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

6

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

7

Sam

ple P

reparatio

n

8

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)

9

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

10

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

11

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

12

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

13

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

14

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

15

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)

16

30m RTX500 .25mm x .15µm– 42 minutes

17

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

19

30m DB-5HT (0.25mm x 0.10µm)

20

15m DB5-HT (0.25mm x 0.10µm)

21

15m DB-5HT (0.25mm x 0.10µm) 2005 CVS

22

Wide Range in Physical Properties

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

• Melting Point– 300 C

• Boiling Point– 420 C

23

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

24

Reference Compound Mixture• Perfluorokerosene-H (PFK)

• Boiling Point : 210 - 240C

• Supplier:

1 Industrial Dr., Pelham, NH 03076

• Catalogue No.: 16596

25

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

26

You never know what you’ll find

13C12-BDE 209

DBDPEDecabromodiphenylethane

27

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

28

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

30

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

31

10pg DecaBDE / 20pg DBDPE

32

Acknowledgements

• Tereza Gobran – Sample Preparation

• Wellington Laboratories – Analytical Stds.