analysis of environmental contaminants using high performance quantitative lc/ms/ms - waters...

©2015 Waters Corporation 1

Analysis of Environmental Contaminants using High

Performance Quantitative LC-MS/MS


Overview of the presentation

Background environmental contaminants – EU legislation overview

– POPS (dioxins; PCBs; PFCs; pesticides), endocrine disruptors

Environmental LC-MS/MS analysis – the challenges?

Why use mass spectrometry? – Waters MS tandem quadrupole solutions

Multi-residue pesticide analysis in drinking water

Direct quantification of acidic herbicides in water

Strategies for the analysis of Eds & POPs

Perfluorinated compounds (PFCs)

Summary


Transformation products

Drinking water (targeted analysis)

Perfluorinated compounds

(PFCs)

Environmental applications

Marine studies

Environmental water

Unknown Analysis

Dioxins POPs

Endocrine disrupting

compounds


Legislation – contamination & environmental factors

Regulations on the use of specific chemical substances, e.g. those

used in farming or in certain production or food processing techniques

– Measures are taken to limit pollution of water or air, or from exposure to

radioactivity

EU – Drinking Water Directive (98/83/EC)

– EU Marine Strategy Framework Directive (2008)

– EU Water framework Directive (2010)

– Community strategy for endocrine disruptors SEC (2011) 1001

EU “criteria based” approach adopted

– Water Framework Directive methods must be certified according to EN

ISO/IEC-17025

USA

– Drinking Water Regulation

– WHO Guidelines Drinking water quality (2011, 4th edition)

USA “EPA approved methods of analysis”

http://water.epa.gov/scitech/drinkingwater/labcert/analyticalmethods.cfm#approved


Inhalation

Ingestion

POPs

How do pollutants get into the environment?


Why are Persistent Organic Pollutants (POPs) a problem?

Persistence These compounds do not easily break down into simpler, less harmful forms. They remain and continue to build up in our environment.

Toxicity Dioxins have been proven to be highly toxic in both animal and human

studies e.g. chloracne, cancer, learning difficulties, cognitive impairment,

Health Impacts High exposures to dioxins have been directly related to cancer. Low level, prolonged exposure

has been linked to a host of effects including hormonal and immune system dysfunctions, birth

defects, miscarriages & learning disabilities.

Bioaccumulative Dioxins are stored in fat tissue and continue to accumulate throughout

our lifetime. The body has difficulty metabolizing dioxins, especially

laterally substituted congeners.


Environmental LC-MS/MS – challenges?

Can I detect all of the

target compounds and

what about unknowns?

Do my results comply with

the regulatory

requirements?

Detection capability

Confirmation

Productivity Ease of use

Accessibility

Can my staff use

the technology?

Cal 10 inj1 before 10x dilution

Time0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00

%

0

100

Sample throughput

Can I analyse enough

compounds/samples?


Why use Mass Spectrometry (MS)?

Mass spectrometers offer a number of advantages;

– Measure specific analytes in the presence of thousands of others,

simultaneously (multi-residue analysis)

– Measure trace quantities of analytes (femtomolar concentrations)

– Provide structural information about an analyte

– And can do all of these things at the same time

“The 3 S’s”

– Selectivity; differentiating between the components in a sample

– Specificity; measuring only analytes in the presence of many other

compounds

– Sensitivity; detection of femtomolar concentrations


Tandem quadrupole - instrument anatomy


Tandem quadrupole MS Versatility and power

Tandem quadrupole instruments offer many modes of operation

Each of the quadrupoles can operate in Scanning or Static Modes

– Selected Ion Recording (SIR)

– Precursor Ion Scanning (PAR)

– Constant Neutral Loss (CNL)

– Product Ion Scanning (DAU)

– Multiple Reaction Monitoring (MRM) or Selective Reaction Monitoring

(SRM)

4-5 orders linear dynamic response

Switch between positive and negative ion modes in the same

chromatographic experiment

Meet the legislative requirements for analyte confirmation

Easy to use & robust measurements


Single Ion Recording (SIR)

MS1

Static (m/z 821.5)

Precursor(s)

Q1 Static m/z 821

Collision

Cell (OFF)

Static (m/z 821.5)

Quadrupole 2

Precursor(s) Precursor(s)


SIR


Multiple Reaction Monitoring (MRM),

m/z 821 >768

Analyte Injected in Matrix Hypothetical Example

0.50 1.00 1.50Time0

100

%

0

100

%

0.50 1.00 1.50Time0

100

%

0

100

%

3ng / L 30ng / L

Selected Ion Recording (SIR),

m/z 821


Product Ion Spectrum

790 795 800 805 810 815 820 825 830 835 840 845 850m/z0

100

%

821.5

810.5

822.5

826.5

827.5

200 250 300 350 400 450 500 550 600 650 700 750 800 850 900m/z0

100

%

768

576

558548718

750

786

821

Q1 Static m/z 821

Q2 Scanning


Multiple Reaction Monitoring (MRM)

Quadrupole 1 Collision

Cell

Static (m/z 821.5)

Static (m/z 768.5)

Ar (2.5 – 3.0e-3mBar)

Precursor(s) Product(s)

Quadrupole 1


MRM


Multiple Reaction Monitoring (MRM),

m/z 821 >768

Analyte Injected in Matrix Hypothetical Example

0.50 1.00 1.50Time0

100

%

0

100

%

0.50 1.00 1.50Time0

100

%

0

100

%

3ng / L 30ng / L

Selected Ion Recording (SIR),

m/z 821


Solutions for Targeted Screening Analysis

Xevo TQD • Entry level

• UPLC compatibility

• Multi-residue methods

Xevo TQ-S micro

• Increased sensitivity

• Increased speed

• PICs

• RADAR

• Universal source

Xevo TQ-S • Ultimate sensitivity


The Universal Source – configurations

and options


MS Adoption: UPLC-Xevo TQD

Intermediate: UPLC- Xevo TQ-S micro

Max efficiency: UPLC-Xevo TQ-S MS

Less Sample Prep More

Solutions for Targeted Screening The need for sample preparation?


Ultra-trace analysis Waters MS solutions

Ultra trace analysis How can Xevo TQ-S help?

Xevo TQ-S performance enables:

Reduced volumes during sample prep and analysis

Proactively monitor matrix levels during analysis

Improved reproducibility due to increased signal intensity

Better quality results, minimal user intervention


• Automatically sets up instrument,

• Optimises MRMs and performs system suitability

IntelliStart

• Methods generated automatically by QuanPedia,

• Clear flagging when data is out of regulatory tolerance

• Reduced risk of errors

TargetLynx

• Long term system monitoring, Lab management tool

TrendPlot

• Generates optimised methods, time scheduled MRMs

Quanpedia

• Real time QC, Intelligent ‘on the fly’ decision making

QCMonitor

Waters quantitative workflow and software solutions


Multi-residue pesticide analysis in drinking

water

in accordance with 98/83/EC


Prepare sample

• Dechlorinate

• Transfer to sample vial

Analyse 100 µL aliquot

• UPLC separation

• Xevo TQ-S detection

Review results

• Achieve required limits

• Reproducible ppq detection

Direct injection of drinking water Within the EU Drinking Water Directive (DWD) sets quality standards for drinking water quality at the tap (microbiological, chemical and organoleptic parameters • 0.1μg/L (100 ppt) for each individual pesticide in drinking water • 0.5μg/L (500 ppt) total Pesticides (the sum of all the substances detected in a sample)

• stricter separate health based standards for 4 organochlorine pesticides which are no longer

permitted to be used

Drinking water analysis


Time4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

How does StepWave™ help?

Time4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

4.00 4.25 4.50 4.75 5.00 5.25 5.50 5.75 6.00 6.25 6.50 6.75 7.00

%

0

100

Enhanced sensitivity

with StepWaveTM

ion

optics

Linuron

Azinphos- methyl

Atrazine

Metosulam

Without

StepWaveTM ion

optics

100µL drinking water, 81 compounds, 10 x below EU limits (10 ppt)


Ultra trace analysis Direct injection of drinking water

200 pg/L (200ppq) pesticides in drinking water

1.8 ng/L (1.8 ppt) total pesticides

Acetamiprid 223>126

223>56

Omethoate 214>125

214>183

Thiabendizole 202>175

202>131

Simetryn 214>124

214>96

Hexazinone 253>171

253>71

Terbuthiuron 229>172

229>116

Metalaxyl 280>220

280>192

Pirimiphos - methyl

306>164

306>108

Chlorpyriphos 350>198

350>97

Detection capability


Measurement repeatability

Pesticide classes:

Summary of

repeatability @ 100ppt

Ametryn 1.39 Terbutryn 1.96 Cyanazine 1.26 Atrazine 1.46 Simetryn 1.78 Spiroxamine 1.85 Kresoxim Methyl 4.29 Azoxystrobin 2.19 Dimethomorph 4.14 Pyraclostrobin 4.18 Chlortoluron 0.59 Siduron 1.24 Monuron 1.56 Monolinuron 1.09 Diuron 1.24 Dicrotophos 0.94 Heptenophos 1.47 Mevinphos 2.34 Tetrachlorvinphos 2.47 Chlorfenvinphos 3.67 Omethoate 1.22 Demeton S Methyl 1.50 Azinphos Methyl 2.48 Dimethoate 2.27 Ethoprophos 2.31

2.04 Mean

Class Compound %RSD (n=32)

Triazine herbicides

Organothiophosphorous pesticides

Fungicides

Phenylurea herbicides

Organophosphorous pesticides


TrendPlot

TrendPlot report – reproducibility @100 ppt (EU limit)

%RSD 0.83 Fenuron

%RSD 0.78 Simazine

%RSD 0.71 Flumeturon


River water

•A relatively simple matrix?

Total Organic Carbon content (TOC) can commonly be between 0.5 to 10 ppm

• If 30 compounds are present at 100 ppt (EU limit) this is 3 ppb of detected compounds

If we have 5 ppm TOC, and detect 3 ppb of targeted compounds, what else is in the sample?

•Food is a much more complex sample

MRM is targeted, we see only what we tell the instrument to show us

•How can we tell what is also present?

Monitoring the matrix effects in river water


Rapid electronics allow instrument to

switch between MRM and full scan (FS

takes 100ms)

– No loss of MRM data quality

– Added information can be gained from

full scan

o Monitor matrix

o Assess for possibility of matrix effects

o Added information for sample prep

method development

o Search for significant non-targeted

compounds

RADAR (Xevo TQ-S & Xevo TQ-S micro) Simultaneous full scan & MRM

MRM

Full scan 81 compound multi-residue

method, using RADAR


100 ng/L (100 ppt) pesticides in river water

RADAR view – what else is in the sample?


PCP XIC (m/z 264.8)

TIC

m/z 264.8 - PCP

RADAR - TIC

Sewage Effluent spiked with 20 ng/L pentachlorophenol (PCP)


Strategies for the analysis of endocrine

disruptors &

Persistent organic Pollutants (POPs)


Endrocrine disrupting compounds Natural hormones & synthetic compounds

Time1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

1003.13

3.193.10

3.09

2.97

2.46

17a-ethinylestradiol

17b- and 17a-estradiols

Estrone

Bisphenol A

Estriol

Endocrine

Disruptors

Challenging LODs

8 minute analysis

17b- and 17-a-estradiols

Estrone

Bisphenol A

Estriol


Endrocrine disrupting compounds

Endocrine

Disruptors

10ng/ml matrix matched standard

Time1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

100

1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

%

0

1003.06

3.133.03

3.01

2.81

2.35

17a-ethinylestradiol

17b- and 17a-estradiols

Estrone

Bisphenol A

Estriol

Compounds

Method LOD ng/L 200 mL sample

Estriol 4

Bisphenol A 2

Estrone 0.5

17-estradiol 1

17-ethinyl estradiol 1.5

17-estradiol 1

17b- and 17-a-estradiols

Estrone

Bisphenol A

Estriol


Perfluorinated compounds (PFCs)

perfluoroalkyl sulfonates (PFAS), perfluoroalkyl carboxylates

(PFAC), perfluorohexane sulfonic acid (PFHxS) & perfluorooctane

sulfonic acid (PFOS)


Xevo TQ-S Perfluoronated compounds (PFCs)

Projects to determine the levels of PFCs in

– Mink Liver

– Fish Liver

– Drinking Water

PFCs present in the environment after use of fire fighting foams

The production, supply, use and disposal of PFOS in Europe is controlled by the Persistent Organic Pollutants Regulation (EC) 850/2004 Contaminating groundwater or surface water with firewater that contains PFOS is not permissible At present there is no lower concentration limit in the EU POPs Regulations –any material containing any concentration of PFOS should be disposed of or recovered by one of the prescribed methods

Evaluation of UPLC with Xevo TQ-S

– Instrument sensitivity

– Explore RADAR Acquisition

– Reduction of matrix effects via sample dilution


PFCs analysis using Xevo TQ-S Fire fighting foams

Seepage water samples from fire training activity

after cleaning steps (more complex sample)

Borehole from lake adjacent to fire training activity at an

airport in a different location (less complex sample)


Instrument sensitivity PFCs 200fg on column

PFBA

PFPeA

PFHpA

PFHxA

PFOA

PFNA

PFDA

PFDoDA

PFUnDA

PFTrDA

PFBuS

PFHxS

PFOS

PFDS


Impact of sensitivity RADAR data for complex samples

Original Sample

x10 dilution

x100 dilution

acquisitions allow levels of matrix to be monitored at the same time as running MRM experiments

Other matrix components can affect both chromatography AND ionization (ion suppression)


PFHxA

x100

x10

x 2

.5

decre

ase

x 1

0.2

decre

ase

802268

318971

31270

Peak Area

Reducing matrix (dilution)

Original Sample (Seepage sample close to FFF source)

x10 dilution

x100 dilution

Expected RT


PFHxA (Peak Area vs Matrix Conc)

0

500000

1000000

1500000

2000000

2500000

3000000

3500000

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1

Relative Matrix Concentration

Peak A

rea

Ion s

uppre

ssio

n

Undiluted sample

~75%


Summary

Monitoring environmental contamination and the advantages of mass spectrometry? – Fundamentals and inside the mass spectrometer

– How does all of this work?

– Multiple Reaction Monitoring, RADAR and new ion source designs

The application of tandem quadrupole instruments to environmental analytical applications – Multi- residue pesticide analysis in drinking water

– Acidic herbicides

– Surface water analysis

– Endocrine disruptors

– POPs and perfluorinated compounds

Sensitivity is not just about detecting lower concentrations of analytes; it’s also about being able to dilute out interferences

Mass Spectrometry has become more accessible and routine than ever before

analysis of environmental contaminants using high performance quantitative lc/ms/ms - waters...

Science

waters corporation

eu water framework directive

water quality

water strategies

pollution of water

usa drinking water regulation

bioaccumulative dioxins

toxicity dioxins