Polybrominated diphenyl ethers Polybrominated diphenyl ethers by HRGC/HRMSby HRGC/HRMS
Coreen HamiltonCoreen Hamilton
Brian FowlerBrian Fowler
Louis HavilandLouis Haviland
Axys Analytical Services, Ltd.Axys Analytical Services, Ltd.
PBDE - IntroductionPBDE - Introduction
Method OutlineMethod Outline• Samples are spiked with Samples are spiked with 1313CC1212-labeled PBDE -labeled PBDE
analogues;analogues;
• Solvent extraction procedure;Solvent extraction procedure;
• Liquid chromatography clean-up;Liquid chromatography clean-up;
• Analyzed by high-resolution GC/MS;Analyzed by high-resolution GC/MS;
Method FlowchartMethod Flowchart
SoxhletSoxhletOrOr
Soxhlet/Soxhlet/Dean StarkDean Stark
SoxhletSoxhlet SolventSolventSoxhletSoxhlet
Lipid Analysis (Y/N)Lipid Analysis (Y/N)
Sample ExtractionSample ExtractionSolidSolid TissueTissue, , BloodBlood XADXAD WaterWater
Liquid Chromatography Clean-upLiquid Chromatography Clean-up
Analysis by HR GC/MSAnalysis by HR GC/MS
MS Method OverviewMS Method Overview
Typical Calibration Concentrations (pg/Typical Calibration Concentrations (pg/L)L)
Analyte groupAnalyte group CS-1CS-1 CS-2CS-2 CS-3CS-3 CS-4CS-4 CS-5CS-5
NativesNatives 2525 7575 250250 750750 27502750
PBDE-209PBDE-209 175175 525525 17501750 52505250 1575015750
LabelledLabelled 100100 100100 100100 100100 100100
1313CC1212 PBDE 209 PBDE 209 700700 700700 700700 700700 700700
Calibration Analytes
Homologue GroupHomologue Group Target CongenersTarget Congeners SurrogatesSurrogates
Mono-PBDE’sMono-PBDE’s PBDE 2PBDE 2 PBDE 3PBDE 3
Di-PBDE’sDi-PBDE’s PBDE 8PBDE 8 PBDE 15PBDE 15
Tri-PBDE’sTri-PBDE’s PBDE 17, 28PBDE 17, 28 PBDE 28PBDE 28
Tetra-PBDE’sTetra-PBDE’s PBDE 47, 66, 75PBDE 47, 66, 75 PBDE 47, 66, 75PBDE 47, 66, 75
Penta-PBDE’sPenta-PBDE’s PBDE 85, 99, 100PBDE 85, 99, 100 PBDE 85, 99, 100PBDE 85, 99, 100
Hexa-PBDE’sHexa-PBDE’s PBDE 138, 153, 154PBDE 138, 153, 154 PBDE 138, 153, 154PBDE 138, 153, 154
Hepta-PBDE’sHepta-PBDE’s PBDE 183, 190PBDE 183, 190 PBDE 183, 190PBDE 183, 190
Deca-PBDE’sDeca-PBDE’s PBDE 209PBDE 209 PBDE 209PBDE 209
Detection Limits
MATRIX-->MATRIX-->
Analyte:Analyte:
SolidSolid
pg/gpg/g
TissueTissue
pg/gpg/g
WaterWater
pg/Lpg/L
AirAir
pg/pg/samplesample
ProceduralProceduralBlankBlankLevelLevel
pg/sample pg/sample
Mono-PBDEMono-PBDE 1010 1010 100100 100100 <2000<2000
Di- to Penta-PBDEDi- to Penta-PBDE 22 22 2020 2020 <50<50
Hexa-PBDEHexa-PBDE 22 22 2020 2020 <100<100
Hepta-PBDEHepta-PBDE 55 55 5050 5050 <200<200
Nona-PBDENona-PBDE 1010 1010 100100 100100 <500<500
Deca-PBDEDeca-PBDE 1010 1010 100100 100100 <1000<1000
Sample SizeSample Size 10g10g 10g10g 1.0L1.0L 1 smpl1 smpl
Final Vol. (Final Vol. (L)L) 2222 2222 2222 2222Matrix Spiking, pg (PBDE Matrix Spiking, pg (PBDE
209)209)2460 (17540)2460 (17540)
Quantification and Quantification and IdentificationIdentification
PARAMETERPARAMETER SpecificationSpecificationRETENTION TIME RETENTION TIME WINDOWWINDOW
Natives must elute within 3 seconds Natives must elute within 3 seconds of matching labelled surrogate. of matching labelled surrogate. Other natives must elute within 6 Other natives must elute within 6 seconds of predicted value from cal seconds of predicted value from cal runrun
RELATIVE RELATIVE ABUNDANCEABUNDANCE
Quant. And conf. Ions are within Quant. And conf. Ions are within 15% of theoretical values15% of theoretical values
PEAK CENTROIDSPEAK CENTROIDS Quant. And conf. Ions coincide Quant. And conf. Ions coincide within 2 secondswithin 2 seconds
Quantification and IdentificationQuantification and Identification
QC PARAMETERQC PARAMETER SpecificationSpecificationANALYSIS ANALYSIS DUPLICATEDUPLICATE
Difference in native concentrations Difference in native concentrations between duplicate samples <40%.between duplicate samples <40%.
PROCEDURAL PROCEDURAL BLANKBLANK Matrix Dependent (Previous Slide)Matrix Dependent (Previous Slide)
MATRIX SPIKE MATRIX SPIKE RECOVERYRECOVERY 50 – 150%50 – 150%
SURROGATE STD SURROGATE STD RECOVERYRECOVERY
40-130% for all labelled surrogates, 40-130% for all labelled surrogates, except except 1313CC1212 PBDE 209 (25-400%) PBDE 209 (25-400%)
RRF CONTINUING RRF CONTINUING CAL VERCAL VER
±35% of the actual for all natives ±35% of the actual for all natives ±50% of actual for all labelled ±50% of actual for all labelled surrogates except surrogates except 1313CC1212 PBDE PBDE
209209 (25 - 400%)(25 - 400%)
BRACKETING CALBRACKETING CAL <25% difference between opening <25% difference between opening and closing cal verand closing cal ver’’s s
CHROMATOGRAM CHROMATOGRAM QUALITYQUALITY
peak tailing ratio of peak tailing ratio of 1313C-DPE 77 C-DPE 77 peak (baseline peak width back peak (baseline peak width back half:front half) <3 half:front half) <3
Quantification and Quantification and IdentificationIdentification
Instrument SensitivityInstrument Sensitivity
INSTRUMENT INSTRUMENT SENSITIVITYSENSITIVITY
S:N > 3:1 for 7.5 pg of mono S:N > 3:1 for 7.5 pg of mono –– hepta hepta BDPE and 52 pg pf PBDE 209 BDPE and 52 pg pf PBDE 209 injected, with ion ratios within injected, with ion ratios within specification. specification.
INSTRUMENT INSTRUMENT LINEARITYLINEARITY
RSD of the RRFRSD of the RRF’’s <25% for all s <25% for all native and labelled compounds native and labelled compounds except except 1313CC1212 PBDE 209 PBDE 209 < 100%.< 100%.
GC/MS Operating ConditionsGC/MS Operating Conditions
Source temp:Source temp: 300300CC Injector Injector Temp:Temp: 300300CC
Electron Electron EnergyEnergy 35 eV35 eV Injector:Injector:
Split/splitless Split/splitless at 300at 300C; 2 C; 2 min splitlessmin splitless
Trap Current:Trap Current: VariableVariable Injection Injection VolumeVolume 1.0 1.0 µµLL
Mass Mass ResolutionResolution
Final Sample Final Sample VolumeVolume 22 22 µµLL
Carrier GasCarrier Gas 5000 5000 Detector Detector Voltage:Voltage: 350-370 eV 350-370 eV
Temperature Temperature Program:Program:
100100C, hold for 3 min., 5CC, hold for 3 min., 5C/min. to 320/min. to 320C, C, hold for 5 min. Total run time 52 min.hold for 5 min. Total run time 52 min.
• All homologues analyzed in a single run;All homologues analyzed in a single run;• Issues of PBDE-209 breakdown in Issues of PBDE-209 breakdown in DB-5 DB-5
columncolumn (30m) (30m);;• Shorter column showed no improvement;Shorter column showed no improvement;• Use of parent ions to reduce interference Use of parent ions to reduce interference
from PCBfrom PCB’’ss
GC/MS GC/MS Conditions/ConsiderationsConditions/Considerations
Advantages/ChallengesAdvantages/Challenges
• Stability issues with PBDE-209;Stability issues with PBDE-209;
• Solubility of standards in weaker solvent Solubility of standards in weaker solvent (nonane vs. toluene);(nonane vs. toluene);
• Linearity of 209;Linearity of 209;
Summary of Detectable Levels in Blank Samples for PBDE-47 (n=19)
0.00
0.20
0.40
0.60
0.80
1.00
1.20
WG#
Co
nc'
n D
ete
cte
d
(ng
/Sa
mp
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Conc'n detectedRunning MeanRunning Mean + 2s.