CCQM-K102 Polybrominated diphenyl ethers in sediment...CCQM-K102_Final report.doc E-mail:...

CCQM-K102 E-mail: [email protected]

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EUROPEAN COMMISSION JOINT RESEARCH CENTRE Directorate F- Health, Consumers & Reference Materials (Geel) Reference Materials Unit

CCQM-K102

Polybrominated diphenyl ethers in sediment

"Track A" - Low polarity analytes in abiotic matrix

Final Report

Coordinating laboratory:

Marina Ricci*, Penka Shegunova, Patrick Conneely

European Commission, Joint Research Centre (JRC)

Directorate F- Health, Consumers & Reference Materials

Retieseweg 111, 2440 Geel, Belgium

CCQM-K102_Final report.doc E-mail: [email protected]

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With contributions from:

Roland Becker

Bundesanstalt für Materialforschung (BAM), Germany

Mauricio Maldonado Torres, Mariana Arce Osuna

Centro Nacional de Metrologia (CENAM), Mexico

Tang Po On, Lee Ho Man

Government Laboratory Hong Kong (GLHK), Hong Kong S.A.R.

Song-Yee Baek and Byungjoo Kim

Korea Research Institute of Standards and Science (KRISS), Republic of Korea

Christopher Hopley, Camilla Liscio, John Warren

LGC Limited (LGC), United Kingdom

Véronique Le Diouron, Sophie Lardy-Fontan, Béatrice Lalere

Laboratoire National de Métrologie et d'Essais (LNE), France

Shao Mingwu

National Institute of Metrology (NIM), China

John Kucklick

National Institute of Standards and Technology (NIST), United States

Veronica Vamathevan

National Measurement Institute Australia (NMIA), Australia

Shigetomo Matsuyama, Masahiko Numata

National Metrology Institute of Japan (NMIJ), Japan

Martin Brits, Laura Quinn, Maria Fernandes-Whaley

National Metrology Institute of South Africa (NMISA), South Africa

Ahmet Ceyhan Gören, Burcu Binici

National Metrology Institute of Turkey (UME), Turkey

Leonid Konopelko, Anatoli Krylov, Alena Mikheeva

D.I. Mendeleyev Institute for Metrology (VNIIM), Russia


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TABLE OF CONTENTS

1. INTRODUCTION ....................................................................................................... 4

2. STUDY ANALYTES .................................................................................................. 5

3. STUDY MATERIAL .................................................................................................. 6

3.1 Material preparation and initial characterisation ................................................... 6

3.2 Homogeneity study ................................................................................................ 7

3.3 Stability study ........................................................................................................ 7

4. SAMPLES DISTRIBUTION AND STUDY GUIDELINES ..................................... 9

5. CALIBRANTS' TRACEABILITY ........................................................................... 10

6. ANALYTICAL METHODS EMPLOYED .............................................................. 12

6.1 Dry mass determination ....................................................................................... 15

7. RESULTS .................................................................................................................. 15

8. APPROACHES TO UNCERTAINTY BUDGET ESTIMATION ........................... 17

9. KEY COMPARISON REFERENCE VALUES CALCULATION ......................... 21

10. DEGREES OF EQUIVALENCE (DOE) CALCULATION .................................... 24

11. CORE COMPETENCIES AND HOW FAR DOES THE LIGHT SHINE? ............ 30

12. CONCLUSIONS ....................................................................................................... 31

13. ACKNOWLEDGEMENTS ...................................................................................... 31

14. REFERENCES .......................................................................................................... 31

ANNEX A ......................................................................................................................... 32

ANNEX B ......................................................................................................................... 48

ANNEX C ......................................................................................................................... 52

ANNEX D ......................................................................................................................... 66

ANNEX E .......................................................................................................................... 68


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1. INTRODUCTION

Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants (i.e., to

reduce the inflammability) in many combustible commercial and household products,

such as polymers, electrical and electronic equipment, textiles, furniture, building and

packaging materials, since the last few decades. PBDEs are additives type brominated

flame retardants, meaning that they are not chemically bound but only physically

mixed/dissolved in the material. Due to the lack of covalent bonds between PBDEs and

the material, the release of these compounds into the environment can occur not only

when they are manufactured but also when products that contain them are used and

disposed of [1-2].

Environmental contamination by PBDEs has attracted public attention and concern in

recent years due to their large use, ubiquity (linked to the potential for long-range

atmospheric transport [3]) and high persistence, bioaccumulation and toxicity, thus

presenting a potential threat to wildlife and human health. Presence of PBDEs has been

reported in a range of environmental media and biota including fish, sediment, treated

sewage sludge and household dust [4-6]. PBDEs #28, 47, 99, 100, 153 and 154 are listed

as Priority Substances under the EU Water Framework Directive (WFD) and they are

also considered of primary interest for the environment in US and Canada (e.g., EPA

Method 1614).

Three technical mixtures namely pentabromodiphenyl ether (Penta-BDE),

octabromodiphenyl ether (Octa-BDE) and decabromodiphenyl ether (Deca-BDE) were

the most used brominated flame retardants until early 2000s, when the production and

usage of Penta– and Octa-BDE began to be regulated worldwide. The European Union

banned Penta-BDE and Octa-BDE in 2004 [7a] and prohibited the use of PBDEs (and

polybrominated biphenyls) in electric and electronic devices as of 1st July 2006 [7b].

The three commercial mixtures are composed of a mixture of congeners, and are named

according to their average bromine content, see Table 1.

Table 1. Composition of technical PBDEs products (% m/m) [8]

Technical

product tetraBDEs pentaBDEs hexaBDEs heptaBDEs octaBDEs nonaBDEs decaBDE

Penta-BDE 24-38 50-60 4-8

Octa-BDE 10-12 44 31-35 10-11 <1

Deca-BDE <3 97-98

The Track A suite of Key Comparisons was set by the CCQM OAWG to provide

objective evidences of core competencies needed to underpin the CMCs of an NMI for

the delivery of measurement services to customers. At the CCQM OAWG meeting held

in April 2011, it was agreed to have a comparison on brominated flame retardants in

sediment (fitting the category "low polarity analytes in abiotic matrix") as Track A study

for 2012 (service categories 5-10). This Key Comparison K102 was accompanied by the

parallel Pilot Study P138 (where the same study material was used).


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JRC-IRMM (renamed in 2016 as JRC, Directorate F - Health, Consumers & Reference

Materials) provided the sediment study material and coordinated the study, including

preparation and distribution of samples, data analysis and evaluation, drafting of reports.

NMIs/DIs with current published CMC claims in Appendix C of the Key Comparison

Database (KCDB) under the Mutual Recognition Arrangement of the International

Committee for Weights and Measures (CIPM MRA) or anticipating to propose CMC

claims in the selected measurement field (see Section 11) were expected to participate in

K102, so to avoid delays in the review, approval and acceptance of existing or future

CMC claims.

2. STUDY ANALYTES

Minimum reporting requirements for participants to CCQM-K102 were the mass

fractions (on a dry mass basis) of BDE 47, 99 and 153 in the freshwater sediment study

material. Possibility of reporting also the mass fraction of BDE 209 was left open to the

participants.

Among all PBDEs congeners, BDE 47, 99 and 153 were chosen considering the

following issues:

- analytical properties: different molar masses, different polarity and volatility, different

bromine substitutions (from tetra- to hexa-) (see Table 2)

Table 2. PBDEs selected as study analytes for CCQM K102

Congener Structural formula Chemical formula

(MW g/mol) Log Kow [9]

BDE-47

2,2',4,4'-tetraBDE

C12H6Br4O (485.8) 6.81 ± 0.08

BDE-99

2,2',4,4',5-pentaBDE

C12H5Br5O (564.7) 7.32 ± 0.14

BDE-153

2,2',4,4',5,5'-hexaBDE

C12H4Br6O (643.6) 7.90 ± 0.14

- analytical challenges: e.g. discrimination, likelihood of interferences. More specifically:

BDE 47: possible issues with discrimination in the GC injector

BDE 47, 99 and 153: possibility of co-elution with other environmental contaminants

O

Br Br

BrBr

O

Br Br

BrBr

Br

O

Br Br

BrBr

BrBr


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e.g., PCBs and/or other classes of flame retardants/halogenated compounds

- legislative and environmental relevance (see Section 1): BDE 47 and 99 are the most

abundant compounds present in the Penta-BDE commercial technical mixture

- attributes of the study material, with regard to the presence, mass fraction levels as well

as homogeneity and stability evaluation (see Section 3)

- connection to the previous pilot study CCQM-P114, Brominated flame retardants in

polypropylene, in which BDE 47 was measured.

The approximate mass fraction range on a dry mass basis of the selected PBDEs (47, 99

and 153) was anticipated to be between 1 and 50 μg/kg.

3. STUDY MATERIAL

3.1 Material preparation and initial characterisation

The sediment study material was prepared at JRC-IRMM. It originated from a river in

Belgium and contained PBDEs at levels typically found in environmental monitoring.

The material underwent a first series of processing steps: air-drying, sieving (1 mm), jet-

milling, homogenisation and γ-irradiation. The final powdered sediment was dispensed in

portions of about 40 g into 60 mL amber glass bottles with aluminium coated screw caps

sealed with a shrink film.

Water determination by volumetric Karl-Fischer titration was carried out in duplicate on

15 bottles randomly selected over the whole production batch and yielded a value of 0.45

± 0.06 g/100g (mean ± U, k = 2) (preliminary result with the drying-oven method: ≈ 0.5

g/100g). Total organic carbon measurements (in accordance to ISO 10694:1995) were

carried out in duplicate on 3 bottles, leading to a value of 0.641 ± 0.071 % C (mean ± U,

k = 2, dry mass basis).

The homogeneity data on this first batch prepared were unfortunately not satisfactory,

thus some bottles of the original batch were opened, re-sieved (125 µm), mixed, re-

bottled (see first batch) and subjected again to γ-irradiation. This re-processed second

batch of 92 units was further characterised with respect to the particle size. Particle size

analysis (laser light diffraction) was carried out in triplicate on one bottle randomly

chosen and revealed an average top particle size X90 = 55.64 ± 11.57 µm (mean ± U,

k=2). The study material was stored at + 4 °C ± 3 °C in the dark.

Fig. 1 Example of bottles of study material


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3.2 Homogeneity study

LNE kindly volunteered to analyse the newly prepared batch of study material by GC-

ID-MS/MS for assessing the homogeneity and stability of BDEs 28, 47, 99, 100, 153,

154, 183 and 209.

Homogeneity was assessed by triplicate analysis on 8 units (1 g sample intake) measured

in random order and under “quasi” repeatability conditions. Regression analyses did not

detect trends either in the analytical sequence or in the filling sequence. Estimation of

potential between-unit inhomogeneity (ubb) was accomplished by ANOVA. When the

mean square between groups (MSbetween) is smaller than the mean square within groups

(MSwithin), the between-unit variation sbb cannot be calculated and ubb*, maximum

inhomogeneity that could be hidden by method repeatability [10], is calculated instead.

The largest among sbb and ubb* is adopted as ubb, uncertainty contribution to account for

potential material inhomogeneity. The uncertainty contributions due to potential material

inhomogeneity were estimated as 2.7, 1.8 and 2.2 % for BDE 47, 99 and 153,

respectively, confirming the suitability of these analytes as study measurands for the

CCQM-K102 (Table 3).

Table 3. Homogeneity results (study analytes are green shaded)

% ubb

BDE 28 2.9 (ubb*)

BDE 47 2.7 (sbb)

BDE 99 1.8 (ubb*)

BDE 100 2.4 (ubb*)

BDE 153 2.2 (sbb)

BDE 154 2.9 (ubb*)

BDE 183 4.2 (sbb)

BDE 209 4.0 (sbb)

3.3 Stability study

An isochronous short-term stability study was performed at 18 °C with time points 0, 1, 2

and 4 weeks. One unit per time point was selected using a random stratified sampling

scheme and analysed in triplicate by GC-ID-MS/MS in random order and under “quasi”

repeatability conditions (1 g sample intake). For all PBDEs, the slopes of the regression

lines were not significantly different from zero (both at 95 % and 99 % confidence

levels), with the sole exception of BDE 209. For BDE 209, the slope of the linear

regression was significantly different from zero at 95 % (but not at 99 % confidence

level), possibly indicating degradation, and one outlier was detected (Grubbs single test

both at 99 and 95 % confidence level).

The stability study confirmed that the study measurands were stable at 18 °C for at least

4 weeks (Figure 2a, 2b and 2c). As a conservative mean, the dispatch and storage

temperature was set at 4 °C.


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Fig.2a Stability study at 18 ° C for BDE47

Fig.2b Stability study at 18 ° C for BDE99

Fig.2c Stability study at 18 ° C for BDE153


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4. SAMPLES DISTRIBUTION AND STUDY GUIDELINES

Fourteen NMIs/DI and one associate member subscribed for participation to the CCQM-

K102. Three bottles of the study material, each bottle containing 40 g of dried river

sediment, were dispatched with cooling elements by courier to the participants.

Additional bottles were sent to two laboratories upon request. The dispatch took place on

several dates during July and August 2014. Participants were asked to check the physical

conditions of the samples upon receipt, put it for storage at + 4 °C and report back to the

study coordinator. All laboratories received the samples in good conditions in 2-3 days,

except few laboratories for which delivery took longer due to customs problems (see

Table 4 for more details).

The study protocol, the registration form, the sample receipt form and the reporting sheet

for the results were sent to all laboratories by e-mail before or at the same time of the

dispatch of the samples. The Core Competency table was sent by e-mail in October 2014,

after revision by the CCQM OAWG at the fall meeting 2014.

Table 4. Samples dispatch schedule

Dispatch date Receipt date

Participants were requested to report the mass fractions (µg/kg) of BDEs 47, 99 and 153

on a dry mass basis in the study material applying their own analytical methodology, but

following some additional recommendations given by the study coordinator:

- upon receipt and until analysis, the samples should be stored at + 4 °C in the dark.

Equilibration to room temperature should be ensured before commencing the analytical

procedure. Before opening, the bottle should be shaken by turning upside down for 2

minutes for allowing material re-homogenisation.

The minimum sample intake had to be at least 1 g and the dry mass determination had to

be performed according the following protocol:

- a correction for dry mass shall be performed at the same time of the analysis by taking

2 separate portions of at least 1 g from each bottle analysed, drying them in an oven at

(105 ± 2) °C until constant mass is attained (subsequent weightings should not differ

more than 0.5 mg).


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The participants were requested to report the following data in the provided reporting

sheet to [email protected] (together with the Core Competency table) before the

deadline for submission (extended to 15th

April 2015):

Participant's details

Mass fractions (µg/kg) of each individual measurand (see Table 2) in the study

sample on a dry mass basis

Standard and expanded measurement uncertainties, with a detailed

description/breakdown of the full uncertainty budget

Description of the analytical procedure employed (extraction, clean-up,

separation/detection and quantification) as well as details concerning the

calibration and internal standards used (purity statement or verifications done at

the laboratory's premises etc…), especially if not mentioned in the Core

Competency table.

5. CALIBRANTS' TRACEABILITY

The information on the calibration standards used by the participants in CCQM-K102 are

given in Table 5.

CRM PBDEs solutions are available from NIST and NIM China. Pure PBDEs are also

commercially available from different suppliers as neat reagents (Chiron) and as

solutions (e.g., Chiron, Accustandard, Wellington Laboratories, CIL).

Almost half of the participating laboratories (6 out of 14) used NIST 2257 (PBDE

Congeners in 2,2,4-Trimethylpentane) as direct source of traceability. Other five

laboratories (NMISA, NMIA, LNE, CENAM and KRISS) used NIST 2257 in a transfer

value assignment process on commercially available standards (CIL, Wellington Lab.

and Accustandard); in particular, KRISS assessed the purity of the calibrants employed

by GC-FID, with cross-confirmation with SRM 2257; NMIA cross-checked the transfer

value assignment process by also using GBW(E)081124 and GBW(E)081125.

NIM used their own CRM, GBW(E)081124, Industrial penta-BDE in iso-octane. Two

laboratories, LGE and TUBITAK UME, assessed in-house the purity of the purchased

standards by means of qNMR, either on the neat substances or on solutions thereof.

mailto:[email protected]


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Table 5. Calibrants used in CCQM-K102

Participant

laboratory

Calibrants'

source

Purity % or

certified value

Purity assessment

JRC-IRMM NIST 2257

BDE47: 2.09±0.16 µg/g

BDE99: 2.127 ± 0.090 µg/g

BDE153: 2.048 ± 0.068 µg/g

N/A

LGC Chiron neat

substances > 98 %

qNMR to value assign BDE

solutions (+ verification on

neat BDE47 and 99)

NMISA CIL

BDE47: 68.28±5.77 µg/g

BDE99: 68.21±3.87 µg/g

BDE153: 68.59 ± 3.87 µg/g

values assigned using

SRM 2257

NIM GBW(E)081124

BDE47: 20.0 µg/mL U=4 %

BDE99: 20.5 µg/mL U=4 %

BDE153: 1.43 µg/mL U=4 %

N/A

VNIIM NIST 2257 see above N/A

KRISS AccuStandard

BDE 47, 99, and 153 were

98.75%, 95.50 %, and 95.83 %,

respectively

GC-FID assigned values

confirmed with SRM2257

TÜBITAK

UME

Chiron neat

substances

BDE47: 99.2±0.03 %

BDE99: 98.7±0.02 %

BDE153: 99.5±0.05 %

determined by qNMR

(SRM 1944 used for

confirmation, but BDEs

values are not certified)

GLHK NIST 2257

BDE47: 2.09±0.16 µg/g

BDE99: 2.127 ± 0.090 µg/g

BDE153: 2.048 ± 0.068 µg/g

N/A

NMIA Wellington Lab.

BDE47: 50 ± 2.5 µg/mL

BDE99: 50 ± 2.5 µg/mL

BDE153: 50 ± 2.5 µg/mL

10 % toluene in nonane

values assigned by

comparison to SRM 2257,

GBW(E)081124 and

GBW(E)081125

BAM NIST 2257 see above N/A

NIST NIST 2257 see above N/A

LNE AccuStandard

BDE47: U = 2 %

BDE99: U = 2 %

BDE153: U = 2.5 %

values assigned using

SRM 2257

NMIJ NIST 2257 see above N/A

CENAM CIL

BDE47: 49.8±2.5 µg/mL

BDE99: 50.0±2.5 µg/mL

BDE153: 50.0±2.5 µg/mL

values assigned against

SRM 2257

INMETRO Results not submitted


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6. ANALYTICAL METHODS EMPLOYED

The participating laboratories were encouraged to apply analytical methods of their own

choice. The methods for sample preparation, the analytical techniques for instrumental

analysis, the internal standards as well as the calibration type used in CCQM-K102 are

summarised in Table 6. The full details on the analytical methods, as reported by each

participant, are given in ANNEX A.

For the extraction step, most participants (9 out of 14) applied pressurised liquid

extraction (PLE), while 2 participants choose Soxhlet. NMIJ was the only one using

ultrasonic extraction. Solvents employed were hexane, acetone, CH2Cl2, toluene and

various mixtures thereof.

Clean-up of the sample was achieved mostly using solid phase extraction (SPE) and/or

multi-layer silica and Al2O3 columns.

Regarding the instrumental analysis, the splitting among different detection modes was

almost equal: 5 laboratories used GC-MS/MS, 5 laboratories used GC-MS (among which

NMISA using a TOF instrument), 4 laboratories used GC-HRMS. The ionisation mode

was electron impact for all laboratories, except JRC-IRMM which opted for negative

chemical ionisation.

All laboratories quantified with IDMS using the corresponding 13

C labelled PBDEs,

except JRC-IRMM which used internal standard quantification with BDE 77. Most

laboratories applied single-point calibration, while the rest choose for bracketing or 3 to 6

point calibration or a combination thereof.


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Table 6. Summary of analytical methodologies used in CCQM-K102

participant

sample

intake /

unit(s) #

(pre-treatment)

extraction clean-up

instrumental

technique internal standard(s) calibration

JRC-IRMM 2 g / 56 (Cu) ASE hexane/acetone

3:1 SPE, elution with hexane GC-MS (NCI) BDE 77

5 point internal

standard calibration

LGC 2 g / 48

(wetting overnight with

900 μL of ultrapure H2O)

Soxhlet, hexane/acetone

1:1

Step 1: SPE Acid silica/Florisil/Basic

silica/ Florisil

Step 2: LC Hypercarb fractionation

GC-MS 13

C BDEs 47, 99, 153

Bracketed Double

Exact Matching

IDMS

NMISA 1 g / 7, 50 ASE,

CH2Cl2: hexane 3:1

clean-up with Cu and neutral Al2O3

directly in the ASE cell GC-TOF MS

Wellington

Laboratories 13

C BDEs

solutions

IDMS followed by

bracketing double

IDMS

NIM 2 g / 17, 34,

67

ASE,

CH2Cl2: hexane 1:1

SPE combination of Alumina and

HLB, elution CH2Cl2 : hexane 2:1 GC-MS/MS

CIL 13

C BDEs

solutions IDMS single point

VNIIM 2.5 g / 35,

65, 19

ASE, hexane/acetone 9:1,

SoxTherm, toluene

silica multilayer column, Al2O3,

neutral Cu GC-MS

CIL EPA Method 1614 13

C BDEs solution EO-

5277

IDMS short range

KRISS 2 g / 45 PLE

CH2Cl2

SPE cartridge (silica gel), elution

with hexane GC-MS

CIL 13

C BDEs 47, 99,

153

IDMS Single-point

exact matching

double ID

TÜBITAK

UME 2 g / 41

(Cu/Na2SO4 3:1 + inert

dispersing agent)

PLE, n-hexane: acetone 7:3

multilayer column: deactiv. alumina,

deactiv. silica, acidified sílica,

elution CH2Cl2 : n-hexane 1:1

GC-MS/MS

Wellington

Laboratories 13

C BDEs

47, 99, 153

IDMS single-point


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Table 6 cont.

participant

sample

intake /

unit(s) #

(pre-treatment)

extraction clean-up

instrumental

technique internal standard(s) calibration

GLHK 2.5 g / 27,

52, 83 Soxhlet, CH2Cl2

Multi-layer silica gel and alumina

columns in sequence GC-HRMS

CIL 13

C BDEs

solutions

Exact matching

IDMS

NMIA 2 g / 21,

37, 53, 81

ASE, Toluene (other

conditions and Soxhlet used

for confirmation)

H2SO4 and H2O wash, automated SPE

with multi-layer silica (neutral, acidic,

basic) and Al2O3

GC-HRMS

Wellington

Laboratories 13

C

BDEs 47, 99, 153

Exact-matching

single-point double

IDMS with

bracketing

BAM 1.5 g / 15,

42 PLE

Al2O3 and multilayer silica (neutral,

acidic, basic) columns GC-MS

CIL 13

C BDEs 47, 99,

153

IDMS, 6 point

calibration

NIST 3 g / 57

(mix with granular

precombusted Na2SO4)

PLE, CH2Cl2

deactivated Al2O3 column, elution with

CH2Cl2:hexane 35:65 GC-MS/MS

13C BDEs 47, 99, 153

5 point calibration

curve followed by

bracketing

LNE 1 g / 8, 44,

77 ASE, CH2Cl2 pretreated silica and Al2O3 column GC-MS/MS

CIL 13

C BDEs 47, 99,

153

IDMS multipoint

calibration curve

NMIJ

14.8 –

16.3 g /

11, 39, 73

ultrasonic extraction CH2Cl2

Cu and Na2SO4; acidic silica column,

elution CH2Cl2/hexane 2/8; SPE,

elution acetone/hexane 1/9

GC-HRMS

CIL EPA Method

1614 13

C BDEs

solution EO-5277

IDMS, 3 point

calibration

CENAM 2 g / 16 Soxhlet, CH2Cl2 SPE silica GC-MS/MS CIL

13C BDEs 47, 99,

153 IDMS single-point

INMETRO Results not submitted


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6.1 Dry mass determination

All participants followed the prescribed protocol for dry mass determination (see section

4), entailing a drying oven procedure, except TÜBITAK UME which used Karl Fischer

titration and performed the analysis using a sample intake of 0.2 g (instead of 1 g).

Correction factors calculated by the participants and applied to report the results on a dry

mass basis corresponded to an amount of volatiles between 0.23 and 1.01 % (m/m), with

the large majority of laboratories finding a % m/m value between 0.4 and 0.8. The details

on the dry mass determinations are reported in ANNEX A.

7. RESULTS

The measurement results officially submitted for BDE 47, 99 and 153 in CCQM-K102

are reported in Table 7, 8 and 9, respectively.

LNE, NIM and NIST provided results also for BDE 209 (optional measurand of the

study). These results are presented in ANNEX B, Table 1-B.

Table 7. BDE 47 results in CCQM-K102

Participant

Mass

fraction

(µg/kg)

Combined

standard

uncertainty

u (µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Volatiles

content

(% m/m)

JRC-IRMM 16.3 0.70 2 1.4 0.65

LGC 15.81 0.41 2 0.82 0.69

NMISA 14.04 0.94 2.025 1.90 0.40

NIM 14.65 0.35 2 0.70 0.70

VNIIM 14.8 0.58 2 1.2 0.63

KRISS 16.63 0.23 2.45 0.56 0.63

TÜBITAK UME 16.91 0.76 2 1.52 0.59

GLHK 14.8 0.7 2 1.4 0.23

NMIA 16.2 1.0 2.12 2.1 0.50

BAM 14.364 0.9114 2 1.823 1.01

NIST 15.6 0.65 4.3 2.8 0.75

LNE 15.77 2.99 2 5.98 0.63

NMIJ 13.8 0.55 2 1.1 0.77

CENAM 16.43 0.91 2.18 1.98 0.48

INMETRO Result not submitted


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Participant

Mass

fraction

(µg/kg)

Combined

standard

uncertainty

u (µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Volatiles

content

(% m/m)

JRC-IRMM 34.1 3.33 2 6.7 0.65

LGC 33.69 0.61 2 1.22 0.69

NMISA 48.12 4.01 2.025 8.13 0.40

NIM 35.00 0.78 2 1.56 0.70

VNIIM 32.8 0.89 2 1.8 0.63

KRISS 36.0 1.1 2.78 3.1 0.63

TÜBITAK UME 39.50 2.04 2 4.07 0.59

GLHK 31.21 1.35 2 2.70 0.23

NMIA 34.4 2.3 2.03 4.7 0.50

BAM 31.299 1.174 2 2.349 1.01

NIST 41.1 0.85 4.3 3.7 0.75

LNE 30.93 3.83 2 7.66 0.63

NMIJ 31.2 0.77 2 1.55 0.77

CENAM 35.14 1.43 2.57 3.69 0.48


Upon discussion at the OAWG meeting in spring 2015, when the results were presented

for the first time, some of the laboratories deemed necessary a thorough review of their

submitted results, in relation to the influence of a proper chromatographic separation on

the measured mass fraction values, especially for BDE 153.

The further review of the results resulted in revised datasets, both for mass fraction

values and/or uncertainty values for some of the laboratories, which caused the

withdrawing of some results for BDE 99 and 153 from consideration in the calculation of

the KCRV. The data re-submitted by some laboratories are reported in ANNEX B,

Tables 2-B, 3-B and 4-B with the explanation substantiating the applied revision. Figures

2-B, 3-B and 4-B in ANNEX B report the graphs showing both officially submitted and

revised results with their standard uncertainties.

The calculations of the KCRV and DoE were carried out only taking into account valid

officially submitted results and uncertainties, see below Section 9.


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Participant

Mass

fraction

(µg/kg)

Combined

standard

uncertainty

u (µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Volatiles

content

(% m/m)

JRC-IRMM 5.9 0.74 2 1.5 0.65

LGC 6.38 0.21 2 0.42 0.69

NMISA 7.12 0.52 2.013 1.05 0.40

NIM 7.177 0.19 2 0.38 0.70

VNIIM 6.11 0.16 2 0.32 0.63

KRISS 5.78 0.20 2.57 0.51 0.63

TÜBITAK UME 11.03 0.72 2 1.45 0.59

GLHK 5.757 0.308 2 0.617 0.23

NMIA 6.24 0.36 2.02 0.73 0.50

BAM 7.489 0.2338 2 0.457 1.01

NIST 8.97 0.2 4.3 0.87 0.75

LNE 7.03 1.75 2 3.50 0.63

NMIJ 6.28 0.31 2 0.62 0.77

CENAM 7.72 0.86 2.20 1.88 0.48


8. APPROACHES TO UNCERTAINTY BUDGET ESTIMATION

The (main) contributions to the uncertainty budgets declared by the participating

laboratories are summarised in Table 10. The full details of the uncertainty evaluation

reported by the laboratories are given in ANNEX C.


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Table 10. Overview of participants' uncertainty budget estimation in CCQM-K102

Participant Sources of main uncertainty contributions

BAM

ratio native/internal standard (rsample)

intercept of calibration line (ic)

analyte remaining in the sample (corlod)

contribution from all weighings

extraction variability (Fex)

integration variability (Fch)

concentration of native standards (Fpurity)

CENAM

m0 Standard mass

mI0 Standard isotope mass

mx Sample mass

mIx Standard isotope mass in sample

R0 Response ratio of standards

Rx Response ratio of sample

w0 Mass fraction of standard

CFh Humidity correction factor

wx Mass fraction of measurand in sample

GLHK

The systematic uncertainty included:

uncertainty of calibration standard solution (ucal): preparation of

standard solution (weighing) and uncertainty of NIST BDE

standard solutions uncertainty due to preparation of sample blend and calibration blend

(uweighing)

the uncertainty due to moisture correction (umoisture)

The random uncertainty was calculated from the precision (standard

deviation) of multiple measurement results from three bottles.

10

)(

ilod

losspuritychex

samplealiquot

issolventislod

sample

sample

Fandcorwith

FFFFmm

cmmcor

sl

icrc

CFhwmRm

mRmw

Ix

xIx

x *0

00

0


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Table 10 cont.

JRC-IRMM

repeatability (n1=7)

intermediate precision (n2=1)

trueness (based on recovery)

preparation of calibration standards

22

2

2

1

2

* caltrue

iprepuu

n

RSD

n

RSDkU

Values inferred from method validation data

KRISS

Combination of systematic and random uncertainties as shown below:

Main contributions to the systematic term:

Uncertainty of gravimetric preparation for standard solutions

Uncertainty of gravimetric mixing for calibration isotope standard

mixtures

Area ratio native/labelled PBDE for the calibration mixture, observed by

GC/MS

LGC

LNE

Preparation of sample blends (weighings)

Calibration model

Preparation of calibration blend (weighings)

Precision

NIM Method precision

Calibration solution

n

suCu

22

systematics.p.,mean )(


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Table 10 cont.

NIST

measurement repeatability

water content determination

calibration (including uncertainty of the certified value of the calibration

solution)

The 95 % expanded uncertainties were estimated using the two-tailed Student’s t

value for two degrees of freedom, t(95,2) = 4.3

NMIA

Method Precision

Method Trueness

Standard

Moisture Content

Gravimetry

Isotope Amount Ratio

Blend Isotope Amount Ratio

NMIJ concentration of primary standard

repeatability from GC/MS analysis for sample

calibration curve

NMISA

Native solution added to calibration blend

Ratio of peaks areas of native/labelled in the samples

Measurement repeatability

Dry mass correction factor repeatability

TÜBITAK

UME

Bottom up approach considering the following sources:

Mass of sample intake

Labelled stock solution

Spiking of labelled stock solution

Recovery

Repeatability

Karl-Fisher water determination

Mass of final sample

VNIIM


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9. KEY COMPARISON REFERENCE VALUES CALCULATION

The OAWG has established criteria for the inclusion of results in the calculation of

the KCRV e.g., use of a properly validated method, calibration standards with a

metrologically traceable assigned purity value (CRM or material which purity has

been suitably assessed by the participant). The KCRVs established for the CCQM-

K102 will be the reference values for the CCQM-P138.

Discussion at the CCQM meeting in April 2015 in Sèvres pointed to the possibility

of chromatographic interferences for BDE153 leading to some results being biased

high. Several laboratories deemed it necessary to perform a follow-up study during

2015 to investigate this issue. As a result, several laboratories submitted revised data

and/or uncertainties and three of them decided to withdraw some of the officially

submitted results. The revised results were not considered for the calculation of the

KCRVs.

LNE and VNIIM reviewed uncertainty contributions to their measurement

uncertainty budgets and re-submitted lower and slightly higher estimates,

respectively. NMISA discovered a calculation error for the BDE 99 and 153 values

and withdrew these results.

NIST also withdrew the submitted results for BDE 99 and 153, after a follow-up

study using a chromatographic column of 30 m (instead of 10 m, see ANNEX B).

TÜBITAK UME withdrew the result for BDE 153, also as a consequence of the

employment of a longer column for analysis (60 m instead of 15 m). In both cases

the use of longer columns removed interferences that had biased the originally

reported results.

At the OAWG meeting in October 2015, D. Duewer (NIST) presented an analysis

of different KCRV estimators: mean, median and Der-Simonian Laird (weighted

based on uncertainties). As no significant difference existed among these estimators,

the final suggestion was to use a robust and simple estimator like the median that

was considered as fit-for-purpose for this dataset. The OAWG also made a thorough

examination of the data and discussed a possible inclusion of a heterogeneity factor

to enlarge the KCRV uncertainty due to concerns from some participants about the

potential effect of sample heterogeneity. Regarding the introduction of such a

heterogeneity factor to enlarge the KCRV uncertainty, it was deemed not necessary

in the view of 1) median and MADe are reasonable approximations to the

homogeneity values, 2) 8 out of 13 (for BDE 47) and 7 out of 11 (for BDE 99 and

153) laboratories submitted results based on 2 to 4 bottles and 3) the number of

participants was large enough to capture possible effects of heterogeneity (see also

[11]). Nevertheless, a vote on inclusion of a heterogeneity factor was carried out and

the majority voted against (two in favour out of eleven participants present).

Furthermore, the OAWG took the final decision to use the median as the estimate

for the KCRV.

In the final calculation of the KCRVs, the data from TÜBITAK UME were not

included due to the fact that the laboratory did not follow the prescribed method for

the dry mass determination, using Karl-Fischer instead of the oven-drying that was

provided in the study protocol. This creates a traceability issue, because the amount


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of volatiles determined with the oven-drying is method–dependant, while with the

Karl-Fischer method only water is determined.

The assessment of the traceability of the purity values used by the participants was

considered with regard to the results' inclusion in the KCRV calculation. KRISS

results were initially excluded from the KCRVs’ calculation because the purity of

the commercial calibrants employed was reported as only being assessed by GC-

FID. An in-depth discussion revealed that SRM2257 was used to confirm the purity

assigned by GC-FID thereby performing a transfer-value assignment similarly to

other laboratories (see Table 5). Their results were therefore included in the final

calculation of the KCRVs.

As a result of the overall above-mentioned evaluation, the KCRVs were calculated

as the median of thirteen valid results for BDE 47 and eleven results for BDE99 and

BDE 153, respectively (see Table 11a, b, c and Figures 3a, b, c).

The KCRV, its associated standard uncertainty and the results of the participants

(with their standard uncertainties) in CCQM-K102 are reported in Table 11a, b, c

and Figures 3a, b, c for BDE 47, 99 and 153, respectively. Graphs reporting the

KCRV, its expanded uncertainty and the laboratories' expanded uncertainties are

given in ANNEX D.

Table 11a. BDE 47 in CCQM-K102

KCRV and associated uncertainty (µg/kg dry mass basis)

KCRV (median) 15.60

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 1.19

𝑆𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦 (𝑢) = 1.25 ∗ 𝑀𝐴𝐷𝑒/√𝑛 (n=13) 0.41

U95 % (k*u) k = 2 0.82

Figure 3a. CCQM-K102: KCRV and its standard uncertainty for BDE 47.

Participants' results are also displayed with their standard uncertainties.


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Table 11b. BDE 99 in CCQM-K102

KCRV and associated uncertainty (µg/kg dry mass basis)

KCRV (median) 33.69

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 2.15


U95 % (k*u) k = 2 1.62

Figure 3b. CCQM-K102: KCRV and its standard uncertainty for BDE 99.



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Table 11c. BDE 153 in CCQM-K102

KCRV and associated uncertainty for (µg/kg dry mass basis)

KCRV (median) 6.28

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 0.74


U95 % (k*u) k = 2 0.56

Figure 3c. CCQM-K102: KCRV and its standard uncertainty for BDE 153.


10. DEGREES OF EQUIVALENCE (DOE) CALCULATION

The Degrees of Equivalence (DoE) of each result with the KCRV is expressed

quantitatively by two components: a value component and an uncertainty

component. The DoE and its uncertainty between an NMI result and the KCRV is

calculated within CCQM according to the following equations:

- the value component is di = xi - xref

Where, di is the degree of equivalence between the participant's result xi and the

KCRV = xref.

- the uncertainty component is U(di) = k * u(di)

Where, the expanded uncertainty U(di) is calculated by combining the standard

uncertainties ui of xi and uref of xref and multiplying by the coverage factor k=2 as

follows: U(di) = 2 ∗ √ui2 + uref

2 .


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Tables 12a, b and c list the absolute and relative [% di = 100·di/KCRV and U (% di)

= 100·U(di)/KCRV] DoE and their associated expanded uncertainty; the last column

shows the ABS (di)/U(di).

A reported result xi is consistent with the KCRV in CCQM-K102 within its stated

uncertainty at the 95 % confidence level when ABS (di)/U(di) < 1 (visually: when

the uncertainty bars in Figures 4a-e cross the red line).

Figures 4a-e display the absolute di ± U (di) and the relative % di ± U (% di) for

BDE 47, BDE 99 and BDE 153, respectively, in CCQM-K102.

Table 12a. BDE 47 in CCQM-K102

Degrees of equivalence [di] and expanded uncertainties [U(di)]

BDE 47

participant µg/kg %

di U(di) % di % U(di) di /U(di)

NMIJ -1.80 1.374 -11.5 8.8 1.31

NMISA -1.56 2.052 -10.0 13.2 0.76

BAM -1.24 2.000 -7.9 12.8 0.62

NIM -0.95 1.080 -6.1 6.9 0.88

GLHK -0.80 1.624 -5.1 10.4 0.49

VNIIM -0.80 1.422 -5.1 9.1 0.56

NIST 0.00 1.538 0.0 9.9 0.00

LNE 0.17 6.036 1.1 38.7 0.03

LGC 0.21 1.162 1.3 7.4 0.18

NMIA 0.60 2.163 3.8 13.9 0.28

JRC-IRMM 0.70 1.624 4.5 10.4 0.43

CENAM 0.83 1.997 5.3 12.8 0.42

KRISS 1.03 0.942 6.6 6.0 1.09

TÜBITAK UME 1.31 1.728 8.4 11.1 0.76

The entries in italic are results not included in the KCRV calculation


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Figure 4a. Absolute degrees of equivalence for BDE 47 in CCQM-K102.

The vertical bars correspond to ±U(di).

The horizontal red line marks the zero deviation from the KCRV.

Figure 4b. Relative degrees of equivalence for BDE 47 in CCQM-K102.

The vertical bars correspond to ±U(%di).


-6.0

-4.0

-2.0

0.0

2.0

4.0

6.0

De

gre

e o

f e

qu

ival

en

ce µ

g/kg

BDE 47

-40.0

-30.0

-20.0

-10.0

0.0

10.0

20.0

30.0

40.0

De

gre

e o

f e

qu

ival

en

ce %

BDE 47


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Table 12b. BDE 99 in CCQM-K102


BDE 99



LNE -2.76 7.830 -8.2 23.2 0.35

NMIJ -2.49 2.236 -7.4 6.6 1.11

GLHK -2.48 3.149 -7.4 9.3 0.79

BAM -2.39 2.853 -7.1 8.5 0.84

VNIIM -0.89 2.407 -2.6 7.1 0.37

LGC 0.00 2.029 0.0 6.0 0.00

JRC-IRMM 0.41 6.854 1.2 20.3 0.06

NMIA 0.71 4.877 2.1 14.5 0.15

NIM 1.31 2.250 3.9 6.7 0.58

CENAM 1.45 3.287 4.3 9.8 0.44

KRISS 2.31 2.733 6.9 8.1 0.85

TÜBITAK UME 5.81 4.390 17.2 13.0 1.32

NIST 7.41 2.349 22.0 7.0 3.15

NMISA 14.43 8.182 42.8 24.3 1.76



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Figure 4c. Absolute degrees of equivalence for BDE 99 in CCQM-K102.



Figure 4d. Relative degrees of equivalence for BDE 99 in CCQM-K102.



-10.0

-6.0

-2.0

2.0

6.0

10.0

14.0

18.0

De

gre

e o

f e

qu

ival

en

ce µ

g/kg

BDE 99

-40.0

-20.0

0.0

20.0

40.0

60.0

80.0

De

gre

e o

f e

qu

ival

en

ce %

BDE 99


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Table 12c. BDE 153 in CCQM-K102


BDE 153



GLHK -0.52 0.832 -8.3 13.2 0.63

KRISS -0.50 0.687 -8.0 10.9 0.73

JRC-IRMM -0.38 1.582 -6.1 25.2 0.24

VNIIM -0.17 0.644 -2.7 10.3 0.26

NMIA -0.04 0.911 -0.6 14.5 0.04

NMIJ 0.00 0.835 0.0 13.3 0.00

LGC 0.10 0.699 1.6 11.1 0.14

LNE 0.75 3.544 11.9 56.4 0.21

NMISA 0.84 1.181 13.4 18.8 0.71

NIM 0.90 0.676 14.3 10.8 1.33

BAM 1.21 0.729 19.3 11.6 1.66

CENAM 1.44 1.809 22.9 28.8 0.80

NIST 2.69 0.687 42.8 10.9 3.91

TÜBITAK UME 4.75 1.545 75.6 24.6 3.08



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Figure 4d. Absolute degrees of equivalence for BDE 153 in CCQM-K102.



Figure 4e. Relative degrees of equivalence for BDE 153 in CCQM-K102.



11. CORE COMPETENCIES AND HOW FAR DOES THE LIGHT SHINE?

The participation to the Track A "Low polarity analytes in abiotic matrix" CCQM-

K102 study, polybrominated diphenyl ethers in sediment, was intended to

demonstrate the capability of NMIs/DIs of analysing non-polar organic molecules in

-3.0

-2.0

-1.0

0.0

1.0

2.0

3.0

4.0

5.0

6.0

De

gre

e o

f e

qu

ival

en

ce µ

g/kg

BDE 153

-40.0

-20.0

0.0

20.0

40.0

60.0

80.0

100.0

De

gre

e o

f e

qu

ival

en

ce %

BDE 153


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abiotic dried matrices using procedures requiring extraction from the matrix, clean-

up from interfering substances, analytical separation, selective detection and final

quantification. The details of the specific approaches/techniques used by each

participant underpinning their competencies are reported in the Core Competency

Tables given in ANNEX E.

More specifically, the comparison was designed to demonstrate the laboratory's

competence in the quantification of organic molecules in the approximate range of

molecular weights from 100 to 800 g/mol, having polarity corresponding to pKow <

-2 and for the range of mass fraction 1-1000 μg/kg in abiotic dried matrices.

12. CONCLUSIONS

Most of the participants to CCQM-K102 were able to demonstrate or confirm their

capabilities in the analysis of non-polar organic molecules in abiotic dried matrices.

Throughout the study, it became clear that matrix interferences can influence the

accurate quantification of the PBDEs, if the analytical methodology applied is not

appropriately adapted and optimised. This comparison shows that quantification of

PBDEs at the µg/kg low-middle range in a challenging environmental abiotic dried

matrix can be achieved with relative expanded uncertainties below 15 % (more than 70

% of participating laboratories), well in line with the best measurement

performances in the environmental analysis field.

13. ACKNOWLEDGEMENTS

The study coordinator wishes to thank the JRC-Geel Reference Materials Processing

Team and Giovani Kerckhove of the Engineering Materials Laboratory for the

electron microscopy work in the assessment of the study material homogeneity. A

particular thank goes to D. Duewer for precious statistical and pragmatic advice and

to L. Mackay, chair of the OAWG, for her advice.

14. REFERENCES

[1] J. Xu, Z. Gao, Q. Xian, H. Yu, J. Feng (2009) Environ. Pollut. 157: 1911-16.

[2] M. Osako, Y.J. Kim, S. Sakai (2004) Chemosphere. 57: 1571-79.

[3] A.Hassanin et al. (2004) Environ. Sci. Technol. 38:738-745.

[4] J. de Boer, P.G. Wester, A. van der Horst, P.E.G. Leonards (2003) Environ.

Pollut. 122: 63-74.

[5] R.A. Hites (2004) Environ Sci Technol. 38(4): 945–956.

[6] R.J. Law et al. (2008) Chemosphere 73(2): 223–241.

[7] (a) Directive 2003/11/EC (24th

Amendment to Directive 76/769/EEC), (b)

Directive 2002/95/EC.

[8] Scientific opinion on PBDEs in food, EFSA Panel on contaminants in the Food

chain (CONTAM), EFSA Journal (2011) 9(5): 2156.

[9] Braekevelt, E., Tittlemier, S.A., Tomy, G.T. (2003) Chemosphere 51: 563–567.

[10] T.P.J Linsinger, J. Pauwels, A.M.H. van der Veen, H. Schimmel, A. Lamberty

(2001) Accred. Qual. Assur. 6: 20-25.

[11] CCQM Guidance note: Estimation of consensus KCRV and associated Degrees

of Equivalence" version 10, 2013-04-12.


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ANNEX A

Full details of the analytical methods employed by the participants in CCQM-

K102

BAM

Information about the analytical procedure (N.B. especially for details not mentioned in the Core Competency Table)

Sample intake used for analysis g

Sample pre-treatment (if applicable)

Extraction method/conditions PLE

T=100 °C

p=140 bar

static-time: 10 min

cycles 3

Clean-up procedure Al2O3 column chromatography (1 cm of sodium sulfate; 25 g of

(e.g., SPE, GPC) alumia oxide; 1 cm of sodium sulfate)

Multilayer column chromatography (1,0 g of sodium sulfat / 0,5 g silica

gel/silver nitrate; 0,2 g of silica gel; 1,0 g of silica gel/sulfuric acid; 0,2 g

of silica gel; 0,5 g of silica gel/sodium hydroxid

Analytical instrumentation used GC-MS (Agilent 7890A with 5975C MSD)

(e.g., GC-MS ) Cold injektion system (CIS): 90°C for 0.01 min then 10°C/sec to 360°CGC settings for 15 min; injection volume 2µl splitless

(e.g., injection mode, T and volume; carrier gas ZB-5 HT Inferno 15 m x 250 µm x 0.1 µm; flow 1 ml/min (He)

and flow; column type, oven temperature program) Oven programm: 80 °C for 1 min then 20 °C/min to 340 °C for 5 min

MS settingsMS settings EI-mode; gain factor 10 (Em voltage 2388) source 300°C / quad 150 °C

(e.g., MS mode, monitored ions, T, electron Ions for BDE47/MBDE47: 325.9; 337.9; 483.7; 485.7; 495.8; 497.8; 499.8

multiplier voltage, gas) Ions for BDE99/MBDE99: 405.7; 415.8; 417.8; 563.6; 565.6; 575.7; 577.7

Ions for BDE153/MBDE153: 485.7; 495.8; 497.8; 641.5; 643.5; 655.6; 657.7

Water content determination(please describe the procedure, if deviating intake: 1,2 - 1,4 g

from the one prescribed in the protocol drying temperature: 105 °C

and the calculation of dry mass correction factor)

Calibration type / details IDMS

(e.g., single-point, bracketing / BDE47: 18.8; 37.6; 97.3; 196.1; 301.2; 362.8 ng/ml

external calibration, internal standard calibration, IDMS) BDE99: 19.1; 38.3; 99.0; 199.6; 306.5; 369.2 ng/ml

BDE153: 18.4; 36.9; 95.3; 192.2; 295.1; 355.2 ng/ml

Calibration standards NIST 2257

(e.g., source, purity, uncertainty) BDE 47: 2.09 ± 0.16 µg/g

BDE 99: 2.127 ± 0.090 µg/g

BDE 153: 2.048 ± 0.068 µg/g

Internal standards used 13C12, 99% BDE-47 (CIL) about 239 ng/ml(Please specify the compounds, and at which stage 13C12, 99% BDE-99 (CIL) about 497 ng/ml

13C12, 99% BDE-153 (CIL) about 110 ng/ml

1.5

(e.g., PLE, Soxhlet / T, P, duration and cycles, solvents)


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CENAM



Sample pre-treatment (if applicable) none

Extraction method/conditions Automated Soxhlet Extraction (Soxhlet Standard)

Solvent used dichloromethane

Lower heating level 7

Time 4 h

Clean-up procedure SPE Silica

(e.g., SPE, GPC)

Analytical instrumentation used GC-MS/MS Injection volume 2 µL

(e.g., GC-MS ) Injection mode: Splitless MSD transfer line:300 °CGC settings Injector temperature: 280 °C Carrier gas: Helium

(e.g., injection mode, T and volume; carrier gas Column: HP-5MS 15m x 250 µm x 0.25µm Flow:0.9 mL/min

and flow; column type, oven temperature program) Oven: 60 °C (1 min); 40°C/min. to 170 °C (0 min); 10 °C/min. to 310 °C(3 min)

MS mode: MRM Source temperature: 230 °C Quad MS1 temp: 200 °CMS settings Monitored ions EMV: 1194.9; 1263.9 and 1306.1 CID Gas: Helium

(e.g., MS mode, monitored ions, T, electron 485.7 - 325.7 325.8-218.8 497.7 - 337.7 337.8 - 228.8

multiplier voltage, gas) 565.6 - 405.6 563.6 - 403.7 577.6 - 417.6 575.6 - 415.7

483.7 - 323.6 643.6 - 483.6 655.6 - 495.6 495.7 - 335.6

Water content determination Protocol method: Two 1 g subsamples were measured from bottle 0016

(please describe the procedure, if deviating by oven method described in the protocol.

from the one prescribed in the protocol


Dry mass correction factor: CF = 100 + % humidity / 100


(e.g., single-point, bracketing / Single point, verified with 3 independent preparations

external calibration, internal standard calibration, IDMS)

Calibration standards Source (µg/mL) U (µg/mL)

(e.g., source, purity, uncertainty) BDE-47 CIL 49.8 2.5

BDE-99 CIL 50.0 2.5

BDE-153 CIL 50.0 2.5

Internal standards used BDE-47 (13C12) CIL 50 1.3

(Please specify the compounds, and at which stage BDE-99 (13C12) CIL 50.2 4.2

BDE-153 (13C12) CIL 47.2 2.3

Purity assessment of the calibrant (if applicable)

(e.g. methods used for value assignment/verification) The uncertainty of the calibration solutions were estimated at CENAM

since the pure calibrants were not possible to procurement them.

The calibration solutions were verified using SRM2257

2



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GLHK

Informat ion about the analy t ical procedure (N.B. especially for details not mentioned in the Core Competency Table)

Sample intake used for analys is g

Sample pre-t reatment (i f applicable) NA

Extract ion method/condit ions Soxhlet extraction with dichloromethane for 16 hours

Clean-up procedure Clean-up in sequence by:-

(e.g., SPE, GPC) 1) Multi-layer silica gel column

Pack a glass chromatographic column from bottom to top with activated silica gel,

basic silica gel, acidic silica gel, activated silica gel, anhydrous Na2SO4

2) Alumina column

Analy t ical ins t rumentat ion used HRGC-HRMS

(e.g., GC-MS ) Splitless Injection at 290 °C; UHP He carrier gas at 1.2 ml/min (Constant flow);

GC settings Separation by Agilent DB-XLB column (30m x 0.25mm, 0.1µm)

(e.g., injection mode, T and volume; carrier gas Injection volume 3uL;

and flow; column type, oven temperature program) Oven program:100°C (hold for 3 min), 5°C/min ramped to 320 °C (hold for 5 min)

MS settings SIR mode:

(e.g., MS mode, monitored ions, T, electron BDE-99 m/z 563.6216 and 565.6196; 13C-BDE-99 m/z 575.6619 and 577.6598

multiplier voltage, gas) BDE-153 m/z 641.5322 and 643.5302 ; 13C-BDE-153 m/z 655.5704 and 657.5683

BDE-47 m/z 485.7111 and 483.7132 ; 13C-BDE-47 m/z 497.7514 and 499.7493

Water content determinat ion NA

(please describe the procedure, if deviating



Calibrat ion type / details Exact matching IDMS

(e.g., single-point, bracketing /


Calibrat ion s tandards NIST SRM 2257

(e.g., source, purity, uncertainty) BDE-47 2.09 ± 0.16 µg/g

BDE-99 2.127 ± 0.090 µg/g

BDE-153 2.048 ± 0.068 µg/g

Internal s tandards used Standard solutions from Cambridge Isotope Laboratories Inc.

(Please specify the compounds, and at which stage were 13C12-BDE-47, 50 µg/mL;13C12-BDE-99, 50 µg/mL;13C12-BDE-153, 50 µg/mL

2.5



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Sample pre-treatment (if applicable) N/A

Extraction method/conditions ASE

Pressure: 1500 psi; Temperature: 120 °C; Preheat time: 5 min;

Static time: 6 min; Flush volume: 150 %; Purge time: 120 sec;

Static cycles: 3; Solvents: hexane/acetone 3:1

Clean-up procedure SPE

(e.g., SPE, GPC) Conditioning: 3 mL; Load: 1 mL sample; elution: 4 mL;

Flow speed: 1 mL/min; Cartridge: Bond-Elute PCB (1g, 3 mL)

Solvent: Hexane

Analytical instrumentation used GC-MS

(e.g., GC-MS )GC settings Pulsed splitless; Initial temp: 90 °C; Pressure: 2.69 psi; Carrier gas: He

(e.g., injection mode, T and volume; carrier gas Total flow: 103.3 mL/min; column: DB-5MS, 15 m * 0.25 mm * 0.25 µm

and flow; column type, oven temperature program) T Progr.: 90 °C (1.5 min), 20 °C/min to 270 °C (0 min), 10 °C/min to 285 °C

(0 min), 35 °C/min to 300 °C (7 min), 50 °C/min to 320 °C (5 min)

MS settings NCI; monitored ions: 79, 81; AUX 250 °C, Methane, 25 eV

(e.g., MS mode, monitored ions, T, electron

multiplier voltage, gas)

Water content determination According to the CCQM-K102/P138 protocol




Calibration type / details internal standard 5 points calibration



Calibration standards NIST SRM 2257

(e.g., source, purity, uncertainty) Certified concentrations of compounds:

PBDE 47 - 2.09 ± 0.16 ug/g

PBDE 99 - 2.127 ± 0.090 ug/g

PBDE 153 - 2.048 ± 0.068 ug/g

Internal standards used PBDE 77 (Accustandard), Purity (GC/MS) - 100%±5% (U )

(Please specify the compounds, and at which stage

were added) Added after weighing of the sample



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Extraction method/conditions Pressurized Liquid Extraction (PLE) method was used for sample extraction.

Extraction conditions were with temperature 100 °C , pressure 1500 psi, extraction time 5 min,

and number of extraction cycle 2 with dichoro methane as a solvent.

Clean-up procedure

(e.g., SPE, GPC) Cleaned up by using SPE cartridge (silica gel) with hexane as eluant

Analytical instrumentation used GC/MS (JMS-800D, Jeol)

(e.g., GC-MS ) GC condition

GC settings Injection mode: on-column injection (confirmatory: splitless injection)

(e.g., injection mode, T and volume; carrier gas Injection volume: 1 uL of sample

and flow; column type, oven temperature program) Carrier gas : He, 1.5 mL/min

MS condition

MS settings Ion source: EI mode

(e.g., MS mode, monitored ions, T, electron Resolution : 1000 (with confirmatory check by high resolution measurement at 10,000)

multiplier voltage, gas) primary monitoring channel: m/z 485.7 for BDE-47, m/z 403.8 for BDE-99, and m/z 483.7 for BDE-153

confirmatory channel: m/z 325.9 for BDE-47, m/z 563.6 for BDE-99, and m/z 643.5 for BDE-153

Water content determination We took more than 1 g of three subsamples from the bottle(No. 45),

(please describe the procedure, if deviating and then dried in the oven at 105 °C until constant mass is attained. It took 1~2 hrs.

from the one prescribed in the protocol The mean water content of each subsamples was 0.63%

and the calculation of dry mass correction factor) Dry mass correction factor is calculated as follows

X= water contents

Calibration type / details

(e.g., single-point, bracketing / Single-point with exact matching double ID for IDMS


Calibration standards Neat commercial calibrants for BDE-47, 99, 153 were from Accustandard.

(e.g., source, purity, uncertainty)

Internal standards used


Purity assessment of the calibrant (if applicable) For the determination of the purities of the primary reference materials of BDE-47, BDE-99, and BDE-153,

(e.g. methods used for value assignment/verification) Purities of the materials were assessed only with GC/FID analysis for structurally related impurities.

As the amounts of BDE-47, 99, and153 primary reference materials were limited

Karl-Fischer Coulometry for water content, thermogravimetric analysis for non-volatile inorganic residues,

and headspace-GC/MS for residual solvents could not be carried out.

The purities assessed by KRISS was indirectly confirmed by comparing the standard solution prepared

with the materials to NIST SRM 2257

Estimation of impurities (if applicable)

(e.g. type of impurity, mass fraction, uncertainty) GC/FID result Content unc DOF

BDE47 98.75% 0.002% 2

BDE99 95.50% 0.105% 2

BDE153 95.83% 0.006% 2

2


13C12-BDE-47, 13C12-BDE-99, and 13C12-BDE-153, purchased from CIL, were used

as the internal standards of BDE-47, BDE-99, and BDE-153, respectively.

The purities were assessed by KRISS with GC-FID, but water content and non-volatile impurities were

not tested due to the limited amounts of the materials. Purities: BDE 47, 99, and 153 were 98.75%,

95.50%, and 95.83%, respectively. The purities assessed by KRISS was indirectly confirmed by comparing

the standard solution prepared with the materials to NIST SRM 2257

)1/(1 xf


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Sample pre-treatment (if applicable) Wetting overnight adding 900 μL of ultrapure water

Extraction method/conditions Soxhlet extraction

Extraction time 24 hours

Extraction Solvent200 mL Hexane/Acetone 1:1

Clean-up procedure Step 1: SPE Acid silica/Florisil/Basic silica/ Florisil

(e.g., SPE, GPC) Step 2: LC Hypercarb fractionation

Analytical instrumentation used GC-EI-MS, equipped with Dean switch used as divert valve, single quadrupole Agilent 7890GC and 5975C MS

(e.g., GC-MS ) Splitless mode, Splitless time= 2 minGC settings Injector temperature 250 °C, 1 μL injection volume, He carrier gas F=1 mL/min

(e.g., injection mode, T and volume; carrier gas Colum DB-XLB 30 m x 0.25 mm x0.25 μm

and flow; column type, oven temperature program) Oven program: 90C held for 2 min; 40C/min to 290C, held for 0.5; 3C/min to 330C,

held for 5 min; 40C/min to 340C, held for 14 min MS settings SIM mode. BDE 47 Quan Ions: 485.6 Labelled 497.6, Qual ions 325.8, 337.8, 563.5

(e.g., MS mode, monitored ions, T, electron BDE 99 Quan Ions: 563.5 Labelled 575.6, Qual ions 403.7, 415.7, 643.4

multiplier voltage, gas) BDE 153 Quan Ions: 481.6/483.6/485.6 Labelled 493.6, 495.6, 497.6, Qual ions 643.4, 655.4, 721.0

Source 300C, Quadrupole 150C, EMV 1765 Gain 15.0

Water content determination As protocol, 2 aliquots, average value used for dry mass correction




Calibration type / details Bracketed Double Exact Matching IDMS



Calibration standards


Internal standards used BDE 47 13C

(Please specify the compounds, and at which stage BDE 99 13C

BDE 153 13C

Purity assessment of the calibrant (if applicable) Stock Concentration and Purity assessed by qNMR

(e.g. methods used for value assignment/verification)

Estimation of impurities (if applicable) N/A

(e.g. type of impurity, mass fraction, uncertainty)

2


All IS spiked at beginning of soxhlet extraction to enable extraction/equilibration

with the samples. Degredation of IS checked and not found to be an issue

Chiron neat materials >98% pure, 47 and 99

solids checked by qNMR, made into solutions to

be value assigned by qNMR for

concentration/purity


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Sample pre-treatment (if applicable) rehomogeneisation by shaking the flask about 2min

Extraction method/conditions extraction by ASE200 : 2 static phases of 6min , 100°C, 140 bars ,

100 % DCM

Clean-up procedure purification on a column of pretreated silica and alumina

(e.g., SPE, GPC)

Analytical instrumentation used GC/MS²

(e.g., GC-MS ) injection: 10 µl injected by solvent ventGC settings carrier gas: helium 1.2ml/mn

(e.g., injection mode, T and volume; carrier gas column: non polar type 5MS , length: 15m, di:0.25 mm, film:0.1 µm

and flow; column type, oven temperature program) temperature program: 50°C during 3.30 min , then 20°C/min until 150°C

during 0min, then 10°C/min until 260°C during 0 min then 40°C/min MS settings until 320°C during 7min.

(e.g., MS mode, monitored ions, T, electron electronic impact with 70eV, source temperature:180°C, transfert line

multiplier voltage, gas) température:290°C, MS/MS mode,

Water content determination drying 1g of sediment at 105°C +/- 2°C until 2 subsequent weightings

(please describe the procedure, if deviating do not differ more than 1 mg

from the one prescribed in the protocol dry mass correction factor= final mass/ initial mass of sediment


Calibration type / details IDMS with calibration curve



Calibration standards certified solutions from AccuStandard with 2% combined standard

(e.g., source, purity, uncertainty) uncertainty for PBDE 47 and 99 and 2.5 % for PBDE 153

Internal standards used C13 labeled compounds from Cambridge Isotope Laboratories


Purity assessment of the calibrant (if applicable) verified with SRM2257 from NIST


1



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Extraction method/conditions

Clean-up procedure

(e.g., SPE, GPC)

Analytical instrumentation used

(e.g., GC-MS )GC settings

(e.g., injection mode, T and volume; carrier gas

and flow; column type, oven temperature program)

MS/MS:

MS settings



Water content determination




Calibration type / details 1) Calibration method: Single point.







were added)

Purity assessment of the calibrant (if applicable)


Estimation of impurities (if applicable) No.

(e.g. type of impurity, mass fraction, uncertainty)

2


2) quantification mode:BDE-47,99,153: GC-IDMS, BDE-

209:internal standard calibration.

GBW(E)081124 , GBW08709 are all from NIM,China.

Each bottle were shaken by turning upside down for 30 minutes

before analysis

ASE / 100°C, 1500 psi, 4 cycles, dichloromethane : hexane =1:1 (v : v)

SPE/combination of Alumina and HLB，preconditioned with 15 mL

hexane, eluted with 3 * 3 mL mixed solvents (dichloromethane:hexane

=2:1)

GC-MS/MS

GC: splitless injection, 300 °C， 2 μL；He at 1.5 mL/min; DB-5 15 m

* 0.25 mm * 0.10 μm, 90 °C increased to 300 °C at 20 °C/min

Their uncertainty are 4% ,1.4% respectively (K=2 ).

The solution of BDE-47,99,153 (13C labled) and internal

standard(BDE-207) is from Cambridge Isotope Laboratories.

They are added to sample before extraction.

The purity of neat BDE-47,99,153 is determined by HPLC-DAD

and qNMR. The purity of BDE- 209 is determined by HPLC-DAD

and HPLC-ICP-MS.

MRM, monitored ions were summarized in table 1 at the bottom of

this part, ion source tempeature: 280 °C, gas: He;

2 separate portions of 1.0 g sediment from bottle No.0017 and 0034

were drying in an oven at (105 ± 2) °C until constant mass is attained

(for 4 hours).


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Table 1 Monitored ions in MRM mode

Compound Precusor Product

47 485.7 325.8

47 483.7 325.8

47 325.8 244.9

47L 337.6 256.9

47L 337.6 228

99 405.7 296.9

99 403.7 296.9

99L 417.7 307.8

99L 417.7 257.8

153 643.5 483.6

153 483.6 376.7

153 483.6 323.8

153L 655.5 495.6

153L 495.6 335.8


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Sample intake used for analysis 1 g for 209 determination


Extraction method/conditions Samples were mixed with granular precombusted sodium sulfate (650oC for 18 hours) and then transferred to a pressurized fluid extract. cell.

Internal standard was added and then samples were extracted with

dichloromethane (100 oC, equilibration 5 min, static time 5 min, cell

pressure 13.8 MPa, with three cycles using 1/3 of the solvent each time

Clean-up procedure Samples were cleaned up using 1.8 g deactivated alumina columns

(e.g., SPE, GPC) (5 % water mass fraction) by adding sample in hexane and eluting with

9 mL 35:65 dichloromethane:hexane (mass fraction)

Analytical instrumentation used GC-MS/MS; on column injection (2 uL); 60 oC initial temperature and then

(e.g., GC-MS ) in oven track mode. Carrier gas was helium at 1.4 mL/min for 10 min thenGC settings 3 mL for min. The column was a 10 m x 0.18 mm x 0.18 um film thickness

(e.g., injection mode, T and volume; carrier gas thickness 5 % methyl polysiloxane column coupled to a 1 m retention gap.

and flow; column type, oven temperature program) Oven program: 70 oC for 1 min, 40 oC/min to 175 oC and then 10 oC to 305 oC

(10 min hold) followed by 40 oC to 325 oC. Data acquired in MRM mode.MS settings Electron ionization was used (70 eV).



Water content determination 2 g to 3 g of the samples (n-5) were placed in glass culture tubes, dried

(please describe the procedure, if deviating 24 hours at 120 oC then cooled in a desiccator for 1 hour and re-weighed.



Calibration type / details Five point calibrat. curve that bracketed the expected sample response.

(e.g., single-point, bracketing / This was achieved for all compounds except BDE 209 in the K103/P138

external calibration, internal standard calibration, IDMS) which was 30% higher than the highest calibration point.

Carbon labeled PBDE 47, 99, 153 and 209 were used as internal

standards.

Calibration standards Calibration standards were prepared from SRMs 2257 and 2258.


Internal standards used Internal standards added before extraction and included(Please specify the compounds, and at which stage Carbon labeled PBDEs 47, 99, 153, and 209.

3 g for BDE 47, 99, and 153



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Sample pre-treatment (if applicable) Sediment sub-samples were spiked with solutions of carbon 13-labelled BDE47, 99 and 153

and then mixed by rotation overnight (at least 12 hours) prior to solvent extraction.

Extraction method/conditions Method 1: Accelerated Solvent Extraction (ASE) with 100% toluene: samples were extracted three times.

Extraction conditions: 2 static cycles of 8 minutes each at 150 oC and 1700 psi

Method 2: Hot Soxhlet Extraction with 100% toluene; 10 hours cycle time with constant reflux (for confirmation)

Method 3: ASE with ethanol/toluene (68:32) and then 100% toluene; samples were extracted

three times with each solvent (for confirmation)

Method 4: ASE with acetone/hexane (50:50): samples were extracted three times (for confirmation)

Clean-up procedure 1. Concentrated sulphuric acid wash (3 x 20 mL)

(e.g., SPE, GPC) 2. Ultra-high purity water wash (3 x 20 mL)

3. Gel permeation chromatography using dichloromethane (this clean-up step was found to be unnecessary for

the CCQM study sample; however, GPC clean-up was used for samples of NIST SRM1944, a reference material

used in this study)

4. Automated solid phase extraction with multi-layer silica (neutral, acidic, basic) and alumina

columns

Analytical instrumentation used 1. Gas chromatography with high resolution mass spectrometry (GC-HRMS) (Thermo Scientific DFS)

(e.g., GC-MS ) GC SettingsGC settings Injection mode: PTV Large Volume at 140 oC, injection volume: 1 uL

(e.g., injection mode, T and volume; carrier gas Carrier gas: Helium

and flow; column type, oven temperature program) Column: Agilent DB-5 (0.1 mm x 10 m, 0.1 um film thickness)

Carrier flow rate: 0.4 mL/minMS settings Temperature program: 80 oC hold for 6 minutes, ramp at 20 oC/min to 245 oC, ramp at 5 oC/min

(e.g., MS mode, monitored ions, T, electron to 290 oC, ramp at 10 oC/min to 325 oC, hold for 5 minutes


MS Settings

MS mode: EI, 45 eV, resolution ~10,000 (5% valley definition)

Source temperature: 285 oC

Electron multiplier voltage: 1520 V

Reference gas: PFK

Monitored ions: BDE 47 & 13C12-BDE 47 (483.71262, 485.71057, 487.70853, 497.75080, 499.74880)

BDE 99 & 13C12-BDE 99 (563.62109, 565.61904, 575.66130, 577.65930)

BDE 153 & 13C12-BDE 153 (481.69697, 483.69492, 493.73723, 495.73518)

2. Gas chromatography tandem mass spectrometry (GC-MS/MS) (Agilent 7000B GC-QQQ)

GC Settings

Injection mode: PTV Solvent Vent at 140 oC, injection volume: 5 uL

Carrier gas: Helium

Column: Phenomenex Zebron ZB-SemiVolatiles (0.18 mm x 10 m, 0.18 um film thickness)

Carrier flow rate: 0.4 mL/min

Temperature program: 80 oC hold for 4 minutes, ramp at 20 oC/min to 245 oC, ramp at 5 oC/min

to 290 oC, ramp at 10 oC/min to 325 oC, hold for 10 minutes

MS Settings

MS mode: EI, 50 eV

Source temperature: 300 oC

Electron multiplier voltage: 1639 V

Reference gas: FC-43

Monitored SRMs:

BDE 47: 487.7 > 327.9, 485.7 > 325.9, 483.7 > 325.9

13C12-BDE 47: 499.8 > 339.9, 497.8 > 337.9, 495.8 > 335.9

BDE 99: 565.6 > 405.8, 565.6 > 403.8, 563.6 > 403.8, 561.6 > 401.8

13C12-BDE 99: 577.7 > 417.8, 577.7 > 415.8, 575.7 > 415.8, 573.7 > 415.8

BDE 153: 643.5 > 483.6, 485.6 > 325.9, 483.6 > 323.9, 481.6 > 323.9

13C12-BDE 153: 657.6 > 497.7, 655.6 > 495.7, 653.6 > 493.7, 495.8 > 335.9

Water content determination Procedure: oven drying at 105 oC until constant mass achieved (<0.5 mg difference between

(please describe the procedure, if deviating consecutive readings) as per study protocol, sample size ~1 g

from the one prescribed in the protocol Dry mass correction factor: The dry mass correction factor was calculated as (1/(1-MC)) where MC is the average

and the calculation of dry mass correction factor) measured moisture content of the study sample.

The average measured moisture content in 6 sub-samples of the study material were 0.50%, 0.52%, 0.51%,

0.51%, 0.48% and 0.48%.

The dry mass correction factor determined was: 1.00502 with a standard u of 0.00093 (degrees of freedom = 5).

2



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NMIA cont.

Calibration type / details Exact-matching single-point double IDMS with bracketing



Calibration standards 1. Wellington Laboratories

(e.g., source, purity, uncertainty) BDE47: 50 ± 2.5 ug/mL in toluene (10%)/nonane

BDE99: 50 ± 2.5 ug/mL in toluene (10%)/nonane

BDE153: 50 ± 2.5 ug/mL in toluene (10%)/nonane

2. NIST SRM 2257

PBDE Congeners in 2,2,4-Trimethylpentane

BDE47: 2.09 ± 0.16 ug/g

BDE99: 2.127 ± 0.090 ug/g

BDE153: 2.048 ± 0.068 ug/g

3. NIM China Certified Reference Material GBW (E) 081124

Calibration Solution of Industrial Penta-BDE in Iso-octane

BDE47: 20.0 ug/mL (expanded relative uncertainty 6%, k = 2)



4. NIM China Certified Reference Material GBW (E) 081125

Calibration Solution of Industrial Octa-BDE in Iso-octane


Internal standards used Wellington Laboratories; Carbon 13-labelled BDE 47, 99 and 153 (13C12-BDE47,


were added)13C12-BDE99, 13C12-BDE153).

Sediment sub-samples were spiked with solutions of carbon 13-labelled BDE47, 99 and 153

and then mixed by rotation overnight (at least 12 hours) prior to solvent extraction.

Purity assessment of the calibrant (if applicable) Not applicable.

(e.g. methods used for value assignment/verification) Calibration standards prepared from the materials purchased from Wellington Laboratories

were assigned values by performing a comparison of standards with the NIST SRM2257 and

NIM China Penta and Octa Mix solutions.


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Extraction method/conditions The sediment was weighed and EO-5277 (CIL: 13C-labeled BDEs) 0.21 g-0.23 g

was spiked CH2Cl2 (30 mL) was added and extracted by ultrasonic (10 min)

The mixture was centrifuged to recover the supernatant (which was kept in

a brown bottle) CH2Cl2 (30 mL) was added to the precipitate, and extracted

again by ultrasonic (10 min). The mixture was centrifuged to recover the

supernatant (combine the supernatant in the brown bottle)

Clean-up procedure Solvent of the supernatant was replaced with hexane and concentr. to 1 mL

(e.g., SPE, GPC) Cu powder was added and shaken for 3 min to remove elemental sulfur

Anhydrous Na2SO4 was added and shaken for 3 min to remove moisture

The solution was passed through to a column packed with 3 g of silica

impregnated with concentrated sulfuric acid (44 %), then eluted with 25 mL

of CH2Cl2/hexane (20/80, v/v)

Solvent of the eluate was replaced with hexane and concentrated to 1 mL

The solution was loaded onto a SPE column (Supelclean Sulfoxide, 3 g/6 mL

SUPELCO), and washed with 4 mL of hexane and the eluate was discarded;

then BDEs were recovered with 10 mL of acetone/hexane (10/90, v/v)

Solvent of the eluate was replaced to hexane and concentrated to 0.2 mL

Analytical instrumentation used GC Agilent GC6890

(e.g., GC-MS ) Injection Splitless 280 deg. 1 uLGC settings Carrier gas He 1.0 mL/min constant flow

(e.g., injection mode, T and volume; carrier gas Column type Frontier Lab. UltraALLOY-PBDE

15 m * 0.25 mmI.D. * 0.05 um Thickness

and flow; column type, oven temperature program) Oven temperature 60 deg. (2 min) - (10 deg./min) - 160deg. (0 min)

(5 deg./min) - 250 deg. (0 min) - (10 deg./min)

- 300 deg. (7 min)MS settings MS Micromass Autospec NT R=10000

(e.g., MS mode, monitored ions, T, electron Mode EI 50 eV SIM

multiplier voltage, gas) Monitor ion 4Br Native: 325.8763, 323.8783, 13C: 337.9171, 335.9191;

5Br Native: 403.7868, 405.7848, 13C: 415.8276, 417.7341;

6Br: 483.6953, 481.6973, 13C: 495.7361, 493.7381

Electron multiplier voltage 350 kV

Water content determination Dried in the oven (105 deg.) for 5 and 7 days.




Calibration type / details IDMS (3-point calibration)



Calibration standards NIST SRM 2257: PBDE Congeners in 2,2,4-Trimethylpentane


Internal standards used EO-5277, CIL ( U.S. EPA Method 1614 Standard Mixtures: including eight

(Please specify the compounds, and at which stage were added) 13C-labeled BDE congeners)

Added to the sample before extraction

14.8 - 16.3



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Extraction method/conditions

Clean-up procedure

(e.g., SPE, GPC)

Analytical instrumentation used(e.g., GC-MS )GC settings

(e.g., injection mode, T and volume; carrier gas

and flow; column type, oven temperature program)

MS settings



Water content determination Procedure as described in protocol




Calibration type / details







BFR-LCS labelled compound solution from Wellington Laboratories was added before extraction. This solution

contain twenty C13 labelled compounds including MBDE-47 (2,2',4,4'-Tetrabromo[13C12]diphenyl ether),

MBDE-99 (2,2',4,4',5-Pentabromo[13C12]diphenyl ether) and MBDE-153 (2,2',4,4',5,5'-

Hexabromo[13C12]diphenyl ether)

Preliminary isotope dilution mass spectrometry followed by bracketing double isotope mass spectrometry

calibration

Calibration solution were used during quantification.

The primary standard, NIST Standard reference material 2257 was used to assign the values to the secondary

standards:

Cambridge Isotope Laboratory, BDE-47-CS-0



The NIST 2257 concentrations are expressed as the certified value ± the expanded uncertainty.

PBDE 47 (2,2',4,4'-Tetrabromodiphenyl ether) - 2.09 ± 0.16 μg/g

PBDE 99 (2,2',4,4',5-Pentabromodiphenyl ether) - 2.127 ± 0.090 μg/g

PBDE 153 (2,2',4,4',5,5'-Hexabromodiphenyl ether) - 2.048 ± 0.068 μg/g

1


GC settings:

The GC-TOFMS system was used for the analysis operating in 1D mode. The injector was set at 300 °C using helium as

the carrier gas at a constant flow of 2.0 ml/min. Samples were injected using a splitless 1µl injection. The GC column

combination consisted of a 5 m x 0.25 mm id Rtx®guard column and a 15 m x 0.25 mm id x 0.1 µm df Rti®5Sil MS

column, connected using a deactivated universal press-tight connector. The GC oven program is provided in table 1.

TOFMS settings:

The transfer line was set at 300°C and the ion source at 250°C. The detector was set at 1800 V to collect a full scan

mass range from 100 – 1000 m/z at a data acquisition rate of 10 spectra/second.

Quantitation and qualification settings:

The quantitation and ions, qualifying ions as well as the and ratios used during the quantification process is listed in

Table 2.

Pressurised Liquid extraction (PLE) settings:

Selective PLE (SPLE) was carried out using a fully automated Accelerated Solvent Extraction (ASE) system. In short,

extraction cells were loaded by inserting two pre-cleaned cellulose filters followed by 5 g of activated neutral

Alumina. The sediment samples were mixed with activated neutral alumina and activated copper powder (1:2:2)

and loaded on the Alumina followed by a celluloses filter. The extraction cell was heated to 100°C and extracted

using two cycles with 3:1 DCM/Hexane as solvent. The total extraction duration was approximately 30 minutes.

The Selective PLE (SPLE) method incorporates in-cell Solid Phase Extraction (SPE) clean-up using activated copper

powder and activated neutral Alumina


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TÜBITAK UME



Sample pre-treatment (if applicable) 2 g of sample was mixed with 5 g of Cu/Na2SO4 (3:1 w/w) mixture and 6 g of

inert dispersing agent and loaded to extraction cell. In order to establish IDMS technique

isotopic labelled standards for all compound were added into the sample at this stage.

Extraction method/conditions Pressurized Solvent Extraction was used. T: 120 C, P:130 bar, Cycle:3

Solvent: n -hexane: acetone (70:30 %), static time 10 min

Clean-up procedure Multilayer column was prepared. From bottom to top 3 g of 3% deactivated

(e.g., SPE, GPC) alumina, 1.5 g of 3% deactivated silica gel, 1.5 g of 44% acidified silica gel and

1 g of Na2SO4 was placed into the glass column. First, the column was washed with

10 mL of n-hexane. Then, 5 mL of extract was loaded. Elution was performed by 50

mL of dichloromethane:n -hexane (1:1) mixture. Final amount was gravimetrically adjusted

to 200 mg by evaporating solvent under gentle stream of nitrogen.

Analytical instrumentation used Triple Quadrupole GC-MS/MS was used. Temperature program was initiated at

(e.g., GC-MS ) 130 ˚C and held for two minutes, increased to 230 ˚C with 5 ˚C/min rate andGC settings increased to 300 ˚C with 5 ˚C/min rate and held for 5 minutes.

(e.g., injection mode, T and volume; carrier gas 15mx0.25mmx0.1µm DB-5MS capillary column was used. PTV injector operating

and flow; column type, oven temperature program) in splitless mode was used. Injection temperature was 150 ˚C (held 0.8 min),

transfer rate, temperature and holding time was 13 ˚C/s, 300 ˚C and 5 min, respectively.

Helium was used as carrier gas at constant flow and 1 mL/min flow rate.MS settings


multiplier voltage, gas) 4712C 485.56 325.84 16

4713C 497.69 337.80 17

9912C 563.72 403.84 15

9913C 575.71 415.94 16

15312C 643.65 483.66 15

15313C 655.64 495.64 15

Water content determination Water determination was carried out by coulometric Karl Fischer titration at 105 ˚C

(please describe the procedure, if deviating in triplicate. Sample intake was 200 mg. Karl Fischer method is treacable to NIST 2890

from the one prescribed in the protocol Water Saturated Octanol standard reference material.


Calibration type / details IDMS single-point calibration was used.



Calibration standards BDE-47, BDE-99, BDE-153 neat crystals were obtained from CHIRON. 13C BDE-47, 13C BDE-99,

(e.g., source, purity, uncertainty) 13C BDE-153 were obtained Wellington Laboratories.

The purity of BD47, BDE-99 and BDE-153 were determined by qNMR and NIST SRM-1944 was

used for further check.

The sample NMR spectrum was given below for BDE-47 in section 7.

Internal standards used 13C BDE-47 13C BDE-99 13C BDE-153

(Please specify the compounds, and at which stage Isotopic labelled compounds were added into the sample at the begining of extraction.

Purity assessment of the calibrant (if applicable) The purity of BDE-47, BDE-99, BDE-153 neat crystals were determined by Q-NMR


Estimation of impurities (if applicable) BDE-47 (99,2±0,03) %

(e.g. type of impurity, mass fraction, uncertainty) BDE-99 (98,7±0,02) %

BDE-153 (99,5±0,05) %

1.12

1.00

1.08


Congener Product Ion (m/z ) Parent Ion (m/z ) CID voltaj Isotope Ratio

2


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VNIIM



Sample pre-treatment (if applicable) No

Extraction method/conditions ASE: Hexane/Acetone=90/10, 20 min, 8 cycles; T=120C; P=1700psi

SoxTherm: Toluene, 16 h, 12 cycles/h

Clean-up procedure As in the Method EPA 1614:

(e.g., SPE, GPC) Silica gel multilayer column

Aluminum oxide, neutral

Copper

Analytical instrumentation used Agilent 6890/5973N

(e.g., GC-MS )GC settings As in the Method EPA 1614:

(e.g., injection mode, T and volume; carrier gas T inj = 300 C, T interface = 320 C

and flow; column type, oven temperature program) Splitless, He, 1 ml/min, Constant Flow

100 C (3 min) - 5 C/min - 320 C (20 min)

MS settings SIM&windows: PBDE#47 - 483,7/485,7 495,7/497,7

(e.g., MS mode, monitored ions, T, electron PBDE#99 - 563,6/565,6 575,7/577,7 PBDE#153 - 641,5/643,5 653,5/655,5

multiplier voltage, gas) DB-5HT 30m*0.25mm*0.10um

Water content determination Mass of samples were about 2 g

(please describe the procedure, if deviating T = 105 C




(e.g., single-point, bracketing / Short range calibration


Calibration standards PBDE Congeners in i-Octane NIST SRM 2257


Internal standards used Method EPA 1614 Labeled Surrogate Stock Solution

(Please specify the compounds, and at which stage EO-5277

2.5



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ANNEX B

Results for BDE 209, optional analyte in CCQM-K102, for which no KCRV

was to be determined.

Table 1-B. BDE209 results

Participant Mass fraction

(µg/kg)

Combined standard

uncertainty u (µg/kg)

Coverage

factor

Expanded

uncertainty U

(µg/kg)

LNE 8751 481 2 962

NIST 7980 220 4.3 944

NIM (GC-MS, NCI) 8776 380 2 760

NIM (HPLC-PAD) 8440 270 2 540

Figure 1-B. BDE209 results, displaying participants' standard uncertainty

7000

7500

8000

8500

9000

9500

NIM (HPLC-PAD) LNE NIM (GC-MS,NCI)

NIST

µg/kg BDE209


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Revised results submitted by some participants in CCQM-K102.

Table 2-B. BDE 47: officially submitted and revised (in yellow) results in CCQM-K102

Participant

Mass

fraction

(µg/kg)

Combined

standard

uncertainty u

(µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Reason for revision

VNIIM 14.8 0.58 (0.65) 2 1.2 (1.3) Slight correction of

uncertainty estimation

LNE 15.77 2.99 (0.87) 2 5.98 (1.74)

mistake in uweighing +

introducing

measurements'

independency

(dividing SD by √n)

TÜBITAK

UME 16.91 (17.72) 0.76 2 1.52 (1.59)

use of 60 m column

(instead of 15 m)

Figure 2-B. BDE 47: KCRV and its standard uncertainty, officially submitted (blue diamonds) and

revised results (yellow squares), displaying participants' standard uncertainty. In red the value

excluded from the KCRV calculation.


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(µg/kg)

Combined

standard

uncertainty

u (µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Reason for revision

NMISA 48.12 (32.21) 4.01 (2.69) 2.025 8.13 (5.44) calculation error

NIST 41.1

(35.0, DB- XLB

34.6, DB-5MS)

0.85 4.35

3.7

(2.90, DB- XLB

2.14, DB-5MS)

using DB-XLB and

DB-5MS 30 m

columns (instead of

10 m) to improve

separation of potential

interferences

LNE 30.93 3.83 (1.095) 2 7.66 (2.19)

mistake in uweighing +

introducing

measurements'

independency

(dividing SD by √n)

TÜBITAK

UME 39.50 (35.24) 2.04 2 4.07 (3.63)

Use of 60 m column

(instead of a 15 m) to

improve separation of

potential interferences



Figure 3-B. BDE 99 KCRV and its standard uncertainty, officially submitted (blue diamonds) and

revised results (yellow squares), displaying participants' standard uncertainty. In red the value(s)


N.B. The revised NIST results displayed are the ones obtained using the 30 m DB-5MS column.


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(µg/kg)

Combined

standard

uncertainty

u (µg/kg)

Coverage

factor

Expanded

uncertainty

U(µg/kg)

Reason for revision

NMISA 7.12 (7.29) 0.52 (0.54) 2.013 1.05 (1.09) calculation error

TÜBITAK

UME 11.03 (6.86) 0.72 2 1.45 (0.90)

Use of 60 m column

(instead of 15 m) to

improve separation of

potential interferences

NIST 8.97

(6.10, DB-XLB

6.50, DB-5MS)

0.2 4.35

0.87

(0.55, DB-XLB

0.65, DB-5MS)

using DB-XLB and

DB-5MS 30 m

columns (instead of

10 m) to improve

separation of potential

interferences

LNE 7.03 1.75 (0.46) 2 3.50 (0.92)

mistake in uweighing and

introducing

measurements'

independency by

dividing SD by √n



Figure 4-B. BDE 153 KCRV and its standard uncertainty, officially submitted (blue diamonds) and

revised results (yellow squares), displaying participants' standard uncertainty. In red the value(s)


N.B. The revised NIST results displayed are the ones obtained using the 30 m DB-5MS column.


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ANNEX C

Full details of the uncertainty budgets estimated by the participants in CCQM-K102

BAM

Contributions to the uncertainty budget and worst case estimation

ucom=SQRT(Ʃui2). The individual contributions ui are as follows:

-------------------------------------------------------------------------------------------------------------------------------

CENAM

m0 Standard mass

mI0 Standard isotope mass

mx Sample mass

mIx Standard isotope mass in sample

R0 Response ratio of standards

Rx Response ratio of sample

w0 Mass fraction of standard

CFh Humidity correction factor

wx Mass fraction of measurand in sample

10

)(

ilod

losspuritychex

samplealiquot

issolventislod

sample

sample

Fandcorwith

FFFFmm

cmmcor

sl

icrc

CFhwmRm

mRmw

Ix

xIx

x *0

00

0


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GLHK

Each standard uncertainty of PBDE- 47, 99 and 153 was obtained by combining the systematic

uncertainty and random uncertainty as shown below equation.

The systematic uncertainty included the followings:

1) the uncertainty of calibration standard solution (Ucal) by combining the uncertainty of preparation of

standard solution (weighing) and uncertainty of NIST BDE standard solutions obtained from the

certificate;

2) the uncertainty due to preparation of sample blend and calibration blend (Uweighing);

3) the uncertainty due to moisture correction (Umoisture)

The random uncertainty was calculated from the precision (standard deviation) of multiple measurement

results from three bottles.

-------------------------------------------------------------------------------------------------------------------------------

JRC-IRMM

k coverage factor (k=2) resulting in a confidence level of approx. 95 %

u cal relative uncertainty related to the preparation of the calibration standards

including contribution arising from purity and gravimetric steps

RSD rep relative standard deviation of repeatability (from ANOVA) - from validation data

n1 number of independent samples used for analysis (n1=7)

RSD ip relative standard deviation of intermediate precision (from ANOVA) – from validation data

n2 number of days (n2=1)

u true relative uncertainty of trueness, estimated from the recovery - from validation data

------------------------------------------------------------------------------------------------------------------------

22

2

2

1

2

* caltrue

iprepuu

n

RSD

n

RSDkU

u rep, % uip, % u trueness, % ustds, % u, % U, %

BDE47 3.3 1.3 0.9 3.8 4.3 8.6

BDE99 7.4 8.5 3.3 2.1 9.8 19.5

BDE153 6.7 11.9 2.8 1.7 12.6 25.2

MoistureR

R

M

M

M

MCC

Bc

B

Yc

Zc

X

YZX


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KRISS

f: dry-mass correction factor;

Csample: is the concentration of analytes in the sample;

Cs-sol: is the concentration of the analytes standard solution;

Msample: is the mass of the sample taken for analysis;

Mis-sol, spiked: is the mass of the isotope standard solution added to the sample aliquot;

Mis-sol, std. mix.: is the mass of the isotope standard solution added to the isotope ratio standard solution;

Ms-sol, std. mix.: is the mass of the standard solution added to the isotope ratio standard solution;

ARsample: is the area ratio of analyte/isotope for sample extract, observed by GC/MS;

ARstd. mix.: is the area ratio of analyte/isotope for the isotope ratio standard solution, observed by GC/MS.

Random : Standard deviations of multiple measurement results from five subsamplings

Combined standard uncertainties were obtained by combining systematic uncertainties and random uncertainties as shown below equation

------------------------------------------------------------------------------------------------------------------------

n

suCu

22

systematics.p.,mean )(


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LGC

LNE

Csed: mass fraction of PBDE in the sediment in µg/kg Clab: mass fraction of labeled PBDE solution in µg/kg mlab: mass of labeled solution in the sediment

msed: mass of sediment sample Rsed: unlabeled/labeled ion peak area ratio in the sediment sample

a: gradient of the slope for linear regression plot b: intercept on y axis for the linear regression plot

f standard: correction factor due to the standard solutions uncertainty

f F: correction factor due to measurement precision

Fdards

sed

ffbRsedam

mlabClabCsed

tan)(


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PBDE 47

Type (A or B) relative Uncertainty (%)

vial 8 vial 44 vial 77

preparationof sample blends (weighings) B 60.10% 14.64% 73.65%

Calibration model B 0.11% 0.03% 0.15%


B 15.15% 3.75% 21.26%

Precision B 24.65% 81.57% 4.95%

PBDE 99






B 47.69% 42.18% 51.76%

Precision B 8.69% 18.25% 1.67%

PBDE 153






B 8.17% 0.50% 7.98%

Precision B 5.48% 92.38% 12.37%

------------------------------------------------------------------------------------------------------------------------

NIM

Measurement uncertainty mainly come from measurement precision and calibration solution.

Uncertainty source BDE-47 BDE-99 BDE-153 BDE-209

（HPLC)

BDE-209

（GCMS)

Mesurement

result(µg/kg) 14.65 35.00 7.177 8.440 8.776

Method precision 0.18 0.33 0.11 0.26 0.37

Calibration solution 0.29 0.70 0.14 0.059 0.061

Combined standard

uncertainty 0.35 0.78 0.19 0.27 0.38

Coverage factor 2 2 2 2 2

Combined expanded

uncertainty 0.70 1.56 0.38 0.54 0.76

------------------------------------------------------------------------------------------------------------------------


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NIST

The combined uncertainty was estimated as the mean value times the square root of the sum of

squares of the relative uncertainties (CVs) for: triplicate measurements, water content

determination, calibration and the uncertainty in the certified value of the calibration solution. The

95 % expanded uncertainties were estimated as the two-tailed Student’s t value for two degrees of

freedom, t(95,2) = 4.3, times the combined uncertainty

------------------------------------------------------------------------------------------------------------------------

NMIA

where;

ωx = mass fraction of analyte in sample

ωz = mass fraction of analyte in the calibration standard solution used to prepare calibration blend

My = mass of internal standard solution added to sample blend

Myc = mass of internal standard solution added to calibration blend

Mx = mass of sample added to sample blend

Mzc = mass of calibration standard solution added to calibration blend

Rb = ratio of measured peak areas corresponding to the analyte and internal standard in the sample blend

Rbc = ratio of measured peak areas corresponding to the analyte and internal standard in the calibration blend

Rx = ratio of measured peak areas corresponding to the analyte and internal standard in the sample

Rz = ratio of measured peak areas corresponding to the analyte and internal standard in the calibration standard

Ry = ratio of measured peak areas corresponding to the analyte and internal standard in the internal standard

MCCF = moisture content correction factor

F(MP) = method precision term

F(MT) = method trueness factor

All masses and mass fractions used to calculate ωx were determined using balances calibrated

with metrological traceability to the SI unit of the kilogram through Australian national standards for mass.

Peak areas were determined from chromatographic traces generated for characteristic ions corresponding to analytes and

internal standards. 32 sub-samples taken from Bottles 21, 37, 53 and 81 of the study material were analysed in

seven batches between November 2014 - March 2015. Moisture content was determined according to the study protocol.

Tables showing the measurement uncertainty budgets for BDEs 47, 99 and 153 are provided on the Tabs named,

Uncertainty BDE47, "Uncertainty BDE99", and "Uncertainty BDE153".

A standard uncertainty was estimated for all components in the measurement equation. These were combined using

derived sensitivity coefficients to estimate a combined standard uncertainty in the reported result for each analyte

in the study sample. The total effective degrees of freedom was determined using the Welch-Satterthwaite

equation to calculate the appropriate coverage (k) factor to expand the combined standard uncertainty to a 95% confidence

interval for reporting. The method precision term was calculated as the standard deviation of all the measurements used in the

calculation of the reference value.

To ensure that all likely sources of bias would be accounted for in the final uncertainty budget a trueness factor was also included.

This factor was assigned a nominal value of one with an uncertainty representing the potential magnitude of undetected bias due to

factors affecting the measured peak area ratios such as matrix interferences and matrix effects and bias due to the incomplete

recovery of native BDE analytes from the sediment matrix. The magnitude of the standard uncertainty in the trueness factor was

estimated as the standard deviation of the various average analyte mass fractions determined in multiple sediment samples

analysed by the primary method of analysis and the different confirmatory methods of analysis used to assess bias due to matrix

interferences/effects and the recovery of analytes from the sample matrix.

MTMP

BcY

ZcB

XB

BY

Yc

Zc

X

YZX FFMCCF

RR

RR

RR

RR

M

M

M

M


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BDE 47

BDE 99

Summary of Contributions to Total Combined Measurement Uncertainty

Number Name of Component Symbol Units Value

Standard

Uncertainty

Relative

Standard

Uncertainty

Degrees of

Freedom

i Xi xi u(xi) u(xi)/xi (%) vi

1 Method Precision F(MP) dimensionless 1.000 0.031 3.1% 40.0

2 Method Trueness F(MT) dimensionless 1.000 0.018 1.8% 7.0

3 Standard Wz ng/g 328 14 4.4% 6.0

4 Moisture Content MC n/a 1.00502 0.00093 0.092% 5.0

5 Gravimetry Mx g 2.0126 0.00026 0.013% 100.0

6 Gravimetry My(SB) g 0.0869 0.00026 0.30% 100.0

7 Gravimetry Mz g 0.0944 0.00026 0.28% 100.0

8 Gravimetry My(CB) g 0.0852 0.00026 0.31% 100.0

9 Isotope Amount Ratio Rx,Rz mol/mol 6.1E+05 5.3E+05 87% 4.0

10 Isotope Amount Ratio Ry mol/mol 0.000012 0.000021 186% 4.0

11Blend Isotope Amount

RatioR(SB) mol/mol 0.7878 25.0


RatioR(CB) mol/mol 0.7614 25.0


Number

Name of

Component Symbol Units Value

Standard

Uncertainty

Relative Standard

Uncertainty

Degrees of

Freedom


1Method

PrecisionF(MP) dimensionless 1.000 0.049 4.9% 40.0

2Method

TruenessF(MT) dimensionless 1.000 0.017 1.7% 7.0

3 Standard Wz ng/g 792 31 3.9% 6.0

4Moisture

ContentMC n/a 1.00502 0.00093 0.092% 5.0

5 Gravimetry Mx g 2.0126 0.00026 0.013% 100.0


7 Gravimetry Mz g 0.0883 0.00026 0.29% 100.0


9Isotope Amount

RatioRx,Rz mol/mol 3.2E+06 1.6E+06 49% 4.0

10Isotope Amount

RatioRy mol/mol 0.0000087 0.0000032 37% 4.0

11Blend Isotope

Amount RatioR(SB) mol/mol 1.1672 25.0

12Blend Isotope

Amount RatioR(CB) mol/mol 1.1886 25.0


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BDE 153

---------------------------------------------------------------------------------------------------------------------------------

NMIJ


Number Name of Component Symbol Units Value

Standard

Uncertainty

Relative

Standard

Uncertainty

Degrees of

Freedom


1 Method Precision F(MP) dimensionless 1.000 0.044 4.4% 36.0

2 Method Trueness F(MT) dimensionless 1.000 0.021 2.1% 6.0

3 Standard Wz ng/g 126.5 3.4 2.7% 6.0

4 Moisture Content MC n/a 1.00502 0.00093 0.092% 5.0

5 Gravimetry Mx g 2.0130 0.00026 0.013% 100.0


7 Gravimetry Mz g 0.0911 0.00026 0.29% 100.0


9 Isotope Amount Ratio Rx,Rz mol/mol 5.8E+06 1.4E+06 24% 1.0

10 Isotope Amount Ratio Ry mol/mol 0.000000163 0.000000037 22% 4.0


RatioR(SB) mol/mol 1.0950 22.0


RatioR(CB) mol/mol 0.9936 22.0


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BDE47 total No.11 No.39 No.73

concentration of primary standard

0.038 0.038 0.038 0.038

mass ratio of standards solutions

0.00022 0.00022 0.00022 0.00022

mass ratio of sediment and 13C-BDE

0.00011 0.00011 0.00011 0.00011

Water content 0.000088 0.000088 0.000088 0.000088


0.0048 0.0046 0.0033 0.004

calibration curve 0.0095 0.0094 0.01 0.0092

total 0.04 0.04 0.04 0.04

N.B. total uncertainty of "repeatability from GC/MS" was calculated from ANOVA

BDE99 total No.11 No.39 No.73

concentration of primary standard

0.021 0.021 0.021 0.021

mass ratio of standards solutions

0.00022 0.00022 0.00022 0.00022

mass ratio of sediment and 13C-BDE

0.00011 0.00011 0.00011 0.00011

Water content 0.000088 0.000088 0.000088 0.000088


0.0021 0.0038 0.0085 0.0095

calibration curve 0.013 0.013 0.013 0.013

total 0.024 0.025 0.026 0.026


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------------------------------------------------------------------------------------------------------------------------

NMISA

BDE 47


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BDE 99

BDE 153

------------------------------------------------------------------------------------------------------------------------

TÜBITAK UME

Bottom up approach was used considering the following sources:

x u u/x u/x2 v

W z

[native] solution added to calibration blend

(ug/ g)68208.45 1936.70 0.03 8.06E-04 2

m z

weight native solution added to calibration

blend (g)0.155 1.00E-04 6.44E-04 4.15E-07 5

m y

weight of Isotope solution added to sample

(g)0.099 1.00E-04 1.01E-03 1.01E-06 5

m yc

weight of Isotope solution added to

calibration blend (g)0.099 1.00E-04 1.01E-03 1.02E-06 3

m x weight of sample analysed 1.000 1.00E-04 1.00E-04 9.99E-09 5

R' B /R' BC

ratio of peaks areas of native/ labelled in the

samples1.088 4.19E-02 3.85E-02 1.48E-03 5

Precision Repeat measurements 32.334 2.06E+00 6.37E-02 4.06E-03 5

FRepeat

dry mass correction factor1.004 2.36E-02 2.35E-02 5.55E-04 7

2.69 u

5.44 U (k = 2.013)

16.83 Urel

Factors in uncertainty

x u u/x u/x2 v

W z

[native] solution added to calibration blend

(ug/ g)66454.14 1875.79 0.03 7.97E-04 2

m z

weight native solution added to calibration

blend (g)0.155 1.00E-04 6.44E-04 4.15E-07 5

m y

weight of Isotope solution added to sample

(g)0.099 1.00E-04 1.01E-03 1.01E-06 5

m yc

weight of Isotope solution added to

calibration blend (g)0.099 1.00E-04 1.01E-03 1.02E-06 3

m x weight of sample analysed 1.000 1.00E-04 1.00E-04 9.99E-09 5

R' B /R' BC

ratio of peaks areas of native/ labelled in the

samples1.032 4.22E-02 4.09E-02 1.67E-03 5

Precision Repeat measurements 7.080 3.49E-01 4.93E-02 2.43E-03 5

FRepeat

dry mass correction factor1.004 2.36E-02 2.35E-02 5.55E-04 7

0.52 u

1.05 U (k = 2.013)

14.87 Urel

Factors in uncertainty


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1-Mass of sample intake

Value Standard Uncertainty

Mass of sediment sample

Calibration msediment umcalibrationsample

Mass of Tare

Calibration mtare umcalibrationtare

Since same balance was used umcalibrationsample and umcalibrationtare is equal to same value

2-Isotopic Labelled Compounds Stock Solution


Purity of Compound

PCompound13C12 uPCompound13C12

Mass

Mass of first compound

Calibration m1st compound umCalibration

Mass of second compound

Calibration m2nd compound umCalibration

Mass of third compound

Calibration m3rd compound umCalibration

Mass of Solvent

Calibration msolvent umCalibration

Mass of Tare

Calibration mtare umCalibration

Since same balance was used all umcalibration are equal to same value

Combined Standard Measurement Uncertainty

3-Spiking of Isotopic Labelled Stock Solution by Gas Tight Syringe


Calibration

uVC

Temperature

uVT


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4-Uncertainty of Recovery

uCobs standard measurement uncertainty of observed concentration of analyte

Cobs observed concentration of analyte

uCcert standard measurement uncertainty of certified concentration of analyte

Ccert certified concentration of analyte

5-Uncertainty of Repeatability

6-Karl-Fisher Water Determination - Mass of Sample

- Repeatability of Karl Fisher measurement

7-Mass of final sample


Mass of sediment final sample

Calibration mfinal sample umcalibrationsample

Mass of solvent

Calibration mtare umcalibrationsolvent

Mass of Tare

Calibration mtare umcalibrationtare

Since same balance was used all umcalibration are equal to same value

------------------------------------------------------------------------------------------------------------------------------


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VNIIM

-----------------------------------------------------------------------------------------------------------------------------

uA – standard uncertainty of results of 9 measurements, % ;

uB – standard uncertainty consisting of

standard uncertainty of calibration – ucal, %,

standard uncertainty of weighing of sample – usample, %,

standard uncertainty of blank – ublank, %;

and standard uncertainty of dry mass determnation – udry, %.

In turn ucal consists of

standard uncertainty of SRM – usrm, %,

standard uncertainty of weighing of solution – usolution, %,

standard uncertainty of spiking of IS – uis, %,

and of standard uncertainty of RR (relative response) – uRR, %.

The meanings of u sample, u blank, u dry are negligible so the equation is


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ANNEX D

KCRVs, associated expanded uncertainty and the participants' results

displaying expanded uncertainties for BDE 47, BDE 99 and BDE 153 in

CCQM-K102.

Figure 1-D. CCQM-K102: KCRV and its expanded uncertainty for BDE 47.

Participants' results are also displayed with their expanded uncertainties.

KCRV (median) 15.60

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 1.19


U95 % (k*u) k = 2 0.82


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KCRV (median) 33.69

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 2.15


U95 % (k*u) k = 2 1.62



KCRV (median) 6.28

𝑀𝐴𝐷𝑒 = 𝑚𝑎𝑑 ∗ 1.483 0.74


U95 % (k*u) k = 2 0.56


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ANNEX E

CCQM-K102: Core Competency Tables [Analyte(s) in Matrix]

Instructions:

In the middle column place a tick, cross or say the entry is not applicable for each of the

competencies listed (the first row does not require a response)

Fill in the right hand column with the information requested in blue in each row

Enter the details of the calibrant in the top row, then for materials which would not meet the CIPM

traceability requirements the three rows with a # require entries

BAM

CCQM-K102 BAM PBDE in sediment Scope of Measurement: quantification of organic molecules in the approximate range of molecular weights

from 100 to 800 g/mol, having polarity corresponding to pKow < -2 and for the range of mass fraction 1-1000

μg/kg in abiotic dried matrices.

Competency

Tick,

cross,

or

“N/A”

Specific Information as Provided by

NMI/DI

Competencies for Value-Assignment of Calibrant

For calibrants which are a highly-pure

substance: Value-Assignment / Purity

Assessment method(s).#

N/A

For calibrants which are a calibration

solution: Value-assignment method(s).#

N/A

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time, mass spec ion ratios, (GC/MS)

Extraction of analyte(s) of interest from

matrix √ ASE, toluene, 3 cycles, 100°C, 140 bar

Cleanup - separation of analyte(s) of

interest from other interfering matrix

components (if used)

√ Clean-up

1) Al2O3 column chromatography

(1 cm of sodium sulfate; 25 g of Al2O3; 1 cm of

sodium sulfate)

2) Multilayer column chromatography

(1,0 g of sodium sulfate / 0.5 g silica gel/silver

nitrate; 0.2 g of silica gel;

1.0 g of silica gel/sulfuric acid; 0.2 g of silica gel;

0.5 g of silica gel/sodium hydroxide)

Transformation - conversion of analyte(s)

of interest to detectable/measurable form (if

used)

N/A none

Analytical system √ GC-MS, quadrupole

Calibration approach for value-assignment

of analyte(s) in matrix √ a) IDMS, internal standards labelled BDE 47,99,153

b) 6-point calibration curve

Verification method(s) for value-

assignment of analyte(s) in sample (if used)

N/A

Other N/A

BDE 153 degree of equivalence expanded uncertainty does not cross zero, value not consistent

with the KCRV: no specific competency identified as reason.


of 82

CENAM

CCQM-K102 NMI PBDEs in sediment Scope of Measurement: quantification of organic molecules in the approximate range of molecular weights


μg/kg in abiotic dried matrices.

Competency

Tick,

cross,

or

“N/A”


NMI/DI


Calibrant: Did you use a “highly-pure

substance” or calibration solution?

Calibration solution from Cambridge Isotope

Laboratories (CIL) verified by SRM2257

Identity verification of analyte(s) in

calibration material.#

√ GC-MS/MS






√ We use SRM-2257from NIST to verify calibrants RF,

CIL calibration curve were verified with SRM2257

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time and MRM ion pairs


matrix √ Automated Soxhlet Extraction




√ SPE silica

Transformation - conversion of analyte(s) of

interest to detectable/measurable form (if

used)

N/A

Analytical system √ GC-MS/MS


of analyte(s) in matrix √ IDMS single-point calibration

Verification method(s) for value-assignment

of analyte(s) in sample (if used)

N/A SRM 2257 was used as control RM which were treated

under the same sample prep as the test sample

Other N/A Indicate any other competencies demonstrated.


of 82

GLHK

CCQM-K102 GLHK Track „A“- low polarity analytes in abiotic matrix

Polybrominated diphenyl ethers in sediment

Scope of Measurement: Quantification of organic molecules in the approximate range of MW from 100 to 800

g/mol, having polarity corresponding to pKow < -2 and for the range of mass fraction 1 – 1000 µg/kg in abiotic

dried matrices.

Competency

Tick,

cross, or

“N/A”


NMI/DI




Certified reference material SRM 2257 (PBDE

congeners in 2,2,4-trimethylpentane) obtained from the

NIST.



N/A




N/A



N/A

Sample Analysis Competencies Identification of analyte(s) in sample √ Relative retention time, ion ratios


matrix √ Soxhlet extraction

Cleanup - separation of analyte(s) of interest

from other interfering matrix components (if

used)

√ Multi-silica gel column and alumina column clean-up



used)

N/A

Analytical system √ GC-HRMS (Double focusing HRMS)

Calibration approach for value-assignment of

analyte(s) in matrix √ a) IDMS

b) exact matching



N/A

Other


of 82

JRC-IRMM

CCQM-K102

JRC-

IRMM

Polybrominated diphenyl ethers in

sediment Scope of Measurement: quantification of organic molecules in the approximate range of MW from 100 to 800

g/mol, having low polarity (pKow < -2) and for the range of mass fraction 1 – 1000 µg/kg in abiotic dried

matrices

Competency

Tick,

cross, or

“N/A”


NMI/DI




Calibration solution NIST SRM 2257



Cross-checked by GC-MS with in-house prepared

standards from neat crystals




N/A



N/A

Sample Analysis Competencies Identification of analyte(s) in sample

Retention time, mass spec ion ratios by GC-MS


matrix

ASE




SPE



used)

N/A

Analytical system

GC-ECNI-MS


of analyte(s) in matrix

a) internal standard (PBDE 77)




Value assignment cross checked with in-house

prepared standards and quality control samples

(spiked study sediment and SRM 1944, with reference

mass fraction values)

Other Method was in-house validated previously


of 82

KRISS

CCQM-K102 KRISS


sediment Scope of Measurement: quantification of organic molecules in the approximate range of molecular weights


μg/kg in abiotic dried matrices

Competency

Tick,

cross, or

“N/A”


NMI/DI




Neat commercial calibrants for BDE-47, 99, 153 were

from Accustandard.



√ GC/MS




√ The purities were assessed by KRISS with GC-FID, but

water content and non-volatile impurities were not

tested due to the limited amounts of the materials.

Purities: BDE 47, 99, and 153 were 98.75%, 95.50%,

and 95.83%, respectively. The purities assessed by

KRISS was indirectly confirmed by comparing the

calibration solution prepared with the materials to NIST

SRM 2257



√ Calibration solutions were gravimetrically prepared in

KRISS and verified by cross-checking of multiple

calibration solutions. Secondary confirmation by

comparison with NIST SRM 2257

Sample Analysis Competencies Identification of analyte(s) in sample √ GC Retention time, mass spec ion ratios, comparison of

GC/MS measurement results by low resolution SIM,

high resolution SIM at two different channels for each

analytes


matrix √ Pressurized Liquid Extraction (PLE) with

dichloromethane




√ SPE cartridge (silica gel)



used)

N/A

Analytical system √ On-column GC/MS (low resolution) in SIM mode at a

primary channel and a secondary (confirmatory)

channel for each analytes


of analyte(s) in matrix √ IDMS with exact matching single-point calibration


assignment of analyte(s) in sample (if used) √ On-column GC-HRMS (R=10000) for confirmatory

measurement

Other N/A


of 82

LGC

CCQM-K102 LGC


sediment Scope of Measurement: Demonstration of the lab's competency in quantification of organic

molecules in the approximate range of MW from 100 to 800 g/mol, having polarity

corresponding to pKow < -2 and for the range of mass fraction 1 – 1000 µg/kg in abiotic dried

matrices.

Competency

Tick,

cross,

or

“N/A”


NMI/DI




Pure solid materials of PDBEs 47, 99 and 153 were

obtained from Chiron. Their identity was confirmed

within LGC by 1H NMR and MS.



The calibration solutions of the three calibrants in

2,2,4-Trimethylpentane were prepared in house from

pure materials above




N/A PDBEs 47 and 99 ‘pure materials’ were value assigned

by qNMR using maleic acid as an internal standard (

maleic acid purity assigned in house by LGC)



√ 2,2,4-trimethylpentane stock solutions of the three

PDBEs were prepared from ‘pure’ solid materials and

then assayed by qNMR using dimethyl sulphone

(DMSO2) as an internal standard in deuterated

acetonitrile. Purity of DMSO2 reference material

assigned by LGC and NMIJ.

Sample Analysis Competencies Identification of analyte(s) in sample √ Comparison with standards in terms of Retention Time

and full scan spectra, as well as ion ratios between SIM

quantifier and qualifier ions


matrix √ Exhaustive soxhlet extraction, with acetone/hexane

solvent composition




√ Initial clean-up of the extracts was with SPE(acid/basic

florisil columns) followed by normal phase LC

fractionation, using hypercarb columns with a

toluene/hexane mobile phase and temperature gradient



used)

N/A N/A

Analytical system √ GC-MS with EI source, equipped with Dean switch used

as divert valve


of analyte(s) in matrix √ Bracketing double exact matching IDMS was used for

all three analytes with 13C internal standards



N/A

Other N/A


of 82

LNE

CCQM-K102 LNE Polybrominated diphenyl ethers in

sediment Scope of Measurement: demonstrate the laboratories competence in the quantification of organic

molecules in the approximate range of molecular weights from 100 to 800 g/mol , having polarity

corresponding to pKow < -2 and for the range of mass fraction 1-1000 µg/kg in abiotic dried matrices.

Competency

Tick,

cross,

or

“N/A”


NMI/DI




SRM2257


calibration material.# √

Identification by comparison with the pure analyte , (

mass spectrum ,retention time, abundance of

characteristics ions) and with the NIST library




N/A



N/A

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time and specific transitions


matrix √ ASE



used)

√ Purification of the ASE extract on a pretreated silica

and alumina column



used)

N/A Indicate chemical transformation method(s), if any, (i.e.,

hydrolysis, derivatization, other)

Analytical system √ GC/MS²


of analyte(s) in matrix √ a) quantification mode used : IDMS

b) calibration mode used: 11 points calibration curve



N/A Indicate any confirmative method(s) used, if any.

Other N/A Indicate any other competencies demonstrated.


of 82

NIM

CCQM-K102 NIM


sediment Scope of Measurement: quantification of organic molecules in the approximate range of MW from 100

to 800 g/mol, having polarity corresponding to pKow < -2 and for the range of mass fraction 1 – 1000

µg/kg in abiotic dried matrices.

Competency

Tick,

cross,

or

“N/A”


NMI/DI



substance” or calibration solution? GBW(E)081124 , GBW08709, from NIM,China.



√ GC-MS




√

The purity of neat BDE-47,99,153 is determined by

HPLC-DAD and qNMR. The purity of BDE- 209 is

determined by HPLC-DAD and HPLC-ICP-MS



√ The calibration solution is prepared by the weight

capacity method.

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time and mass spec ion ratios


matrix √

ASE



used) √ SPE



used) N/A No

Analytical system √ 1) BDE-47,99,153: GC-MS/MS

2) BDE-209: HPLC-PDA


of analyte(s) in matrix √

a) BDE-47,99,153: IDMS

b) BDE-209: internal standard.

c) single-point calibration,


of analyte(s) in sample (if used) √

Recovery is determined by adding standard solution to

blank sediment.

Other “N/A”




of 82

NIST

CCQM-K102/P138 NIST Polybrominated Diphenyl Ethers in Sediment Scope of Measurement: quantification of organic molecules in the approximate range of MW from 100 to 800

g/mol, having polarity corresponding to pKow < -2 and for the range of mass fraction 1-1000 µg /kg in abiotic

dried matrices

Competency Specific Information as Provided by NMI/DI




NIST Standard Reference Materials 2257 and 2258


calibration material. √ Preparation from commercially obtained materials,

confirmed by GC-MS

For highly-pure substance: Value-

Assignment / Purity Assessment method(s). NA

For calibration solution: value-assignment

methods. √ Gravimetric preparation and analytical results

determined using gas chromatography.

Sample Analysis Competencies Identification of analyte(s) in sample √ Identification in samples based on retention time and

MRM transitions obtained with SRM 2257 and 2258


matrix √

Pressurized Fluid Extraction



used)

√ Elution through 1.8 g SPE cartridge of deactivated

alumina.



used)

NA

Analytical system X GC-MS/MS

For BDE 99 and 153 results biased high, interferences

not separated during chromatography


of analyte(s) in matrix √

Isotope dilution with calibration curve.


of analyte(s) in sample (if used) √ Concurrent analysis of PBDE congeners in SRM 1944

New York New Jersey Waterway Sediment.

BDE 99 and 153 degree of equivalence expanded uncertainties do not cross zero (see Analytical

system box above), values not consistent with the KCRV.


of 82

NMIA

CCQM-K102 NMIA

Low Polarity Analytes in an Abiotic

Matrix: Polybrominated Diphenyl

Ethers in Sediment Scope of Measurement:

Quantification of organic molecules in the approximate range of molecular weights from 100 to 800 g/mol,

having polarity corresponding to pKow < -2 and for the range of mass fraction 1-1000 µg /kg in abiotic

dried matrices.

Competency

Tick,

cross, or

“N/A”


NMI/DI




Calibration solutions of BDEs 47, 99 and 153

Source: Wellington Laboratories, Ontario, Canada

Product numbers: BDE-47, BDE-99 and BDE-153



- Chromatographic retention time

- GCHRMS – a minimum of 2 ions monitored

- GCMSMS – a minimum of 2 SRM

transitions monitored

- LCMSMS – a minimum of 2 SRM





N/A N/A



Multiple standards comparison against solution

matrix certified reference materials, NIST SRM 2257

and NIM China GBW (E) 081124 and GBW (E)

081125

Sample Analysis Competencies Identification of analyte(s) in sample

- Chromatographic retention time

- GCHRMS – a minimum of 2 ions monitored

- GCMSMS – a minimum of 2 SRM


- LCMSMS – a minimum of 2 SRM



matrix Method 1: Accelerated Solvent Extraction (ASE)

with toluene

Method 2: Hot Soxhlet extraction with toluene

Method 3: ASE with ethanol/toluene (68:32) and

toluene

Method 4: ASE with acetone/hexane (50:50)



used)

1. Concentrated sulphuric acid wash

2. Ultra-high purity water wash

3. Solid phase extraction with multi-layer (acidic,

neutral and basic) silica and alumina columns

Note: Gel permeation chromatography (GPC) with

dichloromethane was used as an additional clean-up

step for SRM1944, a matrix reference material for

PBDEs in sediment. GPC clean-up was found to be

unnecessary for the CCQM study sample.


of 82



used)

N/A N/A

Analytical system GC-HRMS (EI)

GC-MS/MS (EI) (for confirmation)

LC-MS/MS (APCI/APPI) (for confirmation)

Calibration approach for value-assignment of

analyte(s) in matrix Exact-matching (single-point calibration) double

isotope dilution mass spectrometry with

bracketing


of analyte(s) in sample (if used) Comparison of results using independent extraction

and detection techniques

1. ASE with ethanol/toluene and toluene

2. ASE with acetone/hexane

3. Hot Soxhlet extraction with toluene

4. GC-MS/MS (EI) analysis

4. LC-MS/MS (APCI/APPI) analysis with high

performance liquid chromatography (HPLC) clean-up

of sediment extracts

5. GCHRMS analysis using chromatography columns

with stationary phases of differing polarity (J&W

DB5 and Agilent VF-17MS)

Other 1. Investigation of complete analyte/internal standard

equilibration

- comparison of experimental procedures involving

addition of internal standard directly to sediment

samples packed in ASE cells versus addition of

internal standard to sediment samples weighed into

glass vials and then mixed by rotation for a minimum

of 12 hours

- exhaustive extraction of sediment samples

(optimisation of the number of static cycles and the

static/hold time used for ASE)

- comparison of Hot Soxhlet extraction of sediment

samples for 10 hours versus ASE

- comparison of different extraction solvents for ASE

2. Investigation of possible background

contamination: analysis of method blank samples,

using pre-cleaned hydromatrix as the blank matrix


of 82

NMIJ

CCQM-K102 NMIJ PBDEs in sediment Scope of Measurement: Organic molecules in the approximate range of molecular weights from 100 to 800

g/mol, having low polarity corresponding to pKow < -2 and for the range of mass fraction 1-1000 µg /kg in

abiotic dried matrices.

Competency

Tick,

cross, or

“N/A”


NMI/DI




Calibration solution (NIST SRM 2257)



GC/MS (The BDE-47, 99, and 153 was identified

from the information on retention order of BDE

isomers from used GC column (IEC 62321, 2008),

and comparison to another standard solution, EO-

5277 (CIL).)




N/A -



NIST certificate (Purity assessment by GC and

Gravimetric preparation)

Sample Analysis Competencies Identification of analyte(s) in sample GC/MS (Retention time and mass spec ion ratios)


matrix

Ultrasonic extraction (CH2Cl2)




Sulfur cleanup with activated Cu

Sulfuric acid treatment (column packed with silica

impregnated with concentrated sulfuric acid, 44 %)

SPE (Supelclean Sulfoxide, 3 g/6 mL; SUPELCO)


of interest to detectable/measurable form

(if used)

N/A -

Analytical system GC-HRMS (EI, R=10000) equipped with GC

column, UltraALLOY-PBDE (15 m x 0.25 mmI.D. x

0.05 um Thickness)


of analyte(s) in matrix

a) IDMS



assignment of analyte(s) in sample (if

used)

x (The extraction and cleanup conditions have been

optimized using NIST SRM 1944. However, obtained

analytical results for tetra- to hexa-BDEs have not

been verified.)

Other N/A




of 82

NMISA

CCQM-K102 NMISA Polybrominated diphenyl ethers in

sediment Scope of Measurement: Demonstration of the competency in quantification of organic molecules in the

approximate range of Mw from 100 to 800 g/mol, having polarity corresponding to pKow < -2 and for the

range of mass fraction 1 – 1000 µg/kg in abiotic dried matrices.

Competency

Tick,

cross,

or

“N/A”


NMI/DI



substance” or calibration solution? Calibration solution.



√ Verification against NIST SRM 2257 through mass

spectral ion ratios and retention times using Gas

Chromatography – Time-of-flight mass spectrometry

(GC-TOFMS)




N/A



√ The primary standard, NIST Standard reference

material 2257 was used to assign the values to the

secondary standards:

Cambridge Isotope Laboratory, BDE-47-CS-0, BDE-

99-CS-0 and BDE-153-CS-0. Value assignment was

performed using external and isotope dilution mass

spectrometry (IDMS) calibration

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time

Mass spectrometric ion ratios

Full range mass spectra


matrix √ Pressurized liquid Extraction (PLE)




√ Solid phase extraction (SPE)

Sulphuric acid digestion and activated copper cleanup



(if used)

N/A

Analytical system √ Gas Chromatography – Time-of-flight mass

spectrometry (GC-TOFMS)


of analyte(s) in matrix √ Preliminary IDMS followed by bracketing double

IDMS calibration



used)

N/A

Other √ Water content determination by gravimetric loss on

drying as specified in protocol


with the KCRV: calculation error.


of 82

TÜBITAK UME

CCQM-K102 NMI:TÜBİTAK UME PBDEs in sediment Scope of Measurement: Demonstration of the lab's competency in quantification of organic molecules in the

approximate range of MW from 100 to 800 g/mol, having polarity corresponding to pKow < -2 and for the

range of mass fraction 1 – 1000 µg/kg in abiotic dried matrices.

Competency Tick, cross,

or “N/A”


NMI/DI




BDE-47, BDE-99, BDE-153 neat crystals were

obtained from CHIRON. (1962.125MG, 1967.12-

5MG, 1971.12-5MG) 13

C BDE-47, 13

C BDE-99, 13

C BDE-153 were

obtained from Wellington Laboratories.

(MBDE471009, MBDE0990611, MBDE1530710)

The purity of BD47, BDE-99 and BDE-153 were

determined by qNMR and NIST SRM-1944 was used

for further check, Newyork/New Jersey Waterway

Sediment



Triple quadrupole GC-MS/MS




qNMR


solution: Value-assignment

method(s).#

N/A -

Sample Analysis Competencies

Identification of analyte(s) in sample

IDMS, GC-MS/MS

Extraction of analyte(s) of interest

from matrix

Pressurized Solvent Extraction was used. T: 120 ˚C,

P:130 bar, Cycle:3, Solvent: n-hexane: acetone

(70:30 %), static time 10 min




Multilayer column was prepared. From bottom to top

3 g of 3% deactivated alumina, 1.5 g of 3%

deactivated silica gel, 1.5 g of 44% acidified silica

gel and 1 g of Na2SO4 was placed into the glass

column. First, the column was washed with 10 mL of

n-hexane. Then, 5 mL of extract was loaded. Elution

was performed by 50 mL of dichloromethane:

n-hexane (1:1) mixture. Final amount was

gravimetrically adjusted to 200 mg by evaporating

solvent under gentle stream of nitrogen

Transformation - conversion of

analyte(s) of interest to

detectable/measurable form (if used)

N/A -

Analytical system X Triple Quadrupole GC-MS/MS

For BDE 99 and 153 results biased high,

interferences not separated during chromatography

Calibration approach for value-

assignment of analyte(s) in matrix

IDMS single-point calibration



used)

Recovery were examined by using NIST SRM-1944,

Newyork/New Jersey Waterway Sediment and spiking

of pure compounds to matrices for double check.

Other N/A -

BDE 99 and 153 degree of equivalence expanded uncertainties do not cross zero (see

Analytical system box above), values not consistent with the KCRV.


of 82

VNIIM

CCQM-K102 VNIIM


sediment Scope of Measurement: Quantification of organic molecules in the approximate range of MW from 100 to 800 g/mol, having low

polarity (pKow < -2) and for the range of mass fraction 1 - 1000 µg/kg in abiotic dried matrices

Competency

Tick,

cross, or

“N/A”

Specific Information as Provided

by NMI/DI




PBDE Congeners in i-Octane NIST SRM 2257



N/A




N/A



N/A

Sample Analysis Competencies Identification of analyte(s) in sample √ Retention time

Ion ratios of the two molecular m/z


matrix √ SoxTherm (Toluene, 16 h, 12 cycles/h)

ASE (Hexane/Acetone=90/10, 20 min, 8 cycles;

T=120 ºC; P=1700 psi)




√ Silica gel clean-up (multilayer column with Sil,

Sil/H2SO4, Sil/KOH)

Al2O3 fractionation

Copper clean-up



(if used)

N/A

Analytical system √ GC-LRMS


of analyte(s) in matrix √ a) IDMS (mass of IS – about 40 ng)

b) Calibration into short range (in accordance with

analytes mass in the sample – from 16ng to

74ng)



used)

N/A As the reference meaning was taken average result

of long-time (48h) Soxhlet extractions

Other N/A

CCQM-K102 Polybrominated diphenyl ethers in sediment...CCQM-K102_Final report.doc E-mail:...

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