Food Packaging: Strategies for Rapid Phthalate ... - IonSense

Food Packaging:
Strategies for Rapid Phthalate Screening by
Ambient Ionization MS 2
Elizabeth Crawford 1 , Julie Horner 2 and Catharina Crone 3
1
IonSense, Inc. Saugus, MA USA
2
Thermo Fisher Scientific, San Jose, CA USA
3
Thermo Fisher Scientific, Bremen Germany
6 th October 2012
28 th Asilomar Conference on Mass Spectrometry
Pacific Grove, CA USA

DART-SVP Ionization Source
2

Key DART Source Parameters
• Source Parameters:
• Ionization gasses
• N 2 (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. CH 2 Cl 2 )
• [M+CF 3 COO] – (trifluoroacetic
acid)
Figure courtesy of Prof. Jana Hajšlová
ICT Prague, Czech Republic
3

Major Regulated Phthalates of Interest
Compound
Elemental
composition
Precursor
[M+H]+
Selection of characteristic
fragments nominal m/z
Regulation
Di-n-butyl phthalate (DBP) C 16 H 22 O 4 279.1591 205.1 CA Prop 65/ EU
Diisobutyl phthalate (DiBP) C 16 H 22 O 4 279.1591 205.1; 223.1, 57.1 EU
Benzyl butyl phthalate
(BBP)
Bis(2-ethylhexyl)phthalate
(DEHP)
Di-n-octyl phthalate
(DnOP)
Dioctyltere phthalate
(DOTP)
Diisononyl phthalate
(DiNP)
Diisodecyl phthalate
(DiDP)
C 19 H 20 O 4 313.1434 205.1; 239.1; 91.1 CA Prop 65/ EU
C 24 H 38 O 4 391.2843 279.2; 261.1; 167.0; 113.1 CA Prop 65/ EU
C 24 H 38 O 4 391.2843 261.1; 167.0 CA Prop 65/ EU
C 24 H 38 O 4 391.2843 262.1; 167.0; 111.1; 280.1 Not Regulated
C 26 H 42 O 4 419.3156 275.2; 127.1; 293.2 CA Prop 65/ EU
C 28 H 46 O 4 447.3469 289.2; 141.2; 307.2; 99.1 CA Prop 65/ EU
Isomeric phthalates are shown in blue and yellow.
4

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.
5

General Sampling
• 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.
6

Yellow Plastic Handle
A96-1_A_YellowHandlePiece_20120412_PO... 4/12/2012 10:50:56 AM 3+D; Background BEG&END
He; 200C; 1.0 mm/s;1e6 AGC Fill
RT: 0.00 - 1.57
80
60
40
20
0
100
Relative Abundance100
80
60
40
0.08
0.15
0.22 0.29
0.36
0.44
0.48
0.56 0.60 1.43
0.74
1.40
0.66 0.76 1.36
1.46
0.80 1.32
1.50
0.84
1.27
1.02
0.56 0.63
1.43
0.73
0.76 1.36 1.38 1.46
1.32
1.48
0.80
1.27
1.50
20
1.10
0.08 0.17 0.22 0.29 0.33 0.39 0.44
0
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)
A96-1_A_YellowHandlePiece_20120412_POS_He_Run71 #47-77 RT: 0.56-0.80 AV: 31 SB: 19 0.05-0.27 NL: 2.04E6
T: FTMS {1,1} + p NSI Full ms [100.00-1000.00]
391.28312
100
Relative Abundance
90
80
70
60
50
40
30
20
Major Ortho
Phthalate Fragment
XIC m/z 149
DEHP, DnOP and DOP
XIC m/z 391
Major Thermal
Fragment
149.02301
1.00
DEHP, DnOP and DOP
1.02
NL: 7.46E5
m/z=
149.02254-
149.02403 MS
A96-
1_A_YellowHandlePi
ece_20120412_POS_
He_Run71
NL: 2.51E6
m/z=
391.28217-
391.28608 MS
A96-
1_A_YellowHandlePi
ece_20120412_POS_
He_Run71
798.58545
10
167.03355 239.14834 279.15841
0
408.30952 841.59119
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850
m/z
7

DEHP, DnOP, DOP (Isomeric Phthalates)
100
391.28429
[M+H] + exact mass simulation
NL:
8.93E4
Relative Abundance
80
60
40
20
392.28770
C 24 H 38 O 4 +H:
C 24 H 39 O 4
p (gss, s /p:8) Chrg 1
R: 50000 Res .Pwr . @FWHM
0
100
80
391.28318
DEHP, DnOP and DOP
[M+H] + ion [C 24 H 38 O 4 + H] +
δ mass accuracy: -2.8 ppm
393.29061
NL:
1.50E6
A96-
394.29339
60
40
20
0
392.28644
393.28940
390.5 391.0 391.5 392.0 392.5 393.0 393.5 394.0
m/z
8

DART-SVP: Velos Pro Linear Ion Trap
9

Velos Pro - Instrument Method Set-up
10

Mass List: Including Opt. CID Energies
11

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: 2.48E7
F: ITMS + p NSI Full ms [50.00-500.00]
149.00
100
90
DEHP, DnOP
80
391.26
70
DBP
Relative Abundance
60
50
40
30
20
10
91.08 121.00
177.00
DMP
195.00
DEP
223.09
279.09
313.09
371.25
0
50 100 150 200 250 300 350 400 450 500
m/z
BBP
DEHA
12

Optimized MS/MS Method: EPA Mix 506
100
50
162.98
DMP
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
0
100
50
177.02
DEP
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
Relative Abundance
0
100
50
0
100
50
0
100
50
0
148.95
166.89
205.01
205.01
148.95
91.06 238.95
113.10
148.95
166.96
261.06
279.11
279.12
DBP
BBP
DEHP/DnOP
Mix
391.28
50 100 150 200 250 300 350 400
m/z
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
13

DEHP and DnOP Fragmentation Pathways
O
O
O
O
R 2
R 2
R 1
R 1
H
DEHP R 1 = C 4 H 9
R 2 = C 2 H 5
DnOP
R 1 = H
R 2 = C 6 H 13
DnOP
DEHP
O
-H 2 O
OH
O
O
m/z 261.1485
O
OH
O
m/z 279.1591
O
O
OH
m/z 149.0233
-H 2 O
OH
O
O
m/z 167.0339
OH
Fragmentation proposed by Rothenbacher T. and Schwack W,
Rapid Commun. Mass Spectrom. 2009, 23:2829-2835
14

Isomeric m/z 391: DOTP, DnOP & DEHP
100
80
60
40
111.09
166.97
262.14
280.14
DOTP
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
391.00@cid25.00 [85.00-400.00]
20
0
80
60
40
20
0
100
Relative Abundance100
80
60
127.08 247.11
261.10
148.98
113.17
148.98 279.14
390.36
DnOP
DEHP
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
391.00@cid25.00 [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
391.00@cid25.00 [85.00-400.00]
40
20
113.14
166.98
261.01
0
100 150 200 250 300 350 400
m/z
15

Isomeric Fragmentation: CID vs. HCD
100
50
DiBP Std
148.97
204.99
CID
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 279.00@cid40.00
[60.00-285.00]
Relative Abundance
0
100
50
0
100
50
DBP Std
DiBP Std
223.06
205.02
148.96
148.96
166.92
204.99
57.09 223.06
CID
HCD
279.12
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 279.00@cid40.00
[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 279.00@hcd10.00
[50.00-285.00]
0
100
50
DBP Std
148.96
166.91
205.01
HCD
279.11
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 279.00@hcd10.00
[50.00-285.00]
0
50 100 150 200 250
m/z
16

HRAM MS/MS HCD Fragmentation
Relative Abundance
Relative Abundance
100
80
60
40
20
100 0
80
60
40
20
0
205.0856
167.0336 O
279.1586
167.0336
O
O
O
OH
OH
OH
OH
H
H
O
O
O
205.0855
O
O
223.0960
279.1588
160 180 200 220 240 260 280
m/z
O
O
O
OH
2
O
DBP Std
O
O
O
O
DiBP Std
O
O
O
O
NL: 2.90E6
120831_SIM_HCD_DBP_S
pot7a_1#160 RT: 1.41 AV:
1 T: FTMS + p NSI Full ms2
279.16@hcd15.00
[100.00-305.00]
NL: 3.56E5
120831_sim_hcd_dibp_spot
5a#128 RT: 1.13 AV: 1 T:
FTMS + p NSI Full ms2
279.16@hcd15.00
[100.00-305.00]
17

Rapid Screening
Food Packaging, Utensils &
Sports Equipment
18

Olive Jar Lid: Full Scan
Relative Abundance
100
80
60
40
20
Older Lid
403.26
tributyl O-
acetylcitrate
(ATBC)
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]
0
100
80
60
40
282.25
282.25
315.25 361.17 441.34
338.34
Erucamide
Newer Lid
Manufacturer
“PVC Free”
NL: 1.28E7
newolivelid_allopt_20121004#1
29-206 RT: 0.42-0.61 AV: 7
SB: 9 2.56-2.88 F: ITMS + p
NSI Full ms [50.00-500.00]
20
0
228.17
172.09
298.25
354.34
109.08 210.09 256.25 403.17 453.34
50 100 150 200 250 300 350 400 450 500
m/z
19

Croatian Lid Gasket - Positive DnOP
Relative Abundance
100
80
60
40
20
DnOP Standard
148.98
261.11
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 391.00@cid20.00
[85.00-400.00]
0
100
80
Sampled Lid Gasket
261.06
391.32
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
60
40
20
0
148.94
373.30
391.31
100 150 200 250 300 350 400
m/z
20

Las Vegas Styrofoam Cup – Positive Hit
100
80
60
148.96
279.12
DEHP
NL: 5.80E4
VegasSmoothieCup_AllOpt_201
21004#340-385 RT: 1.05-1.14
AV: 4 SB: 3 0.04-0.15 F: ITMS
+ c NSI Full ms2 391.10
40
20
113.10
261.03
166.96 391.31
0
80
60
40
20
0
100
Relative Abundance100
80
60
40
20
0
85.18
99.08
275.06
127.08 293.13
148.98
141.06
166.99
148.92
166.93
307.14
DiNP
DiDP
275.09 429.27
100 150 200 250 300 350 400 450
m/z
NL: 6.90E6
VegasSmoothieCup_AllOpt_201
21004#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_201
21004#644-666 RT: 1.92-1.95
AV: 2 SB: 3 0.04-0.15 F: ITMS
+ c NSI Full ms2 447.10
21

Coffee Stirrer
100
80
60
40
20
0
80
60
40
20
0
100
Relative Abundance100
80
60
40
20
0
57.12
85.12
148.95
148.96
205.02
113.09 261.03
166.96
127.05
148.94
279.18
279.11
275.05
293.11
373.27
363.18
DiBP
DEHP
391.32
DiNP
50 100 150 200 250 300 350 400
m/z
418.36
NL: 6.05E4
AircraftCoffeeStick_AllOpt_20121005#12
4-156 RT: 0.41-0.46 AV: 3 SB: 8
2.52-2.78 F: ITMS + c NSI Full ms2
279.10@hcd10.00 [50.00-290.00]
NL: 9.71E4
AircraftCoffeeStick_AllOpt_20121005#12
8-164 RT: 0.43-0.48 AV: 3 SB: 5
0.04-0.21 F: ITMS + c NSI Full ms2
391.20@cid20.00 [75.00-400.00]
NL: 6.26E4
AircraftCoffeeStick_AllOpt_20121005#12
4-232 RT: 0.40-0.68 AV: 10 SB: 5
0.06-0.20 F: ITMS + c NSI Full ms2
419.30@cid25.00 [80.00-425.00]
22

Airline Plastic Fork: DEHP & DnOP
AircraftFork_AllOpt_20121005 10/5/2012 8:53:06 AM Aircraft utensil packet
250C, He, tweezer
DnOP
RT: 1.25 - 1.93
100 NL: 1.03E6
m/z= 258.79-263.10 F: ITMS
+ c NSI Full ms2 391.10 MS
50
AircraftFork_AllOpt_20121005
100 0
50
100 0
50
More heat into the sample as it stays in heated gas
DEHP
DEHP
0
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)
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
Relative Abundance
100
50
0
100
50
0
100
50
0
148.97
80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400
m/z
261.10
113.11 279.14
261.08
148.98
391.31
279.14
113.11 167.01
391.35
113.09
148.96 279.14
261.05
166.97
DnOP
DnOP DEHP
Mix
DEHP
247.15 275.18
355.35
373.37
391.36
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
23

Yoga Mat
YogaMat_AllOpt_20121005 10/5/2012 11:01:09 AM Katarina's yoga mat
250C, He, tweezer
Blue
Black
RT: 0.00 - 1.62
100 NL: 6.60E7
m/z=
50
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
100 0
YogaMat_AllOpt_20121005
NL: 4.63E6
m/z= 260.50-261.50+278.50-279.50 F: ITMS + c NSI Full ms2
50
391.10 MS YogaMat_AllOpt_20121005
100 0
50
100 0
50
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
Relative Abundance
100
0
100
0
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6
Time (min)
50
50
0
100
50
0
100
50
0
61.17
86.08
50 100 150 200 250 300 350 400 450 500
m/z
391.26
149.00 279.17
134.08 177.09 195.09 228.17 267.17 313.17 335.25 379.25 419.34 447.34 466.34
148.96
279.11
113.10 166.97
261.03
99.12
195.11 257.23 285.31 331.30 373.28
391.33
275.07
127.06 293.13
85.14 148.95 401.30
115.15
195.09 239.14
313.34 363.17
419.39
289.07 307.17
141.07
429.34
447.41
166.96
99.13 127.06 195.10
255.21
341.25 387.30
85.21 239.14
Full Scan
DEHP
DiNP
DiDP
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
24

Summary
• 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
Email: crawford@ionsense.com
25