Diisobutyl phthalate (DiBP) C16H22O4 279.1591 205.1; 223.1, 57.1 EU
Benzyl butyl phthalate
(BBP)
C19H20O4 313.1434 205.1; 239.1; 91.1 CA Prop 65/ EU
Bis(2-ethylhexyl)phthalate
(DEHP)
C24H38O4 391.2843 279.2; 261.1; 167.0; 113.1 CA Prop 65/ EU
Di-n-octyl phthalate
(DnOP)
C24H38O4 391.2843 261.1; 167.0 CA Prop 65/ EU
Dioctyltere phthalate
(DOTP)
C24H38O4 391.2843 262.1; 167.0; 111.1; 280.1 Not Regulated
Diisononyl phthalate
(DiNP)
C26H42O4 419.3156 275.2; 127.1; 293.2 CA Prop 65/ EU
Diisodecyl phthalate
(DiDP)
C28H46O4 447.3469 289.2; 141.2; 307.2; 99.1 CA Prop 65/ EU
Isomeric phthalates are shown in blue and yellow.
5
MS/MS Optimization: Phthalate Screening
Phthalate standards
ordered from Sigma
Aldrich and all (except
DMTP) were directly
sampled as liquids on
closed end glass
capillary tube as
pictured here.
Analyte signal was long
lasting (> 1 minute) and
an MS/MS instrument
method was
constructed to ramp
through activation
energies of 20, 25, 30,
35, 40%.
DMTP standard is a solid and was directly sampled using tweezers.
6
• DART-SVP source angled at a
in line with the API inlet.
• For phthalate screening the
DART source was operated at
a modest 250° C heater temp.
• The sample stage was
automatically moved toward
the DART source at a constant
0.5 mm/s speed and the
sample was positioned
directly in the heated gas
beam.
General Sampling
7
A96-1_A_YellowHandlePiece_20120412_PO... 4/12/2012 10:50:56 AM 3+D; Background BEG&ENDHe; 200C; 1.0 mm/s;1e6 AGC FillRT: 0.00 - 1.57
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5
Time (min)
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative A
bu
nd
an
ce
1.430.56 0.60 1.400.74 1.360.76 1.460.661.320.80 1.50
1.270.84
1.02
0.480.440.360.22 0.290.150.08
1.430.56 0.63 0.73 1.38 1.461.360.76 1.481.320.801.501.27
1.021.00
1.100.440.390.330.290.220.170.08
NL: 7.46E5
m/z= 149.02254-149.02403 MS A96-1_A_YellowHandlePiece_20120412_POS_He_Run71
NL: 2.51E6
m/z= 391.28217-391.28608 MS A96-1_A_YellowHandlePiece_20120412_POS_He_Run71
A96-1_A_YellowHandlePiece_20120412_POS_He_Run71 #47-77 RT: 0.56-0.80 AV: 31 SB: 19 0.05-0.27 NL: 2.04E6T: FTMS {1,1} + p NSI Full ms [100.00-1000.00]
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850
m/z
0
10
20
30
40
50
60
70
80
90
100
Re
lative A
bu
nda
nce
391.28312
798.58545
149.02301
239.14834 279.15841167.03355408.30952 841.59119
Yellow Plastic Handle
DEHP, DnOP and DOP
XIC m/z 391
Major Ortho
Phthalate Fragment
XIC m/z 149
DEHP, DnOP and DOP
Major Thermal
Fragment
8
390.5 391.0 391.5 392.0 392.5 393.0 393.5 394.0
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Rela
tive
Ab
un
da
nce
392.28770
393.29061394.29339
391.28318
392.28644
393.28940
NL:8.93E4
C 24 H 38 O 4 +H: C 24 H 39 O 4
p (gss, s /p:8) Chrg 1R: 50000 Res .Pwr . @FWHM
NL:1.50E6
A96-
DEHP, DnOP, DOP (Isomeric Phthalates)
391.28429[M+H]+ exact mass simulation
DEHP, DnOP and DOP
[M+H]+ ion [C24H38O4 + H]+
δ mass accuracy: -2.8 ppm
9
DART-SVP: Velos Pro Linear Ion Trap
10
Velos Pro - Instrument Method Set-up
11
Mass List: Including Opt. CID Energies
12
EPA Mix 506: Phthalate & Adipate Esters EPA506_PhthalateMix_AllOpt_20121004 #350-409 RT: 1.18-1.33 AV: 6 SB: 7 0.10-0.33 NL: 2.48E7F: ITMS + p NSI Full ms [50.00-500.00]
50 100 150 200 250 300 350 400 450 500
m/z
0
10
20
30
40
50
60
70
80
90
100
Rela
tive
Ab
un
da
nce
149.00
391.26
279.09
223.09
313.09
371.25
177.00
195.0091.08 121.00
DEHP, DnOP
DBP
BBPDEHA
DEP
DMP
13
Optimized MS/MS Method: EPA Mix 506
50 100 150 200 250 300 350 400
m/z
0
50
100
0
50
100
0
50
100
Re
lative A
bu
nd
ance 0
50
100
0
50
100162.98
177.02
148.95
205.01
166.89279.11
205.01
148.9591.06 238.95
261.06
148.95
279.12
113.10 391.28166.96
NL: 2.58E5
EPA506_PhthalateMix_AllOpt_20121004#371 RT: 1.24 AV: 1 SB: 10 0.43-0.78 F: ITMS + c NSI Full ms2 195.00
NL: 2.20E6
EPA506_PhthalateMix_AllOpt_20121004#350-393 RT: 1.19-1.30 AV: 5 SB: 10 0.43-0.78 F: ITMS + c NSI Full ms2 223.00
NL: 7.84E5
EPA506_PhthalateMix_AllOpt_20121004#359-409 RT: 1.21-1.35 AV: 6 SB: 11 0.43-0.78 F: ITMS + c NSI Full ms2 279.00
NL: 4.52E6
EPA506_PhthalateMix_AllOpt_20121004#365-474 RT: 1.23-1.52 AV: 11 SB: 10 0.43-0.78 F: ITMS + c NSI Full ms2 313.10
NL: 1.18E7
EPA506_PhthalateMix_AllOpt_20121004#365-474 RT: 1.23-1.52 AV: 11 SB: 10 0.43-0.78 F: ITMS + c NSI Full ms2 391.10
DEHP/DnOP
Mix
DBP
BBP
DEP
DMP
14
DEHP and DnOP Fragmentation Pathways
DEHP R1 = C4H9
R2 = C2H5
O
O
O
O
R1
R1
R2
R2
H
DnOP R1 = H
R2 = C6H13
DnOP DEHP
m/z 261.1485
O
O
O
OH
O
O
m/z 149.0233
-H2O
-H2O
m/z 279.1591
OH
O
O
OH
m/z 167.0339
OH
OH
O
O
Fragmentation proposed by Rothenbacher T. and Schwack W,
Rapid Commun. Mass Spectrom. 2009, 23:2829-2835
15
Isomeric m/z 391: DOTP, DnOP & DEHP
100 150 200 250 300 350 400
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative A
bu
nd
ance
0
20
40
60
80
100262.14
166.97111.09
280.14
390.36127.08 247.11
261.10
148.98
113.17
148.98 279.14
113.14261.01166.98
NL: 1.68E4
DOTP_Std_CIDOpt_20121004#749-809 RT: 1.89-2.01 AV: 10 SB: 36 0.19-0.73 F: ITMS + c NSI Full ms2 [email protected] [85.00-400.00]
NL: 5.85E7
dnop_std_cidopt_20121004#943-1017 RT: 2.08-2.21 AV: 13 SB: 41 0.19-0.73 F: ITMS + c NSI Full ms2 [email protected] [85.00-400.00]
NL: 1.79E7
dehp_std_cidopt_20121004#823-866 RT: 1.87-1.94 AV: 8 SB: 42 0.19-0.73 F: ITMS + c NSI Full ms2 [email protected] [85.00-400.00]
DOTP
DnOP
DEHP
16
Isomeric Fragmentation: CID vs. HCD
50 100 150 200 250
m/z
0
50
100
0
50
100
0
50
100
Re
lative A
bu
nda
nce 0
50
100204.99
148.97
223.06
205.02
148.96
148.96
279.12
204.99166.9257.09 223.06
148.96
205.01
279.11166.91
NL: 4.88E6
dibp_std_cidopt_20121004#692-770 RT: 1.78-1.90 AV: 13 SB: 20 0.09-0.40 F: ITMS + c NSI Full ms2 [email protected] [60.00-285.00]
NL: 9.88E6
dbp_std_cidopt_20121004#721-939 RT: 1.81-2.14 AV: 36 SB: 20 0.09-0.40 F: ITMS + c NSI Full ms2 [email protected] [60.00-285.00]
NL: 2.34E6
DiBP_Std_HCDOpt_20121004#890-1041 RT: 2.18-2.40 AV: 25 SB: 18 0.09-0.40 F: ITMS + c NSI Full ms2 [email protected] [50.00-285.00]
NL: 2.61E6
dbp_std_hcdopt_20121004#428-636 RT: 1.05-1.35 AV: 34 SB: 19 0.09-0.40 F: ITMS + c NSI Full ms2 [email protected] [50.00-285.00]
DBP Std
DiBP Std
DBP Std
DiBP Std
CID
CID
HCD
HCD
17
HRAM MS/MS HCD Fragmentation
160 180 200 220 240 260 280
m/z
0
20
40
60
80
1000
20
40
60
80
100
Re
lati
ve A
bu
nd
an
ce
205.0856
167.0336 279.1586
167.0336205.0855
223.0960
279.1588
NL: 2.90E6
120831_SIM_HCD_DBP_Spot7a_1#160 RT: 1.41 AV: 1 T: FTMS + p NSI Full ms2 [email protected] [100.00-305.00]
NL: 3.56E5
120831_sim_hcd_dibp_spot5a#128 RT: 1.13 AV: 1 T: FTMS + p NSI Full ms2 [email protected] [100.00-305.00]
Re
lati
ve A
bu
nd
an
ce
O 2H
O
O
O
O
O
O
O
O
O
OH
OH
O
O
H
OH
OH
O
O
H
O
O
O
O
O
O
O
O
DBP Std
DiBP Std
18
Rapid ScreeningFood Packaging, Utensils &
Sports Equipment
19
Olive Jar Lid: Full Scan
50 100 150 200 250 300 350 400 450 500
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Rela
tive
Ab
un
dan
ce
403.26
315.25 361.17 441.34282.25
282.25
338.34
228.17 298.25172.09 354.34256.25 453.34403.17109.08 210.09
NL: 4.40E7
OldOliveLid_AllOpt_20121004_02#433-559 RT: 1.36-1.69 AV: 11 SB: 10 2.56-2.88 F: ITMS + p NSI Full ms [50.00-500.00]
NL: 1.28E7
newolivelid_allopt_20121004#129-206 RT: 0.42-0.61 AV: 7 SB: 9 2.56-2.88 F: ITMS + p NSI Full ms [50.00-500.00]
Older Lid
Newer Lid
Manufacturer
“PVC Free”
tributyl O-
acetylcitrate
(ATBC)
Erucamide
20
Croatian Lid Gasket - Positive DnOP
100 150 200 250 300 350 400
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Rela
tive
Ab
un
dan
ce
261.11
148.98
391.32
261.06
148.94
391.31
373.30
NL: 6.76E7
dnop_std_cidopt_20121004#848-1030 RT: 1.92-2.23 AV: 30 SB: 34 0.88-1.36 F: ITMS + c NSI Full ms2 [email protected] [85.00-400.00]
NL: 5.05E5
CroatianLid_AllOpt_20121004_02#134-234 RT: 0.45-0.68 AV: 10 SB: 5 0.09-0.25 F: ITMS + c NSI Full ms2 391.10
DnOP Standard
Sampled Lid Gasket
21
Las Vegas Styrofoam Cup – Positive Hit
100 150 200 250 300 350 400 450
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative A
bu
nd
ance
0
20
40
60
80
100279.12
148.96
113.10261.03
166.96 391.31
275.06
127.08 293.13
148.9885.18
166.99
307.14
141.06
148.92
166.93
99.08275.09 429.27
NL: 5.80E4
VegasSmoothieCup_AllOpt_20121004#340-385 RT: 1.05-1.14 AV: 4 SB: 3 0.04-0.15 F: ITMS + c NSI Full ms2 391.10
NL: 6.90E6
VegasSmoothieCup_AllOpt_20121004#639 RT: 1.92 AV: 1 SB: 3 0.04-0.15 F: ITMS + c NSI Full ms2 419.10
NL: 1.73E5
VegasSmoothieCup_AllOpt_20121004#644-666 RT: 1.92-1.95 AV: 2 SB: 3 0.04-0.15 F: ITMS + c NSI Full ms2 447.10
DEHP
DiNP
DiDP
22
Coffee Stirrer
50 100 150 200 250 300 350 400
m/z
0
20
40
60
80
100
0
20
40
60
80
100
Re
lative A
bu
nd
ance
0
20
40
60
80
100279.18
148.95
205.0257.12
279.11
148.96
391.32113.09 261.03
166.96373.27
275.05
127.05293.11
148.94418.36
363.1885.12
NL: 6.05E4
AircraftCoffeeStick_AllOpt_20121005#124-156 RT: 0.41-0.46 AV: 3 SB: 8 2.52-2.78 F: ITMS + c NSI Full ms2 [email protected] [50.00-290.00]
NL: 9.71E4
AircraftCoffeeStick_AllOpt_20121005#128-164 RT: 0.43-0.48 AV: 3 SB: 5 0.04-0.21 F: ITMS + c NSI Full ms2 [email protected] [75.00-400.00]
NL: 6.26E4
AircraftCoffeeStick_AllOpt_20121005#124-232 RT: 0.40-0.68 AV: 10 SB: 5 0.06-0.20 F: ITMS + c NSI Full ms2 [email protected] [80.00-425.00]
DiBP
DEHP
DiNP
23
Airline Plastic Fork: DEHP & DnOPAircraftFork_AllOpt_20121005 10/5/2012 8:53:06 AM Aircraft utensil packet250C, He, tweezerRT: 1.25 - 1.93
1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90
Time (min)
0
50
1000
50
1000
50
100 NL: 1.03E6
m/z= 258.79-263.10 F: ITMS + c NSI Full ms2 391.10 MS AircraftFork_AllOpt_20121005
NL: 6.70E5
m/z= 276.58-283.04 F: ITMS + c NSI Full ms2 391.10 MS AircraftFork_AllOpt_20121005
NL: 2.30E5
m/z= 110.57-115.42 F: ITMS + c NSI Full ms2 391.10 MS AircraftFork_AllOpt_20121005
80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400
m/z
0
50
100
0
50
100
Re
lative
Ab
und
ance 0
50
100261.10
148.97
391.31113.11 279.14
261.08
148.98
279.14
113.11 391.35167.01
148.96 279.14
261.05
113.09 391.36166.97
373.37275.18247.15 355.35
NL: 5.87E5
AircraftFork_AllOpt_20121005#505-546 RT: 1.49-1.59 AV: 4 SB: 5 0.04-0.20 F: ITMS + c NSI Full ms2 391.10
NL: 2.99E5
AircraftFork_AllOpt_20121005#556-583 RT: 1.62-1.67 AV: 3 SB: 5 0.04-0.20 F: ITMS + c NSI Full ms2 391.10
NL: 2.69E5
AircraftFork_AllOpt_20121005#589-633 RT: 1.70-1.80 AV: 5 SB: 5 0.04-0.20 F: ITMS + c NSI Full ms2 391.10
DEHP
DnOP
DnOP DEHP
Mix
DnOP
DEHPMore heat into the sample as it stays in heated gas
DEHP
24
Yoga MatYogaMat_AllOpt_20121005 10/5/2012 11:01:09 AM Katarina's yoga mat250C, He, tweezerRT: 0.00 - 1.62
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6
Time (min)
0
50
1000
50
1000
50
1000
50
100 NL: 6.60E7
m/z= 278.50-279.50+390.50-391.50+418.50-419.50+446.50-447.50 F: ITMS + p NSI Full ms [50.00-500.00] MS YogaMat_AllOpt_20121005
NL: 4.63E6
m/z= 260.50-261.50+278.50-279.50 F: ITMS + c NSI Full ms2 391.10 MS YogaMat_AllOpt_20121005
NL: 3.92E5
m/z= 126.50-127.50+274.60-275.60 F: ITMS + c NSI Full ms2 419.10 MS YogaMat_AllOpt_20121005
NL: 4.10E4
m/z= 148.50-149.50 F: ITMS + c NSI Full ms2 447.10 MS YogaMat_AllOpt_20121005
50 100 150 200 250 300 350 400 450 500
m/z
0
50
100
0
50
100
0
50
100
Re
lative A
bu
nda
nce
0
50
100391.26
149.00 279.1761.17
313.17 419.34228.17 379.25335.25195.09177.09 267.17 447.34134.08 466.3486.08
279.11148.96
261.03113.10 166.97 391.33195.11 373.28257.23 285.31 331.3099.12
275.07
127.06 293.13148.95 401.30 419.3985.14 363.17195.09 313.34239.14115.15
307.17289.07429.34141.07
447.41166.96 387.30195.10 341.2599.13 127.06 255.2185.21 239.14
NL: 4.15E6
YogaMat_AllOpt_20121005#170-231 RT: 0.53-0.65 AV: 6 SB: 5 0.08-0.27 F: ITMS + p NSI Full ms [50.00-500.00]
NL: 1.38E6
YogaMat_AllOpt_20121005#170-231 RT: 0.52-0.66 AV: 9 SB: 8 0.06-0.34 F: ITMS + c NSI Full ms2 391.10
NL: 1.55E5
YogaMat_AllOpt_20121005#162-231 RT: 0.52-0.65 AV: 6 SB: 7 0.10-0.33 F: ITMS + c NSI Full ms2 419.10
NL: 3.09E4
YogaMat_AllOpt_20121005#176-236 RT: 0.55-0.65 AV: 5 SB: 7 0.10-0.33 F: ITMS + c NSI Full ms2 447.10
Full Scan
DEHP
DiNP
DiDP
BlackBlue
• Initial screening by DART high resolution MS, need fragmentation
• Developed MS/MS instrument method for confirmation/identification and distinction between isomeric phthalates for rapid screening
• Further work needed in order to investigate migration of plasticizers into various food stuffs for a quantitative approach
• Expand the method to include other additives
• Software program is needed to quickly process and interpret data
25
Summary
Email: [email protected]
![bis 2 ethylhexyl phthalate-eng · Bis(2-ethylhexyl) phthalate [also known as di(2-ethylhexyl) phthalate or DEHP], a branched-chain dioctyl ester of phthalic acid, is the most important](https://img.pdfslide.net/doc/110x75/5ff6bd9cd1b4ca2c50283d75/bis-2-ethylhexyl-phthalate-eng-bis2-ethylhexyl-phthalate-also-known-as-di2-ethylhexyl.jpg)
