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#

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Conc'n detectedRunning MeanRunning Mean + 2s.