evolution of an analytical method for brominated flame retardants k.a. macpherson, t.m. kolic and e....

Evolution of an Analytical Method for

Brominated Flame Retardants

K.A. MacPherson, T.M. Kolic and

E. J. Reiner

Ministry of the Environment –Toronto, Ontario

2

Overview

• Ontario Ministry of Environment– Dioxin Laboratory

• BFRs• Sample Preparation • GC-HRMS method development (2000 – 2006)• Current status/capability• Future analyses / Method Development

3

MOE - Laboratory Services Branch

LaSB provides analytical support for the ministry's – environmental monitoring – regulatory programs– data supports standard setting– analytical method development (emerging issues)– ensures the data quality of ministry compliance

enforcement and emergency analytical testing– Emergency response

4

MOE – LaSB -Dioxin LaboratoryPolychlorinated Dioxins and Furans (DFPCB - E3418)

Dioxin-Like PCBs (DFPCB - E3418)

Congener Specific PCBs by HRMS (PCBC - E3459)

Brominated Flame Retardants (PBDE - E3430)

Polychlorinated Naphthalenes (PCN - E3431)

Perflourooctane Sulfonate (PFOS – E3457)

Brominated and mixed Br/Cl dioxins and furans (PBDDc/PBDFs)*

Polychlorinated Diphenylethers (PCDE - 3432)*

LSB Method numbers in brackets (*) = Method under development

5

BFRs in the Environment

• Initial estimates by Environment Canada suggest:

– sewage biosolids may be a major contributor of the total PBDE loadings to the environment (excluding landfills)

– Sewage sludge, a product from waste water treatment process, which is organic rich and often applied as fertilizer on agricultural land

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BFR survey in Biosolids

• NMA – Nutrient Management Act

• 2002 Legislation – The purpose of this Act is to provide for the management of

materials containing nutrients in ways that will enhance protection of the natural environment and provide a sustainable future for agricultural operations and rural development.

• In response to increased tendency for corporate farming, diminishing concern for land stewardship and increased concern for water quality & quantity

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Sam

ple P

reparatio

n

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Sample Cleanup Considerations

• Acid/Base/AgNO3 cleanup may be repeated with difficult samples

• Standard dilution for injection on GC-HRMS =1:50

• Difficult samples (& “overloaded”) may require dilutions up

= 1:800 (many co-extractables present)

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Ideally one injection per sample

• Major & Minor congener consideration

– Some yet un-identified congeners present at minor concentrations

– Some predominant congeners need dilution for accurate quantification

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The Era Begins….2000

• 4 function - SIM Experiment• HP 6890+ GC, 40m .18mm x .18µm rtx500 column• Ultima NT, running MassLynx (v4.0) software

• Monitor & quantify six congeners;

Br3 – Br7 : BZ# 28,47,99,100,153,154

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2002 – Search for Deca • 6 function - SIM Experiment

• Monitor & quantify;

Br1 – Br10 (no Br8/ Br9)

• 13C12-BDE analytical standards available in calibration standard:

BZ#: 3, 15, 28, 47, 99, 153, 154, 183, 209

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2004 - 2006

• Non-bde analytical standards available in CVS– HBB – Hexabromobenzene– PBEB – Pentabromoethylbenzene– BB-153 – Hexabromobiphenyl– BTBPE – 1,2-Bis(2,4,6-Tribromophenoxyethane)– DBDPE - Decabromodiphenylethane

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Analytical Challenges

• Ensure Deca BDE efficiently introduced to GC

– Injector conditions

– Temperature

– Liner

– Carrier gas flow• Ensure Deca BDE efficiently leaves GC!

– Minimal Film Thickness

– Minimal length

– Appreciable ID

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2000-2006

• Uniliner emulates on-column injection and reduces discrimination

• Uniliner potentially effective without sealing column and would also increase longevity

• Minimal injections no matter which injection port liner when high concentrations present

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Injector Maintenance !

• Over time non-volatile sample residues in injector liner provide sites for absorption of critical analytes

• Enhanced Response

With clean injector

(The Reporter On-Line)

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30m RTX500 .25mm x .15µm– 42 minutes

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2003

• SEPARATION • extract injected on each of 2 columns: 15m & 30m

• QUANTIFICATION -

• Br3 -Br7

30m x 0.25mm x 0.10µm DB5-HT

• Br10

15m x 0.25mm x 0.10µm DB5-HT

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GC experiments

15M

• Injector:280C• Split/Splitless• .9mL/min

• 110C• 40C/min 200 C• 10C/min 330 C

hold until deca elutes

30M

• Injector 280C• Split/Splitless• 1.8mL/min

• 110C• 10C/min 140 C• 80C/min 220 C• 5 C/min 330C

hold until deca elutes

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30m DB-5HT (0.25mm x 0.10µm)

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15m DB5-HT (0.25mm x 0.10µm)

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15m DB-5HT (0.25mm x 0.10µm) 2005 CVS

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Wide Range in Physical Properties

• Mass– 230 amu - mono– 972 amu - deca

• Melting Point– 300 C

• Boiling Point– 420 C

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Analytical Challenges - HRMS Experiment Calibration

• Perfluorokerosene (PFK) is used as a reference calibrant

• Instrument calibration requires PFK masses which bracket the range of ions in the ion window (SIR experiment).

• Originally, modification of PFK reference file (Ultima-NT) necessary for successful SIR calibration up to ~971 Da. (13C12BDE 209)

Filename=c:\masslynx\ref\pfk.ref

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Reference Compound Mixture• Perfluorokerosene-H (PFK)

• Boiling Point : 210 - 240C

• Supplier:

1 Industrial Dr., Pelham, NH 03076

• Catalogue No.: 16596

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Molecular Ion Quantification

WHY

• 13C12BDE 209 AvailableAnalytical Standard

• Greater probability of positive identification

• No interferents in native ion channel

WHY NOT

1. Better response for [M - 2Br] + ion

2. Difficult calibration at this mass range

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You never know what you’ll find

13C12-BDE 209

DBDPEDecabromodiphenylethane

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Leading to production of analytical standard

• “Characterization of Mass-Labeled [13C14]-Decabromodiphenylethane and its use as a Surrogate Standard in the Analysis of Sewage Sludge Samples”

A.Konstantinova*, G.Arsenaulta, B.Chittima, K.MacPhersonb, A.McAleesa, R.McCrindlec, D.Pottera, E.Reinerb, C.Tashiroa and B.Yeoa.

Chemosphere 2004

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2006 : Analysis for BFRs

• 1 Analytical Run GC: 15m DB5-HT (0.25mm x 0.10µm)

HRMS: 6 function SIM Experiment

CONGENERS: 46 Native + 5 non-BDE BFRs

19 13C12- BDEs4 Injection Standards

• ~16 min. for last elutor (BDE 209)• >900 samples reported

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Future Method Development

• Transfer method to benchtop GC-MS/MS

• Excellent sensitivity 1pg DecaBDE (molecular ion)

• Less cost per analysisSkill level for operation < sector instruments

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Jan25_05_c122767_0002a

14.80 15.00 15.20 15.40 15.60 15.80 16.00 16.20 16.40 16.60 16.80 17.00 17.20Time0

100

%

0

100

%

Jan25_05_c122767_0002a 6: Voltage SIR 15 Channels EI+ 959.1679

1.08e615.72

Jan25_05_c122767_0002a 6: Voltage SIR 15 Channels EI+ 969.2102

3.38e516.66

15.70

c122767-0002c 1/50

Time15.00 15.20 15.40 15.60 15.80 16.00 16.20 16.40 16.60 16.80

%

0

100

15.00 15.20 15.40 15.60 15.80 16.00 16.20 16.40 16.60 16.80

%

0

100

15.00 15.20 15.40 15.60 15.80 16.00 16.20 16.40 16.60 16.80

%

0

100

15.00 15.20 15.40 15.60 15.80 16.00 16.20 16.40 16.60 16.80

%

0

100

14un06_bfr3 10: MRM of 7 Channels EI+ 959.2 > 799.3

1.23e515.57

14un06_bfr3 10: MRM of 7 Channels EI+ 969.2 > 809.4

1.54e415.55

16.4515.65

14un06_bfr3 10: MRM of 7 Channels EI+ 969.2 > 485.2

1.66e416.45

15.57

14un06_bfr3 10: MRM of 7 Channels EI+ 969.2 > 482.2

2.10e416.45

DBDPE13C12-BDE 209

BDE 209

BDE 209

13C12-BDE 209

DBDPE

DBDPE

M S/

M S

H R M S

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10pg DecaBDE / 20pg DBDPE

32

Acknowledgements

• Tereza Gobran – Sample Preparation

• Wellington Laboratories – Analytical Stds.