New simple and fast GC-MS/MS method for the...
1
CONffIDENCE: Contaminants in food and feed: Inexpensive detection for control of exposure The research leading to these results has received funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 211326. New simple and fast GC-MS/MS method for the simultaneous analysis of various groups of organohalogen pollutants and polycyclic aromatic hydrocarbons Kamila KALACHOVA Jana PULKRABOVA Lucie DRABOVA Michal STUPAK Jana HAJSLOVA * Department of Food Chemistry and Analysis, Institute of Chemical Technology, Prague, Technicka 3, Prague 6, Czech Republic * [email protected] Work package WP1a – Persistent Organic Pollutants (POP) WP leader: Jana Hajslova (ICT Prague) WP deputy: Willam Haasnoot (RIKILT) Introduction____________________________________________________________ Aim of the study____________________ Tested matrices_______________________ Target analytes_______________________ Sample preparation procedure_____________________________________________ Conclusions & future plans_____________ GC-MS/MS analysis_____________________________________________________ Acknowledgement____________________ Since the number of environmental contaminants which undergo the legislative control or are included in the monitoring programs of the European Food Safety Authority (EFSA) and other international bodies still increases the demand for high throughput, selective, sensitive and non–expensive analytical methods arises as well. Gas chromatography coupled to triple quadrupole mass spectrometry (GC–MS/MS) represents a powerful tool for highly sensitive and selective determination of various groups of persistent organic polutants (POPs) as well as for polycyclic aromatic hydrocarbons (PAHs). With regards to similarities in physico-chemical properties of these chemicals some steps in various “traditional” analytical methods are almost identical. Nevertheless, until now, any uniform analytical flow-chart encompassing all these target analytes has not been introduced into a routine practice. Fresh fish mussel tissue: Trout (Salmon trutta) – 1.6% fat Salmon (Salmon salar) –13.7% fat Salmon trutta To implement gas chromatography coupled to tandem mass spectrometry (GC–MS/MS) with triple quadrupole ion analyzer for the simultaneous analysis of several groups of emerging POPs and PAHs in fish tissue. To achieve low limits of quantifications (LOQs) to obtain relevant data for exposition studies which require quantification of target contaminants in food even at very low levels. To extend the list of target analytes defined in the CONffIDENCE project (includes several PCBs, PBDEs and PAHs) to involve also other emerging contaminants (e.g., methylated analogues of PAHs, organochlorinated pesticides (OCPs) and other BFRs) which are recommended for the monitoring not only by EFSA. O Br x Br y PBDE x + y = 1 to 10 O O Br x Br y PBDE x + y = 1 to 10 Cl x Cl y PCB x + y = 1 to 10 Cl x Cl y PCB x + y = 1 to 10 This research was supported by grant from the European research project CONffIDENCE “Contaminants in food and feed: Inexpensive detection for control of exposure” which is a part of Seventh framework program (call KBBE–2007–2–4–02) and by financial Support from Specific University Research (MSMT NO. 21/2010). 18 polychlorinated biphenyls (PCBs) 22 organochlorinated pesticides (OCPs) 25 brominated flame retardants (BFRs) 33 polycyclic aromatic hydrocarbons (PAHs) Extraction Water & ethyl acetate Partition NaCl & MgSO 4 Clean-up Silica minicolumn Elution profiles of target analytes Note: Recoveries of all target analytes after clean-up were higher than 75% except for dieldrine, endrine, β-endosulfane, and endosulfan sulfate which were irreversibly fasten on the silica. Benz(a)anthracene Benzo(a)pyrene HBCD Chrysene Thermo Scientific TSQ Quantum XLS triple quadrupole Column: Rxi-17Sil-ms (30m × 0.25mm × 0.25μm) Injection: PTV splitless (1μL) Oven temperature: 80°C (2min), @30°C/min to 240°C, @ 10°C/min to 340°C (10min) Carrier gas: helium (1.3 mL/min) Source temperature: 250°C Emission current: 50μA Time (min) 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 0 10 20 30 40 50 60 70 80 90 100 Relative Abundance 52 101 SRM 255.860 > 185.901 SRM 291.900 > 221.899 SRM 323.800 > 253.901 SRM 357.840 > 287.881 SRM 391.810 > 321.839 28 81 77 123 118 153 114 105 138 126 167 156 180 157 169 189 PCBs 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 Time (min) 0 10 20 30 40 50 60 70 80 90 100 Relative Abundance SRM 128.000 > 76.999 SRM 141.000 > 114.999 SRM 152.000 > 101.999 SRM 153.000 > 125.999 SRM 165.000 > 138.999 SRM 178.000 > 151.999 SRM 192.000 > 151.999 SRM 202.000 > 164.999 1MeNa Acy Na Ace Fln 2MeNa Phe Ant 1MePhe 2MeAnt Flt Pyr PAHs An example of chromatograms of PCBs and several PAHs and their methylated analogues – GC – MS/MS (EI). Table I: Comparison of LOQs achieved using different instrumental techniques. (N/A – not available) Within this study a sample preparation method previously developed for the determination of PCBs, PBDEs and PAHs in fish tissues was enlarged by other BFRs, OCPs, PAHs and their methylated analogues. The irreversible adsorption of dieldrine, endrine, β-endosulfane, and endosulfan sulfate on silica minicolumn was observed. GC-MS/MS (EI) was shown to be an effective tool for identification and quantification of all target analytes even at very low levels. The LOQs were in the range of 0.005-0.1 μg/kg. The higher values were reached for BDE 183 and some OCPs. The poor sensitivity was also observed in the case of octa-, nona- and decaBDE and decabromodiphenyl ethane for which the CI and shorter GC capillary column (15 m) is recommended. 0 10 20 30 40 50 60 70 80 90 100 [%] PCB101 PCB81 PCB77 PCB123 PCB114 PCB153 PCB105 PCB138 PCB126 PCB167 PCB156 PCB157 PCB180 PCB169 PCB189 BDE28 BDE49 BDE47 BDE66 BDE100 BDE99 BDE85 BDE154 BDE153 BDE183 PBT PBEB HBB BTBPE B[c]Fln CP[cd]P B[a]A Chr 5MeChr B[b]F+B[j]F+B[k]F B[a]P I[cd]P DB[ah]A B[ghi]P DB[al]P DB[ae]P DB[ai]P DB[ah]P 1MeNaph 2MeNaph DBT 1MePhe 2MeAnt 1MePyr 1MeChr 3MeChr Ace Acy Fln Phe Ant Flt Pyr
Transcript of New simple and fast GC-MS/MS method for the...
CONffIDENCE: Contaminants in food and feed: Inexpensive detection for control of exposure
The research leading to these results has received funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 211326.
New simple and fast GC-MS/MS method for the simultaneous analysis of various groups of organohalogen pollutants and polycyclic aromatic hydrocarbons Kamila KALACHOVA Jana PULKRABOVA Lucie DRABOVA Michal STUPAK Jana HAJSLOVA *
Department of Food Chemistry and Analysis, Institute of Chemical Technology, Prague, Technicka 3, Prague 6, Czech Republic* [email protected]
Work package WP1a – Persistent Organic Pollutants (POP) WP leader: Jana Hajslova (ICT Prague) WP deputy: Willam Haasnoot (RIKILT)
Introduction____________________________________________________________
Aim of the study____________________ Tested matrices_______________________
Target analytes_______________________
Sample preparation procedure_____________________________________________
Conclusions & future plans_____________
GC-MS/MS analysis_____________________________________________________
Acknowledgement____________________
Since the number of environmental contaminants which undergo the legislative control or are included in the monitoring programs of the European Food SafetyAuthority (EFSA) and other international bodies still increases the demand for high throughput, selective, sensitive and non–expensive analytical methods arisesas well.
Gas chromatography coupled to triple quadrupole mass spectrometry (GC–MS/MS) represents a powerful tool for highly sensitive and selective determination ofvarious groups of persistent organic polutants (POPs) as well as for polycyclic aromatic hydrocarbons (PAHs).
With regards to similarities in physico-chemical properties of these chemicals some steps in various “traditional” analytical methods are almost identical.Nevertheless, until now, any uniform analytical flow-chart encompassing all these target analytes has not been introduced into a routine practice.
Fresh fish mussel tissue: Trout (Salmon trutta) – 1.6% fatSalmon (Salmon salar) –13.7% fat
Salmon trutta
To implement gas chromatography coupled to tandem mass spectrometry(GC–MS/MS) with triple quadrupole ion analyzer for the simultaneousanalysis of several groups of emerging POPs and PAHs in fish tissue.
To achieve low limits of quantifications (LOQs) to obtain relevant data forexposition studies which require quantification of target contaminants infood even at very low levels.
To extend the list of target analytes defined in the CONffIDENCE project(includes several PCBs, PBDEs and PAHs) to involve also other emergingcontaminants (e.g., methylated analogues of PAHs, organochlorinatedpesticides (OCPs) and other BFRs) which are recommended for themonitoring not only by EFSA.
O
Brx BryPBDE
x + y = 1 to 10
OO
Brx BryPBDE
x + y = 1 to 10
Clx ClyPCBx + y = 1 to 10
Clx ClyPCBx + y = 1 to 10
This research was supported by grant from the European research project CONffIDENCE “Contaminants infood and feed: Inexpensive detection for control of exposure” which is a part of Seventh framework program(call KBBE–2007–2–4–02) and by financial Support from Specific University Research (MSMTNO. 21/2010).
18 polychlorinated biphenyls (PCBs)
22 organochlorinated pesticides (OCPs) 25 brominated flame retardants (BFRs) 33 polycyclic aromatic hydrocarbons (PAHs)
ExtractionWater & ethyl acetate
PartitionNaCl & MgSO4
Clean-upSilica minicolumn
Elution profiles of target analytes
Note: Recoveries of all target analytes after clean-up were higher than 75% except for dieldrine, endrine, β-endosulfane, and
endosulfan sulfate which were irreversibly fasten on the silica.
Benz(a)anthracene
Benzo(a)pyrene
HBCD
Chrysene
Thermo Scientific TSQ Quantum XLS triple
quadrupole
Column: Rxi-17Sil-ms (30m × 0.25mm × 0.25µm) Injection: PTV splitless (1µL) Oven temperature: 80°C (2min), @30°C/min to 240°C,
@ 10°C/min to 340°C (10min) Carrier gas: helium (1.3 mL/min) Source temperature: 250°C Emission current: 50µA
Time (min)8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
52
101
SRM 255.860 > 185.901SRM 291.900 > 221.899SRM 323.800 > 253.901SRM 357.840 > 287.881SRM 391.810 > 321.839
28
8177
123
118153
114 105138
126
167156
180
157 169 189PCBs
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0Time (min)
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
SRM 128.000 > 76.999SRM 141.000 > 114.999SRM 152.000 > 101.999SRM 153.000 > 125.999SRM 165.000 > 138.999SRM 178.000 > 151.999SRM 192.000 > 151.999SRM 202.000 > 164.999
1MeNa AcyNa AceFln
2MeNa
Phe
Ant
1MePhe
2MeAnt
Flt
Pyr
PAHs
An example of chromatograms of PCBs
and several PAHs and their methylated analogues –
GC – MS/MS (EI).
Table I: Comparison of LOQs achieved using different instrumental techniques. (N/A – not available)
Within this study a sample preparation method previously developed for thedetermination of PCBs, PBDEs and PAHs in fish tissues was enlarged byother BFRs, OCPs, PAHs and their methylated analogues.
The irreversible adsorption of dieldrine, endrine, β-endosulfane, andendosulfan sulfate on silica minicolumn was observed.
GC-MS/MS (EI) was shown to be an effective tool for identification andquantification of all target analytes even at very low levels.
The LOQs were in the range of 0.005-0.1 µg/kg. The higher values werereached for BDE 183 and some OCPs. The poor sensitivity was also observedin the case of octa-, nona- and decaBDE and decabromodiphenyl ethane forwhich the CI and shorter GC capillary column (15 m) is recommended.
0
10
20
30
40
50
60
70
80
90
100
[%]
PCB101 PCB81 PCB77 PCB123
PCB114 PCB153 PCB105 PCB138
PCB126 PCB167 PCB156 PCB157
PCB180 PCB169 PCB189 BDE28
BDE49 BDE47 BDE66 BDE100
BDE99 BDE85 BDE154 BDE153
BDE183 PBT PBEB HBB
BTBPE B[c]Fln CP[cd]P B[a]A
Chr 5MeChr B[b]F+B[j]F+B[k]F B[a]P
I[cd]P DB[ah]A B[ghi]P DB[al]P
DB[ae]P DB[ai]P DB[ah]P 1MeNaph
2MeNaph DBT 1MePhe 2MeAnt
1MePyr 1MeChr 3MeChr Ace
Acy Fln Phe Ant
Flt Pyr
