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Guillaume ten Dam DSP‐Systems, Darwinstraat 7A, 6718XR Ede, The Netherlands Automation of the sample purification method for the analysis of all 209 polychlorinated biphenyls and dioxins
Guillaume ten DamDSP‐Systems, Darwinstraat 7A, 6718XR Ede, The Netherlands
Automation of the sample purification method for the analysis of all 209 polychlorinated biphenyls and dioxins
DioxinSamplePreparation
Wim Traag• Founder DSP‐Systems• Dioxin, PCB and POP analysis ≥ 40 years• Collaborator in numerous international projects on chemical analysis
Chris van Wakeren• Specialist in automated sample preparation ≥ 10 years• Sales and technical expert
Guillaume ten Dam• Dioxin, PCB and POP analysis ≥ 10 years• Participated in EU workgroups
Automation of the sample purification method for the analysis of all 209 polychlorinated biphenyls and dioxins
Markesteijn J., Thermo Fisher Scientific user meeting at Dioxin2019Are You Being Served? The Benefits of DualData Acquisition in a Routine Dioxin Lab
ten dam G. et al., Journal of Chromatography A, Volume 1477, 16 December 2016, Pages 76‐90The performance of atmospheric pressure gas chromatography–tandem mass spectrometry compared to gas chromatography–high resolution mass spectrometry for the analysis of polychlorinated dioxins and polychlorinated biphenyls in food and feed samples
Marchand P. et al., Organohalogen Compounds Vol. 76, 546‐549 (2014)A new and highly innovative automatic purification system evaluated for dioxins and PCBs
Approach – Published and experimental
Hayward D. G. et al., Chemosphere 256 (2020) 127023New approach for removing co‐extracted lipids before mass spectrometry measurement of persistent of organic pollutants (POPs)in foods
Dioxin & no‐PCB fractionin approx. 1.5 ml toluene
mo‐PCB & marker PCB fractionin approx. 1.5 ml toluene Waste
approx. 90 ml hexane
Design ‐ Goal
EN 16215:2012
Recoveries Dioxin and DL‐PCB ‐ EN 16215:2012
0
50
100
150
0
50
100
150
Advantages
• Fractions of 1.5 ml in a GC‐vial
• Low solvent consumption (<100 ml)
• No Dichloromethane needed
• Purification finished within 80 min
• No cross‐contamination
• No exposure to chemicals
General cleanup
25.2 25.6 26.0 26.4 26.8 27.2 27.6
#138
#167 #1
56
#157
60˚C
25.2 25.6 26.0 26.4 26.8 27.2 27.6
TypeSize[mm]
Standard20φ
Mini18 φ
DXNmini18 φ
Sample Food 3 g fat
Environmental 5 g d.m.
Environmental5 g d.m.
PurificationAgNO3-Silica gel AgNO3-Silica gel AgNO3-Silica gelH2SO4-Silica gel H2SO4-Silica gel H2SO4-Silica gel
ConcentrationCarbon Carbon CarbonAlumina Alumina -
Elution
To waste 90mL Hexane 85mL Hexane 85mL HexaneDXN fraction 1.5 mL Toluene 1.5 mL 1.5 mL
PCB fraction 1.5 mL Toluene 1.5 mL
Run time 80 min 78 min 73 min
WHO Dioxin & DL‐PCB and marker PCB
ten Dam et al., NEMC 2020, A Next Generation Automated Purification System for the Analysis of Dioxins and Associated POPs
So what about all 209PCB?EPA method 1618
mo‐PCB & marker PCB fractionin approx. 1.5 ml toluene
Dioxin & 209PCB?
Dioxin & no‐PCB fractionin approx. 1.5 ml toluene
Recovery 13C DL‐PCB and 13C NDL‐PCB in samplesconventional 2 run analysis (blue) vs single run analysis (pink)
0
30
60
90
120
150
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
13CPCB77
13CPCB81
13CPCB126
13CPCB169
13CPCB114
13CPCB118
13CPCB123
13CPCB156
13CPCB157
13CPCB167
13CPCB189
13CPCB28
13CPCB52
13CPCB101
13CPCB138
13CPCB153
13CPCB180
Recovery %
ten Dam et al., DSP‐Systems Application note 2019:03, Performance study of the Determination of DL‐PCBs and NDL‐PCBs in one single GC‐HRMS measurement using a Miura GO‐xHT for Sample Purification (Part I; Food and Feed)
Similarity DL‐PCB and NDL‐PCBsingle run analysis (pink) vs conventional 2 run analysis (blue)
0
20
40
60
80
100
120
140
160
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
combine
d
sepa
rate
PCB77 PCB81 PCB126 PCB169 PCB105 PCB114 PCB118 PCB123 PCB156 PCB157 PCB167 PCB189 PCB28 PCB52 PCB101 PCB138 PCB153 PCB180
Similarity %
+1s
x̅
‐1s
ten Dam et al., DSP‐Systems Application note 2019:03, Performance study of the Determination of DL‐PCBs and NDL‐PCBs in one single GC‐HRMS measurement using a Miura GO‐xHT for Sample Purification (Part I; Food and Feed)
Experience with lower chlorinated congeners Polychlorinated naphthalene's
ten Dam et al., DSP‐Systems application note 2018:12, Feasibility study for the Determination of Polychlorinated Naphthalenes in Food and Feed samples
Alumina fractionin approx. 1.5 ml toluene
PCN’s
Carbon fractionin approx. 1.5 ml toluene
ten Dam et al., DSP‐Systems application note 2018:12, Feasibility study for the Determination of Polychlorinated Naphthalenes in Food and Feed samples
Experience with lower chlorinated congeners Polychlorinated naphthalene's
0
50
100
150
13CPCN27
13CPCN52
13CPCN64
13CPCN67
13CPCN73
13CPCN75
Recovery inyernal standards of PCN’s in samples
0
50
100
150
PCN2 PCN3 PCN5 PCN13 PCN24 PCN42 PCN46 PCN52 PCN53 PCN66 PCN68 PCN73 PCN75
Recovery (%
)
Blank Pig fat PFAD Fishoil
Recovery of native PCN’s in samples
ten Dam et al., DSP‐Systems application note 2018:12, Feasibility study for the Determination of Polychlorinated Naphthalenes in Food and Feed samples
0
20
40
60
80
100
120
140
PCB1
PCB3
PCB1
2PC
B21
PCB5
/8PC
B38
PCB1
3PC
B11
PCB6
PCB1
82/187
PCB1
54PC
B155
PCB1
99PC
B148
PCB1
78PC
B201
PCB1
45PC
B197
PCB3
5PC
B196
/203
PCB1
7PC
B176
PCB1
86PC
B177
PCB1
71PC
B170
/190
PCB3
0PC
B158
PCB1
81PC
B140
PCB3
2PC
B139
/149
PCB1
66PC
B133
PCB1
31/142
/165
PCB1
00PC
B99
PCB2
06PC
B134
PCB1
30PC
B95/12
1PC
B91
PCB2
05PC
B87/11
5PC
B110
PCB1
19PC
B73
PCB1
29PC
B128
PCB4
5PC
B77
PCB9
8/10
2PC
B153
/168
PCB1
12PC
B116
/117
/125
PCB5
8PC
B41/64
/68
PCB1
04PC
B90
PCB8
1PC
B71/72
PCB6
9PC
B180
/193
PCB9
3PC
B62
PCB5
3PC
B55
PCB7
8PC
B60
PCB7
0PC
B67
PCB5
6PC
B74
PCB2
0/33
PCB2
9PC
B65
PCB1
67PC
B109
PCB1
26PC
B124
PCB2
5PC
B82
PCB1
27PC
B26
PCB5
2PC
B209
PCB1
23PC
B105
Standard method 2:1‐3g
Recovery of PCBsStandard method
Alum
ina
Carbon
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20
40
60
80
100
120
140
Average (n=3) recovery applying standard method 2: 1‐3g
alumina carbon waste
0
20
40
60
80
100
120
140
PCB52 PCB106/118 PCB209 PCB31/28 PCB123
Standard method 2:1‐3g
alumina carbon waste
Breakthrough of PCB209
Recovery of PCBsStandard method
no‐PCB fraction and slow eluters
Majority of PCBsA fraction of PCB209
A number of lower chlorinated PCBs
no‐PCB fraction and slow eluters
Majority of PCBsA fraction of PCB209
A number of lower chlorinated PCBs
Enhance elution
Increase absorption strength
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60
80
100
120
140
Ethyl acetate modifier
alumina carbon
1ml ethyl acetate added to a 9ml hexane standard solution
Improved recovery lower chlorinated PCBs
0
20
40
60
80
100
120
140
Toluene modifier
alumina carbon waste
1ml modifier added to a 9ml hexane standard solution
Improved recovery lower chlorinated PCBs
TypeSize[mm]
Standard20φ
Mini18 φ
DXNmini18 φ
Sample Food 3 g fat
Environmental 5 g d.m.
Environmental5 g d.m.
PurificationAgNO3-Silica gel AgNO3-Silica gel AgNO3-Silica gelH2SO4-Silica gel H2SO4-Silica gel H2SO4-Silica gel
ConcentrationCarbon Carbon CarbonAlumina Alumina -
Elution
To waste 90mL Hexane 85mL Hexane 85mL HexaneDXN fraction 1.5 mL 1.5 mL Toluene 1.5 mL Toluene
PCB fraction 1.5 mL 1.5 mL Toluene -
Run time 80 min 78 min
209PCB18 φ
Environmental5 g d.m.
AgNO3-Silica gelH2SO4-Silica gel
Alumina TM--
N/A
-
WHO Dioxin & 209PCB – In development
+1ml Toluene
Dioxins
PCBs
Recovery of all 209PCBs
0
50
100
150
Sample bottle rinsed with 1ml toluene and this 1ml toluene added to a 18Ø column
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10
20
30
40
50
60
70
80
90
0 : 2.5 0.5 : 2 2 : 0.5
Reovery %
V toluene : V hexane
2378‐TCDD PCB3 PCB15 PCB81
Effect of toluene on recovery of PCBs and Dioxins
WasteCarbon
TypeSize[mm]
Standard20φ
Mini18 φ
DXNmini18 φ
Sample Food 3 g fat
Environmental 5 g d.m.
Environmental5 g d.m.
PurificationAgNO3-Silica gel AgNO3-Silica gel AgNO3-Silica gelH2SO4-Silica gel H2SO4-Silica gel H2SO4-Silica gel
ConcentrationCarbon Carbon CarbonAlumina Alumina -
Elution
To waste 90mL Hexane 85mL Hexane 85mL HexaneDXN fraction 1.5 mL 1.5 mL Toluene 1.5 mL Toluene
PCB fraction 1.5 mL 1.5 mL Toluene -
Run time 80 min 78 min
209PCB18 φ
Environmental5 g d.m.
AgNO3-Silica gelH2SO4-Silica gel
Alumina TM--
N/A
-
209PCB only – In development
PCBs
Stronger absorption
TM
O
AlOO
AlO
PCB molecule = Electron-donor
Electron-acceptor
d-orbit
TransitionMetalLewis acid point
Lacking electrons
Image of Adsorption Principle of PCB Molecules on Alumina
Improving absorption
Fujita et al. Dioxin2019
0
50
100
150
PCB209 57%
Recovery of PCBs from using a alumina oxide TM concentration column
Fujita et al. Dioxin2019
Fujita et al. Dioxin2019
10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.000
2.0+E6
4.0+E6
6.0+E6
8.0+E6 TIC: 5g of sunflower oil / Carbon fraction
10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.000
TIC: 5g of sunflower oil / Alumina fraction
2.0+E6
4.0+E6
6.0+E6
8.0+E6
10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.000
TIC: 5g of sunflower oil / Waste fraction
2.0+E6
4.0+E6
6.0+E6
8.0+E6
1.0mL
Waste
1.2mL
DXN fraction
PCB fraction
Advantages of fractionation
Presented by Guillaume ten Dam| [email protected] | www.DSPsystems.eu
Questions?
• 2019:11 ‐ Comparison of available MS Systems for the Analysis of Dioxins and PCBs
• 2019:03 ‐ Single injection analysis of PCBs and Dioxins in food and feed using GO‐HT sample purification
• 2018:12 ‐ Feasibility study on Polychlorinated Naphthalenes(PCNs) analysis using GO‐HT sample purification
• 2017: Simultaneous Analysis for Dioxins, PCBS and PBDES Part III
• 2016: Simultaneous Analysis of Dioxins, PCBs and PBDEs Part II
• 2015: Simultaneous Analysis of Dioxins, PCBs and PBDEs Part I
• Hayward D. G. et al, Chemosphere Volume 256, October 2020, 127023, New approach for removing co‐extracted lipids before mass spectrometry measurement of persistent organic pollutants (POPs) in foods
• Fujita et al. OrganohalogenCompounds (2016), Simulteneousanalysis of dioxins, PCBs, and PBDEswith a fully automated sample preparation system (II: validation)
• Marchand P. et al., OrganohalogenCompounds Vol. 76, 546‐549 (2014), A new highly innovative automatic purification system evaluated for Dioxins and PCBs.
