New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control
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1 New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Dr. Nuria Cortés-Francisco Servei de Química Laboratori Agència Salut Pública Barcelona Vancouver, August 22 nd, 217
2 Summary Laboratori Agència de Salut Pública de Barcelona Orbitrap technology Brominated Flame Retardants: PBDEs Polychloronaphtalenes Ndl-PCBs New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 2/26
3 Laboratori Agència de Salut Pública Food Safety Regulations Accreditation The official control labs should: - help to ensure food safety - fulfill regulations - work under accreditation ISO/IEC 1725 New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 3/26
4 Laboratori Agència de Salut Pública Regulations INSTRUMENTATION VALIDATION New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 4/26
5 Laboratori Agència de Salut Pública Food Safety Accreditation New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 5/26
6 Laboratori Agència de Salut Pública Method matrix matched standard calibration curve (MMSCC) SCC (Standard calibration curve): calibration using standards in solvent MSCC (Matrix matched standard calibration curve): addition of standards to sample extracts from blank matrix. MMSCC (Method matrix matched standard calibration curve): addition of standards to blank matrix prior to extraction. New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 6/26
7 Laboratori Agència de Salut Pública Method matrix matched standard calibration curve (MMSCC) Matrix effect evaluation: egg analysis Blue: SCC standards in solution Red: MSCC Matrix matched Green: MMSCC Matrix matched New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 7/26
8 Methods based on Orbitrap at LASPB Analytical methods already in use: ANALYSIS OF VETERINARY DRUGS: Antibiotics Hormones Chloramphenicol Beta agonists Nitrofurans Nonsteroidal anti-inflammatory drugs.. ANALYSIS OF MARINE BIOTOXINS ANALYSIS OF CONTAMINANTS FROM FOOD PACKAGING ANALYSIS OF PROCESS CONTAMINANTS IN FOODSTUFF New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 8/26
9 New instrumentation: Orbitrap GC-MS Q-Exactive Exactive * Installed 14/12/215 Brominated Flame Retardants: PBDEs Polychloronaphtalenes (PCNs) Ndl-Polychlorobiphenyls (ndl-pcbs) Thermo Scientific Q Exactive GC system Unprecedented Depth in Analysis RP 12, m/z 2) EI/CI; Full-scan, Timed-SIM MS/MS capability New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 9/26
10 Bringing GC and Orbitrap Technology Together C-TRAP AQT Quadrupole Bent Flatapole HCD Cell ExtractaBrite Ion Source Orbitrap Mass Analyzer * Courtesy of Thermo Fisher Scientific New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 1/26
11 Polybrominated diphenyl ethers (PBDEs) (a) Polybrominated diphenyl ethers (PBDEs) BDE 28 BDE 47 BDE 49 BDE 99 BDE 1 BDE 138 BDE 153 BDE 154 BDE 183 BDE 29 Brx O x=1-5, y=-5 Bry eggs and egg products milk and dairy products meat and meat products animal and vegetable fats and oils fish and other seafood products for specific nutritional uses food for infants and small children Analytical methods with a limit of quantification of,1 ng/g wet weight or lower 1 ppt in the sample New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
12 Polybrominated diphenyl ethers (PBDEs) 1 5 RT: 6.76 AA: BDE BDE-49 RT: 7.3 AA: RT: 7.21 AA: 6831 BDE-1 BDE-47 RT: 7.7 AA: RT: 7.8 AA: BDE-99 RT: 8.1 AA: RT: 8.44 AA: TUNA sample spiked with.1 ng/g for all the PBDEs BDE-154 BDE-153 BDE-138 RT: 8.67 AA: BDE RT: 1.29 AA: Time (min) BDE-29 New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
13 HRMS approach With the new HRMS instruments we are capable of acquiring the full isotopic pattern without losing sensitivity and working at R: 3, (FWHM, m/z 2) or higher C 12 O Br 4 81 Br ppm Quantification m/z 2.3 After cleanup, the extract is concentrated to 2 µl and labeled injection internal standards are added. An aliquot of the extract is injected into the gas chromatograph (GC). The analytes are separated by the GC and detected by a high-resolution (>5) mass spectrometer. Two exact m/z s are monitored at each level of bromination (LOB) throughout a predetermined retention time window C 12 O Br 5 81 Br ppm Confirmation m/z * C 12 O Br 3 81 Br ppm m/z New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
14 HRMS approach Quantification m/z C 12 O Br4 81 Br ppm MOLECULAR ION C12 O Br5 81 Br ppm C12 O Br5 81 Br3.229 ppm * C12 O Br3 81 Br5.553 ppm Confirmation m/z TUNA sample spiked before extraction at,1 ng/g with BDE m/z m/z New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
15 Optimization of EI Source Parameters 5 25 ev 4 ev 5 ev 7 ev Response BDE-28 BDE-49 BDE-47 BDE-1 BDE-99 BDE-154 BDE-153 BDE-138 BDE-183 BDE-29 New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
16 Acquisition modes.5 µg/l PBDE standard 1 5 RT: 7.32 AA: RT: 7.4 AA: Full Scan m/z RT: 1.42 AA: RT: 7.32 AA: RT: 7.41 AA: SIM 4 Da 1 5 RT: 1.43 AA: RT: 7.41 AA: RT: 1.43 AA: RT: 7.32 AA: SIM 1 Da RT: 7.32 AA: 3474 RT: 7.41 AA: SIM 25 Da 1 5 RT: 1.46 AA: 3416 RT: 1.42 AA: RT: 7.32 AA: RT: 7.4 AA: SIM 5 Da 1 5 RT: 1.42 AA: Time (min) Time (min) New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
17 Acquisition modes TUNA sample spiked with,1 ng/g 1 RT: 6.86 AA: Full Scan m/z RT: 6.87 AA: 3316 SIM 5 Da RT: 7.31 AA: 1988 RT: 7.39 AA: RT: 7.31 AA: RT: 7.4 AA: RT: 7.79 AA: RT: 7.8 AA: RT: 8.2 AA: 2467 RT: 1.45 AA: RT: 8.2 AA: RT: 8.19 AA: RT: 8.54 AA: RT: 8.77 AA: 3263 RT: 1.44 AA: RT: 8.2 AA: RT: 8.54 AA: RT: 8.78 AA: RT: 1.45 AA: RT: 1.46 AA: Time (min) Time (min) New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
18 Acquisition modes TUNA sample spiked with,1 ng/g 1 RT: 6.86 AA: Full Scan m/z RT: 6.87 AA: 3316 SIM 5 Da 5 BDE-28 5 BDE BDE-1 RT: 7.79 AA: 5166 BDE BDE-1 RT: 7.8 AA: RT: 7.91 AA: 1975 BDE-99 1 RT: 8.77 AA: RT: 8.78 AA: BDE BDE matrix co-elution BDE-29 RT: 1.45 AA: BDE-29 RT: 1.46 AA: Total Ion Chromatogram Total Ion Chromatogram Time (min) Time (min) New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
19 Acquisition modes TUNA sample spiked with,1 ng/g 1 AA: 5166 Full Scan m/z AA: SIM 5 Da Relative Abundance 1 Relative Abundance BDE-1 BDE-99 AA: 3263 BDE-183 Relative Abundance Relative Abundance BDE-1 AA: 1975 BDE-99 BDE-183 AA: 1253 Ion Time: 11 ms Ion Time: 119 ms C- trap saturation due to matrix is overcome using the Q! New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
20 Polybrominated diphenyl ethers (PBDEs) Monkfish extracted matrix-matched calibration curve.1 ng/g to.3 ng/g PBDEs BDE-29 Quantification m/z Confirmation m/z New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, 217 2/26
21 Polychloronaphtalenes (PCNs) Samples analyzed Fish and seafood Eggs and egg products Instrumental Analysis PTV injection HP-5MS, 3 m, i.d..25 mm,,25 µm GC column Optimized Tune parameters High Resolution ( 6, FWHM m/z 2) Wide SIM: 25 m/z window to monitor the whole isotopic pattern Mass accuracy < 1ppm External calibration Listed under Annex A and C with specific exemptions for use RT: SM: 7G RT: AA: Relative Abundance Relative Abundance Relative Abundance Relative Abundance Relative Abundance PCN-48 (tetra) RT: 15.5 AA: 2583 PCN-52 (penta) RT: 17. AA: 5512 PCN-73 (hepta) PCN-66/67 (hexa) RT: AA: PCN-75 (octa) RT: 2.89 AA: Time (min) New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
22 Polychloronaphtalenes (PCNs) If m=4 and n=4 PCN-75: Octachloronaphthalene LOQ=,1 ng/g wet weight Quantification m/z Confirmation m/z New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
23 Non-dl-Polychlorobiphenyls (n-dl-pcbs) Analytical requirements for the official control of the levels of non-dioxin-like PCBs: PCB-18,8 ng/g wet weight,1 ng/g fat,25 ng/g wet weight,31 ng/g fat GC-MS/MS (QqQ) + MRM (393,7 -> 324,) D D Smooth + MRM (395,7 -> 326,) D D Smooth x1 2 29,346 29,321 min. min. x1 21 Ratio = 56,5 46, (1, (74, %) %) 1 2,87 29,344 * 29,327 min. min. 3,5,9 2,6 6,5 2,4,8 3 2,2 6 5,5 2 2,5,7 1,8 5,6 1,6 2 1,4 4,5,5 1,5 1,24,4 3,5 1 1,8,3,63,5,2,4 2,5,2,1 2 28,9 28, ,1 29,129,2 29,29,3 29,3 29,4 29,4 29,5 29,5 29,6 29,6 29,7 29,7 29,8 28,9 28, ,1 29,129,2 29,29,3 29,3 29,4 29,4 29,5 29,5 29,6 29,6 29,7 29,7 29,8 Acquisition Time (min) Acquisition Time (min) New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
24 Ndl-Polychlorobiphenyls (Ndl-PCBs) Analytical requirements for the official control of the levels of non-dioxin-like PCBs: PCB-18 Full Scan SIM PRM PCB-28 Total Ion Chromatogram Meat sample spiked at.25 ng/g Total Ion Chromatogram Standard New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
25 Conclusions GC-Orbitrap is a useful tool for control labs: outstanding sensitivity high selectivity when analyzing complex matrices high robustness helps to fulfill new Regulations with extremely low limits (e.g. PBDEs) helps to confirm doubtful results But some expertise and optimization of instrumental parameters is needed. New Methodologies for POPs Analysis using Orbitrap GC-MS in Official Food Control Vancouver, August 22 nd, /26
26 Thanks for your attention Contact information:
SUPPORTING INFORMATION FOR: CONCENTRATIONS OF POLYBROMINATED DIPHENYL ETHERS, HEXABROMOCYCLODODECANES AND TETRABROMOBISPHENOL-A IN BREAST MILK FROM
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