A d v an cin g P BB an d P BD E A dv an cin g P BB an d P ... · PDF file5 ©2007 Waters...

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©2007 Waters Corporation

Advancing PBB and PBDE Advancing PBB and PBDE Analysis In Compliance To Analysis In Compliance To RoHSRoHS

Using UPLCUsing UPLCTMTM/MS/MS

©2007 Waters Corporation 2

OutlineOutline

• Overview of RoHS and WEEE.

• Overview of PBBs and PBDEs.

• Sample Preparation for PBB/PBDE Analysis.

• Analysis of PBB/PBDE using UPLCTM/ZQ.

• Savings and Advantages

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Overview of Overview of RoHSRoHS and WEEEand WEEE

• The Restriction of the Use of Certain Hazardous Substances (RoHS) Directive : 2002/95/EC

• The RoHS regulations ban the putting of new Electrical and Electronic Equipment (EEE) containing more than the permitted levels of lead (Pb), cadmium (Cd), mercury (Hg), hexavalent chromium (Cr6+) and both polybrominatedbiphenyl (PBB) and polybrominated diphenyl ether (PBDE)flame retardants on the EU market from 1 July 2006.


• Waste Electrical and Electronic Equipment (WEEE) Directive : 2002/96/EC

• WEEE aims to minimise the impact of EEE on the environment throughout their life cycle. Criterias are set on collection, treatment, recycling and recovery of waste. These activities are financed by producers and such waste can be returned by consumers without charge. This comes in force with effect from 13 August 2005.

Overview of Overview of RoHSRoHS and WEEEand WEEE

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Purpose of RoHS

• To reduce harmful / hazardous substances at source.

• Maximum concentration values established through the adoption of Commission Decision 2005/618/EC where the following will be permitted in the manufacturing of new EEE.

Pb, Hg, Cr6+, PBB, PBDE : Up to 0.1% weight

Cd : Up to 0.01% by weight.

Purpose of WEEE

• WEEE : To minimise the impact of these wastes on the environment by promoting the reuse, recycling and other forms of recovery of such waste.

Overview of Overview of RoHSRoHS and WEEEand WEEEPurposePurpose

homogeneous material


Overview of Overview of RoHSRoHS and WEEEand WEEEImplementation Implementation

Implementation

• Producers are responsible for ensuring that the products do not contain the mentioned hazardous substances.

• Compliance can be demonstrated by submitting technical documentation or other information.

• Perform analysis to ensure products comply with the maximum concentration values.

• Producers must ensure that they understand and take into account any limitations of the analytical technique used.

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Overview of Overview of PBBsPBBs

• PBB = Polybrominated Biphenyls

• Used as flame retardants, added to plastics used in products such as home electrical appliances, textiles, plastic foams etc. to make them difficult to burn.

Chemical Structure of PBBChemical Formula : C12H(10-n-m)Brn+m(both n and m = 1 to 5)

10 substitutions (209 congeners)

(1) Monobromobiphenyl

(2) Dibromobiphenyl

(3) Tribromobiphenyl

(4) Tetrabromobiphenyl

(5) Pentabromobiphenyl

(6) Hexabromobiphenyl

(7) Heptabromobiphenyl

(8) Octabromobiphenyl

(9) Nonabromobiphenyl

(10) Decabromobiphenyl

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Overview of Overview of PBBsPBBs

• Most of the PBB congeners found in the commercial flame retardants are

(i) lipophilic – it can be stored under fats.

(ii) persistent - reach the environment sooner or later because of the high stability of these compounds.

(iii) bioaccumulate - posing a particular threat to organisms in the higher levels of these webs

• Forms toxic polybrominated dibenzofurans during combustion processes.

• Shown to produce chronic toxic effects and cancer.


Overview of PBDEOverview of PBDE

• PBDE = Polybrominated Diphenyl Ethers

Chemical Structure of PBDEChemical Formula : C12H(10-x-y)Brx+yO

(both x and y = 1 to 5)

10 substitutions (209 congeners)

(1) monobromodiphenyl ethers

(2) dibromodiphenyl ethers

(3) tribromodiphenyl ethers

(4) tetrabromodiphenyl ethers

(5) pentabromodiphenyl ethers

(6) hexabromodiphenyl ethers

(7) heptabromodiphenyl ethers

(8) octabromodiphenyl ethers

(9) nonabromodiphenyl ethers

(10) decabromodiphenyl ethers

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Overview of PBDEOverview of PBDE

• Used as additive flame retardants which are incorporated into the plastic matrix like other additives, such as plasticizers.

• High-impact polystyrene, ABS, flexible polyurethane foam, textile coatings (not clothing), wire and cable insulation, electrical/electronic connectors and other interior parts.

• Polybrominated dibenzofurans (PBDF) and polybrominateddibenzodioxins (PBDD) can be formed from PBDEs, under different conditions, including heating (combustion).

• Hundreds of possible congeners of halogenated dibenzofurans and dibenzo-dioxins. Of these hundreds congeners, substituents in the 2,3,7,8-positions are toxic.







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Sample PreparationSample Preparation

Reference :

Procedures for the determination of levels of regulated

substances in electrotechnical products’ developed by the

International Electrotechnical Committee (IEC) ACEA ad

hoc working group.

• Sample : Homogeneous material

• Cutting :

(1) Not more than 2x10x10 cm3.

(2) Electronics : 4x4cm

(3) Polymer : 5x5mm Coarse Grinding

Homogenizing

Fine Grinding


Sample PreparationSample Preparation

• 100mg of samples (grain size of 1mm)

• Undergoes soxhlet extraction for 3 hours using 70ml of n-propanol.

• Filter the samples.

• Make up to 100ml.

• Additional concentration steps or higher sample mass required if the concentration is close to the detection limit.

Other alternatives such as ultrasonic extraction has known to be adopted.

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• Savings and Advantages.


What is UPLC?What is UPLC?UUltra ltra PPerformance erformance LLiquid iquid CChromatographyhromatography

A new class of separation science— Based on chromatography columns with very small particles

— Based on instruments holistically designed to take advantage of the small particles

UPLCUPLC™™ isis Particle Size Driven

Requires a system which can sustain very high pressure to attain the linear velocities required to achieve the resolution enabled by small particles, and maintain that resolution through to detection

Provides improved Resolution, Speed, and Sensitivity

Suitable for chromatographic applications in general— Appropriate for improving existing methods

— Appropriate for developing new methods

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Ultra Performance LCUltra Performance LCTMTM

Resolution,Resolution, Speed and SensitivitySpeed and Sensitivity

0.25

AU

Minutes0.0 1.6

Speed: 9X (900%) faster separations

Minutes

AU

0.000

0.010

0.020

0.030

0.040

0.050

3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00

Sensitivity: up to 3X (300%) increase

Resolution: 1.7 (70%) increase

AU

0.00

0.02

0.04

0.06

0.08

0.10

Minutes0.00 5 10 15 20 25 30 35 40 45 50 55 60


New 2New 2ndnd Generation HybridGeneration HybridACQUITY UPLCACQUITY UPLCTMTM ChemistryChemistry

Tetraethoxysilane(TEOS)

Bis(triethoxysilyl)ethane(BTEE)

+4

Polyethoxysilane(BPEOS)

Si

EtO

EtO OEtEtO

Si

EtOEtO

CH2EtO

CH2Si

OEt

OEtOEtSi

EtO

O

CH2 CH2

Si O

Si

EtO

OEt

Si O

O

OEtO

Si

O

Si

OEt

O

OOEt

Et

Et

n

• Improved Strength• Improved Efficiencies• Improved Peak Shape • Wider pH Range

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ACQUITY UPLCACQUITY UPLCTMTM

Customer Column Lifetime StudyCustomer Column Lifetime Study

• Isocratic aging experiment— ACQUITY UPLC™ BEH C18 (2.1 x 50 mm, 1.7 mm)— Low pressure run (initial data used to benchmark column

efficiency)o 0.5mL/min, ~6,000 psi

— High pressure runs of 400 injectionso 1.25mL/min. ~13,300 psi

— This aging cycle was repeated and tested every 400 injections

— Low pressure runs/injections efficiency at 1600/4000 (data shown)

• Columns lasted over 4,000 high pressure injections— At this point study was abandoned with satisfactory results

"After 4000 injections I took the column off. I was convinced."

Customer quote following the evaluation.


First Injection on Column No high pressure runsN = 3483 for Peak 1 N = 6352 for Peak 2

Injection after1600 high pressure runs

N = 3659 for Peak 1N = 6489 for Peak 2

Injection after4000 high pressure runs

N = 3573 for Peak 1N = 6466 for Peak 2

AU

- 0 . 4 0

- 0 . 2 0

0 .0 0

0 .2 0

0 .4 0

M in u t e s0 .0 0 0 .2 0 0 . 4 0 0 .6 0 0 .8 0 1 . 0 0 1 .2 0 1 .4 0 1 . 6 0 1 .8 0 2 .0 0 2 . 2 0 2 .4 0 2 .6 0 2 . 8 0 3 .0 0

1.16

5

1.97

4

AU

-0.40

-0.20

0.00

0.20

0.40

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00

1.16

8

1.97

8

AU

-0.40

-0.20

0.00

0.20

0.40

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00

1.17

8

2.00

0

Ketorolac Naproxen

Chromatograms obtained under low (~6000 psi) isocratic conditions

ACQUITY UPLCACQUITY UPLCTMTM

Results ofResults of Customer Column Lifetime StudyCustomer Column Lifetime Study

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ACQUITY UPLCACQUITY UPLCTMTM TechnologyTechnologyUltra performance by designUltra performance by design

Detectors:Optical and/or Mass SpecTunable UV or Photodiode ArrayOptimized flow cell for UPLCTM

High speed data sampling

Column Manager:Innovative pivot designUp to 65oCPositions column to detectorE-CordTM connectionSample Manager:

Low dispersion XYZZ’ FormatVariable vol. fixed loop injectorLow volume injectionsFast cycle timesDual wash - Low carryoverPlates and/or vialsThermal control (4-40oC)Pressure tolerant

Binary Solvent Manager:High pressure blendingBinary gradientsFour solvent choicesOn-line degassingLow dispersion designUPLC pressure capabilities

Sample Organizer: (option)Expands capacity (22/15/8)Shuttles plate feedHeated/chilled

System Considerations:Small FootprintRedesigned tubing and fittingsConsolidated waste managementIntegrated system diagnosticsConnections InsightTM remote diagnostics

Holistic Design


New Single Quad Detection for UPLCNew Single Quad Detection for UPLC

SQD Detector designed to be:

— UPLC Compatible

— Smaller in size

— Multi-mode

— Easier to operate

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Waters SQ DetectorWaters SQ DetectorSmallest tandem quadrupole on the benchSmallest tandem quadrupole on the bench

34cm (~13”) of linear bench space


Waters SQD Mass Spectrometer Waters SQD Mass Spectrometer

Flexibility

Ionization mode: dedicated ESI or APCI

Standard ESCi probe: combined ESI/APCI

Optional APPI source

Fast scan speeds up to 10.000 amu/sec

Fast polarity Pos/Neg switching 20 ms

Fast Inter Channel Delay 5ms for SIR

Mass range: 0 -2000 amu

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Accuracy and Stability

Extreme long term mass stability by new electronics

Robustness and Sensitivity

Z Spray source

Tool free routine maintenance

Data Acquisition Up to 32 functions per sample Full scan or SIR MassLynx and Empower full software

control

Waters SQD Mass Spectrometer Waters SQD Mass Spectrometer


UPLCUPLC™™ and MS Detectionand MS DetectionAdvantages over traditional HPLCAdvantages over traditional HPLC

UPLCä and MS— Improved ion intensity

— Reduced ion suppression

Low system volume— Fast gradients and re-equilibration

— Low peak dispersion – high resolution

— Quick method changes

Minimal injection carry-over— High sensitivity applications

Fast injection cycle time— High throughput

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Developed in parallel with UPLC™ to be used with UPLC— SQDTM

— TQDTM

— Quattro Premier™— LCT Premier™— Q-Tof Premier™

Used for many assays— Quattro micro™— LCT ™— Q-Tof™— Q-Tof micro™

UPLCUPLC™™ and MS Detectionand MS DetectionAvailable MS DetectorsAvailable MS Detectors


Analysis of PBDEAnalysis of PBDEHPLCHPLC--UVUV--MS ChromatogramMS Chromatogram

Mass Spectrometer Chromatogram

UV Chromatogram

Isomers of technical Octa-PBDE and Deca-PBDE are assigned. Signals between 3.5 to 8 minutes could not be identified as one of the flame retardants investigated in this study.- Abstracted from J. of Chromatography A, 1064 (2005)

Sample of shredded monitor housing

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Analytical Parameters For UPLCAnalytical Parameters For UPLCTMTM

• A1 : 0.1% formic acid / H2O

• B1 : 0.1% formic acid / ACN

• Isocratic separation with A1/B1 = 15/85

• Flow rate : 0.6ml/min

• BEH C18 (2.1mm x 100mm x 1.7mm).

• Column Temp. : 50°C

• 1ul injection volume with PLUNO mode

• PDA detector : 200~500nm at Rs=1.2nm and sampling rate 40Hz.


Preliminary Analysis of PBDEPreliminary Analysis of PBDEUPLCUPLCTMTM ChromatogramChromatogram

Deca-BDE

Nona-BDE

Nona-BDE

Octa-BDE(DE197)

Octa-BDE(DE196)

Hexa-BDE(DE153)

Hexa-BDE(DE154)

1ul injection

1000ppm DE79 (Octa-BDEs)

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UPLC ChromatogramUPLC Chromatogram

Nona-BDE

Deca-BDE1ul injection

1000ppm DE83u (Deca-BDEs)

Preliminary Analysis of PBDEPreliminary Analysis of PBDEUPLCUPLCTMTM ChromatogramChromatogram


Preliminary Analysis of PBDE Preliminary Analysis of PBDE UPLCUPLCTMTM/ZQ/ZQ

10ppm of Octa-PBDE, Nona-PBDE and Deca-PBDE

Octa-PBDE

Deca-PBDENona-PBDE

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Preliminary Analysis of PBDE Preliminary Analysis of PBDE UPLCUPLCTMTM/ZQ/ZQ

1ppm of Octa-PBDE, Nona-PBDE and Deca-PBDE

Octa-PBDE

Nona-PBDE Deca-PBDE






• Savings and Advantages.

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Savings and AdvantagesSavings and Advantages

• Run time per sample using HPLC or GC = 20mins

• Run time per sample using UPLC = 5mins

• Higher throughput.

• Reduced amount of eluent used.

• Faster turnaround time.

• UPLCTM/ZQTM promises confirmatory results.

4x faster

A d v an cin g P BB an d P BD E A dv an cin g P BB an d P ... · PDF file5 ©2007 Waters...

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Transcript of A d v an cin g P BB an d P BD E A dv an cin g P BB an d P ... · PDF file5 ©2007 Waters...