The world leader in serving science

Sandro Spezia, Enzo Trivellone

Modena 29 Maggio 2009

Q-ICP-MS Mass

Spectrometry: new

challenges in food safety and

environmental applications

2

XSERIES 2

XSERIES 2 CCT ED

Q ICP-MS

Q ICP-MS CCTED

ELEMENT 2

ELEMENT XR

High Resolution ICP-MS

Neptune

Multicollector ICP-MS

Se

lectivity

Thermo Fisher Bremen, Germany

Noise: 0.2 cpsNoise: 0.2 cps

Center of Excellence for ICP-MS

3

Introduction to the XSERIES 2 ICP-MS

Bench top quadrupole ICP-MS

Efficient Ar plasma ion source

Capable of detecting sub-pg mL-1

levels across most of the Periodic Table

Large dynamic range

• Can measure from sub-pg mL-1 to

mg mL-1 in a single run

Offers collision cell technology for

removing polyatomic interferences

Versatile technology, but ..........

• ....... limited to samples containing

< 0.2% (m/v) dissolved solids

(cones block above this level)

4

Interface cone options - what to use and when?

Xt interface

• Ni skimmer cone mounted in Cu-Ni adapter

• Gives low backgrounds for cone derived

analytes (Ni, elements thermally ionised from

the cones)

• Provides lower polyatomic interference levels

e.g. ArO+ and ArH+ on Fe and K than standard

(Xs) cones

• Provides ideal mass response for ppm

Na, K, Mg, Ca measurement with ppt

detection for trace heavy metals

• Offers good long term stability for

analysis, but shows significant signal

suppression for higher matrix samples

Na: blank to 300 ppm Cd: blank to 100 ppb

5

Application Specific Interfaces – Xs-

Extraction lens operated at

negative potential

• High sensitivity (>200kcps/ppb)

• Low background noise (<0.5 cps)

Highest signal to noise of any

quadrupole ICP-MS

Applications:

• Environmental (radionuclides),

isotope ratio, laser, GC/LC…

6

Example of Xs-: High Sensitivity U Determinations

ppb

7

Nebuliser options - what to use and when?

Glass concentric nebuliser

• standard design for the XSeries

• sample uptake rate ~ 0.8 mL min-1 (free aspirated

or pumped)

• good all-round performance (sensitive, stable),

but can block with higher salt samples

Burgener Miramist

• parallel path nebuliser design (liquid channel

alongside gas channel)

• sample uptake rate ~ 0.8 mL min-1 (pumped only)

• resistant to blocking; good for biological samples

Low flow concentric nebulisers (PFA)

• sample uptake rates down to 20 mL min-1 (free

aspirated mainly)

• susceptible to blocking

• best nebuliser for semicon analysis or low sample

volume work with filtered samples

8

XSERIES 2 Schematic

9

What is a CRC ?

A multipole enclosed in a cylinder

Controlled flow of gas into the cell

Interaction of ions with the gas

If reactive gas used, reactions occur

• All cells are reaction cells

Collision/Reaction Cell Technology

M+ and

XnYn‟+

M+ only out

10

Basic Schematic of CRC ICP-MS

Gas 1 Gas 2

MFC 1 MFC 2

Detector

Quadrupole

Pre-

LensMultipole CellPost-

Lens

Rotary Pump

Plasma

Split-flow High-compression Turbo

11

(1) Collision-induced dissociation

Ar-Ar++ He Ar + Ar+ + He

Not thought to contribute much

12

(2) Chemical reaction

ArAr+ + H2 ArH+ + ArH

A major mechanism for removal

13

(3) Electron transfer

ArAr+ + H2 ArAr + H-H+

A major mechanism for removal

14

(4) Collisional retardation/differential transmission

* ArAr+ + He ArAr+ + * He

*

*

a.k.a kinetic energy discrimination

15

CCTED - Kinetic Energy Discrimination for Collisional or Reactive Chemistry

Collisional retardation / energy filtering

Collision/Reaction gas atom or

molecule

Analyte Ion M+ - Small collision

cross-section

Polyatomic Species e.g. ArX+ –

Larger collisional cross-section

Post-Cell Cell Pre-Cell

Decre

asin

g E

nerg

y

Energy

Barrier

From plasma

To quadrupole π Extraction Lens –

ensures that the energy of

the ions entering the cell are

ideal for collisional or

reactive chemistry

16

Interface Flexibility

Only Thermo has made progress in this

critical ICP-MS aspect in the last 10

years!

Default Xt Interface

• Analyses heavy matrix samples without

drift

Optional Xs Interface

• Xs-

• For applications requiring higher sensitivity –

especially at higher masses

• Laser, LC, GC etc…

• Xs+

• New in the XSERIES 2

• Protective Ion Extraction

Ultimate BECs on low mass elements

Materials, semicon…

17

NEW TOOLS: ESI – FAST

18

NEW TOOLS: ESI – FAST

Increase of throughput

Decreasing of contamination

due to memory effects from

peristaltic pump tubing

On-line Internal standard

addition

19

ICP-MS: high-end for elemental analysis

It can measure almost the whole periodic table in just about everything

It is not limited to elemental concentrations but offers high precision

isotope ratio determinations and, if coupled to separation devices,

species information

20

Elements or species ?

HO-As-OH

|

OH

O

||

H3C-As-OH

|

OH

O

||

H3C-As-CH3

|

OH

CH3

|

H3C-As+-CH2CO2-

|

CH3

CH3

|

H3C-As+-CH2CH2OH

|

CH3 Br-

BrO-

21

Definition - „the analytical activity of identifying and/or measuring the

quantities of one or more individual chemical species in a sample‟

IUPAC

That means – chemical species of an element are determined rather

than the total element concentration

Example – Tin species separated by gas chromatography (GC) and

detected by ICP-MS

What is Speciation Analysis?

Sn Sn

Sn

22

Why do we need Speciation…?

Bioavailability

Environmental Fate

Mobility

Toxicity

Volatile Elemental

Species

Multi-Element/Isotope

High Sensitivity

High Selectivity

Process Control

Chemistry

Sn Sn

Sn

23

The chemical form can be very important…….

24

Elemental speciation data can reveal valuable information in addition to total element concentrations:

Speciation in Environmental, Food and Clinical Analysis

Environmental Fate

Mobility

Metabolic Processes

Bioavailability/toxicity

Reactivity

Industrial

Sources

Natural

SourcesHg0

Hg2+ MeHg+

25

Growth of Speciation

26

HPLC-ICP-MS Applications

Element Species Matrices Scientific Sector

Cr Chromium III & VI Water, air particulates, cementEnvironment, industry, nutrition,

occ. exp.

As Organoarsenic compounds Fish, water, sediments, urine, hair Environment, nutrition, exposure

SeOrganoselenium Yeast, garlic, urine Nutrition, pharma, clinical

Hg Inorganic mercury, organomercury Fish, sediments, water, urine, blood Environment, Clinical

Sn Organotins Fish, sediments, water, plastics Environment, industry

Fe Iron II & III Bacteria, soils, water Environment, clinical, nutrition

Cu, Zn, Cd Phytochelatins, metallothioneins Plant tissues, brain & kidney tissue Agroindustry, clinical

27

Speciation applications

Polypeptide sub-isoforms

Metallodrugs hydrolysis products

Metalloporphyrins and corrinoids

Organoselenium

Organoarsenic

Size-exclusion

Reversed-phaseAnion-exchage

Cation-exchageIon-pairing

Polysaccarides

Metallothioneine

Phytochelatins

Metalloenzymes

Transport proteins

Metallodrug-protein adducts

Polypeptide sub-isoforms

Redox states

Organoselenium

OrganoarsenicHPLC-ICP-MS

28

CHEMICAL FORM LD50 (mg/kg)

Arsenite (As(III)) 14

Arsenate (As(V)) 20

Arsine (AsH3) 3

Monomethylarsonic acid (MMA) 700 - 1800

Dimethylarsinic acid (DMA) 700 - 2600

Arsenocholine > 10000

Arsenobetaine > 10000

Aspirin 1000 - 1600

Strychnine 16

LD50 rat: Lethal dose 50 (dose for which 50 % of a population is dead)

Arsenic Speciation – Why?

The toxicity of arsenic is strongly dependent on the chemical

form

29

Arsenic Speciation – Why?

Arsenic is accumulated by marine biota

• Fish

• Shellfish

• Algae

Marine biota are able to transform arsenic species, creating

many arsenic containing species (over 30 As species have been

identified to date)

Upon consumption of marine biota, toxicity risk assessment is

needed and detoxification pathways can be elucidated

Presence of arsenic in fish and marine based products has also

disrupted international trade

30

Speciation of As by Anion Exchange HPLC-ICP-MS

Optimization of HPLC parameters

Surveyor LC

ANION EXCHANGE: Hamilton PRP x-100, 250 x

4.6 mm, 5μm

• Gradient elution with 20-50 mM ammonium

carbonate buffer, pH 8.9

• Flow rate: 1mL/min, Inj. vol: 20 μL

1 ppb of each As standard

Check:Simple coupling

Plasma robustness

Organic solvents

Interference strategy

Data evaluation

31

Speciation of As in Seafood by Anion Exchange HPLC-ICP-MS

Sample Prep Protocol

Fish samples – human consumption

• Filets sliced into small cubes

• Freeze-dried to remove moisture

• Ground to a fine powder with pestle and mortar

Extraction protocol

• 250mg fish sample extracted with 10mL H2O in ultrasonic bath at approx. 60°C

32

As Speciation in Commercially Available Fish

Arsenic Containing Species (µg As/g) in Dried Fish

CRM Source AsB AsIII DMA

Unknown

A MMA AsV

DORM-2 (Dogfish

muscle) IRMM, Belgium 15,6 ± 1,04 - 0,2 ± 0,03 - - -

Sample Source

Red tuna North-East Atlantic 3.9 - 0.11 - - 0.13

Canned tuna Ecuador 3.42 - 0.12 - 0.08 -

Fillet of Trout Farmed, France 3.68 - - - - -

Freshwater Perch France 0.06 - - - - -

Ling North-East Atlantic 45.35 - - - - -

Alaskan Hake fillets* Pacific Ocean 12.78 - - - - -

Squid Heads* Indian Ocean 1.93 - - 0.02 - -

Arsenic Containing Species Recovery (%)

Sample Source AsB AsIII DMA

Unknown

A MMA AsV

Canned tuna Ecuador 97.6 115.8 104.8 - 114.1 129.1

Squid Heads* Indian Ocean 118.3 112.4 106.5 - 114.2 123.7

33

Speciation of As in Urine by Anion Exchange HPLC-ICP-MS

Sample Y: Non-fish eater, non-occupationally exposed individual (smoker)

Sample X: Fish eater, non-occupationally exposed individual (non smoker)

34

Mercury Speciation – Why?

Industrial

Sources

Natural

SourcesVegetation

Atmospheric

Dispersion

Ocean

Natural sources:

• Volcanic activity

• Geothermal activity

Anthropogenic sources:

• Bactericide

• Manufacturing processes

• Fossil fuel combustion

Daily exposure:

• Fish consumption (MeHg+)

• Dental amalgams (Hg0)

• Vaccines – ‘Thimerosal’

Occupational exposure:

• Cinnabar (HgS) purification

• Gold mining

• Industrial processes

35

Mercury Speciation - Toxicity

Toxicity of Hg is species dependant• Organomercurials are more easily absorbed and less readily eliminated than

inorganic Hg

Mobility and reactivity

• highly soluble in lipids

• transfers easily into blood

• crosses cellular membranes, blood-brain barrier

• pro-oxidant

• high affinity for free sulfhydryl groups and Se

Neurotoxin

• impairs mental and neurological function

• sensory and motor deficits

• also compromises hormone metabolism and inhibits protein and RNA synthesisGroups at risk:

Pregnant, nursing women, women of childbearing age and young children are in the highest group at risk because of the high sensitivity of foetus and child brain in development

36

Mercury Speciation for Risk Assessment

Provisional Tolerable Weekly Intakes

• Set by international scientific committees based on

threshold values for the effects on the

neurodevelopment of children

• 0.7 µg/kg body weight/week (NRC, 2000)

• 1.6 µg/kg body weight/week (JECFA, 2003)

International fish consumption

recommendations:

• EPA – pregnant women should limit consumption of

marine food to 6 ounces/week

• FDA – pregnant women should avoid consumption

of shark and swordfish

• EFSA – pregnant women should limit consumption

to 2 portions of fish/week, avoid swordfish and tuna

Amplification factor in food

chain means larger

predatory fish contain

higher levels of MeHg+

37

High Mercury Levels Are Found in Tuna Sushi

“Tuna sushi is a popular item

in New York but may be risky.”

By MARIAN BURROS

Published: January 23, 2008

“Recent laboratory tests found so much mercury in tuna

sushi from 20 Manhattan stores and restaurants that at

most of them, a regular diet of six pieces a week would

exceed the levels considered acceptable by the

Environmental Protection Agency. “

38

Speciation of Mercury by RP HPLC-ICP-MS

HPLC column: Hypersil GOLD

(150 x 4.6 mm, 5µm)

Isocratic elution:

60 mM ammonium acetate, 5%

methanol, 0.01% 2-

mercaptoethanol

Flow rate: 1.5 mL/min

Inj. vol: 100 μL

HPLC parametersRun time 10 min

Sample

throughput

120/day

BEC 0.007 ng/g

LOD 0.05 ng/g

39

Speciation of Hg in Fish (DORM-2) and Urine

Urine (plus spike)

DORM-2 (plus spike)

Extraction:

Analytical methodology (3 min.)

+ 5 mL TMAH

Microwave 2 min 40W

Fish sample prep

Urine sample prep

Analytical methodology (0 min.)

Method validation:

[MeHg+] = 4.27 ± 0.12 μg/g

(96% of certified value)

MeHg+

Hg2+

MeHg+

Hg2+

40

Thermo Scientific ICP-MS Speciation Packages

41

ELEMENT2/XSERIES 2 LC Coupling

42

ICP-MS for GC applications

Unique DUAL MODE sample introduction

• Three legged GC-ICP-MS torch

• Axial introduction of GC gas

• Aspirated solution introduced via 2nd leg

• GC transferline slots into holder in Faraday cage and is clamped into place

• Spray chamber is mounted via a specially configured adapter above the GC transfer line

43

Recent Development of GC Interface for ELEMENT 2

44

Advantages

Easy setup, tuning and performance testing with aqueous solution

Gas and solution analyses are possible without reconfiguring the

interface

Robust plasma conditions for GC-ICP-MS analysis due to wet plasma

On-line addition of aqueous internal standards

Entrained air from spray chamber burns up carbon of the solvent GC

injection, preventing carbon build up on the cones and improving long

term stability (there is no need for additional oxygen)

Sensitive multi-element capability

Speciation and total element analysis

•

45

GC-ICP-MS Applications

Elements Elemental Species Matrices Scientific Sector

Sn Organotins (TBT) Sediments, fish, water, plastics

Environment, nutrition, manufacturing,

packaging

Hg Organomercury (MeHg+)

Fish, sediments, water, blood,

petrochemicals

Environment, nutrition, exposure,

industry

SThiophene analogs and Hg

compounds Petrochemicals Petro Industry, environment

Br Flame retardants (PBDEs) Household dust, blood, milk, fat Clinical, industry, environment

Cl, P, S Pesticides Vegetables, fruit Agroindustry, environment

Pb Organolead Sediments, urban dust Environment

I Iodine species Thyroid, milk, seaweed Nutrition, clinical, environment

S, As, Sb, Ge,

Ga, Th,.. Organometallic gases Gas products Semiconductor industry, gas suppliers

C VOC Gas Industry, environment

46

Speciation of Mercury by GC-ICP-MS

Column Thermo Fisher Scientific Tr-5, 30 m x0.25mm ID, 25 μm

Injection mode PTV, splitless

Injection porttemperature

250 C with ramp to 400 C

Injection volume 1 µL

Carrier gas flow He @ 3 mL min-1

GC ovenparameters

50 C (1 min), ramp at 30 C/min to 300 C (1min)

Nebulizer andspray chamber

Concentric with impact bead (freeaspiration)

Nebulizer gas 0.35 L min-1

Additional gas Ar @ 600 mL min-1

Forward power 1400 W

47

GC-ICP-MS Speciation of Hg in Blood

Blank

NIST bovine blood

966 SRM

48

Speciation of Butlyltins – Why?

Sources of organo-Sn to the environment:

• PVC stabilisers ,fungicides, biocides, rodent repellents, wood preservatives, anti-foulants on ships and quays

Persistent compounds, resistant to degradation in water

Bio accumulate readily in fatty tissues

Broadly classified as endocrine disrupters & toxic to aquatic organisms at nano-molar concentrations

1970 1980 1990 2000

Occurrence of organotins in

open ocean waters and

accumulation in sea mammals

Ev

olu

tion

of

org

anot

inco

mp

ound

s co

nce

ntr

atio

ns in

co

asta

lwat

ers

(arb

itrar

yu

nits

PP

T)

Bio

log

ical

impa

cts

of

org

ano

tinco

mpo

unds

Analytical information

Increasing sensitivity and selectivity

CT/QFAAS

CGC/FPD or AEDCGC/ICP-MS

Time

Occurrence of organotins

in fresh water systems

and in household products

GFAAS

Oyster toxicity

and biological impacts in

coastal environments

Imposex phenomenom

in coastal and open seawater

ways

Ban of the use of TBT on

small vessel less than 25

meters

SnSn

Sn

49

Speciation of Butlyltins by GC-ICP-MS

Column Tr-5, 30 m x 0.25 mm i.d., df 0.25 µm

Injection mode Splitless

Injection port temperature 280 C

Injection volume 1 µL

Carrier gas flow He @ 1.5 mL min-1

Make up gas flow* Ar @ 400 mL min-1

Transfer line temperature 300 C isothermal

Initial temperature 60 C for 1 min

Ramp rate 50 C min-1

Final temperature 280 C for 1 min

Species LOD pg/g

MBT 3 (4 fg)

DBT 3 (4 fg)

TBT 1.7 (2.4 fg)

50

Conclusions

Flexible speciation packages for HPLC and GC applications

• Bi-directional communication in both speciation packages for automated speciation analysis

• Transient data processing integrated

• Unique dual mode sample introduction system for GC-ICP-MS

• Robust plasmas

Many chromatographic methodologies already developed and validated

Speciation is matrix dependent

• A method developed for fish may not necessarily work for blood, due to matrix components which

can disrupt extraction strategies or chromatographic elution

• The sample preparation is a major step in speciation analysis; the maximum of species should be

extracted from the tissue without changing the chemical form in the process

Low limits of detections (ppt range) are routinely achieved

• GC-ICP-MS tends to allow lower LODs but is not as flexible as HPLC

• Sector-Field ICP-MS delivers lower LODs compared to Quadropole ICP-MS

Several Legislations exist:

51

Speciation and Legislation

Element Matrix International Agency/Directive Legislation

Hg

Fish USEPA Criterion document 823-R-01-001 Criterion limit value of 0.3 mg/kg of MeHg+ (concentration of

MeHg+ in fish which is expected to be without appreciable risk to human health)

Water EU Water Framework Directive 2000/60/ECHg and its compounds listed as a priority hazardous

substances

Sn Water EU Water Framework Directive 2000/60/EC TBT listed as a priority hazardous substance

Cr

Automobiles EC directive 2000/53/EC Maximum of 2 g of Cr(VI) per end-of-life vehicle

Cement EC directive 2003/53/ECMaximum of 2 mg/kg Cr(VI) in cement or cement

preparations which will come into contact with skin

Workplace

AtmospheresUSEPA and ACGIH

Threshold Limit value of 0,05 mg soluble Cr(VI)/m-3 air and

0,01 mg insoluble Cr(VI)/m-3 air

Br Water EU Water Framework Directive 2000/60/ECPentabromodiphenyl ether listed as a priority hazardous

substance

Many governing bodies are now recognizing that the elemental species and not necessarily the total concentration of an element should be controlled

Legislation for elemental species focuses on the more toxic species

Future amendments to the Water Framework Directive in the EU will require maximal concentrations of 50 ppt for Hg compounds and 0.2 ppt for Tributyl tin in inland water systems

52

Traceability - REE

53

Tomato sauce – different countries

-0,01

0

0,01

0,02

0,03

0,04

0,05

0,06

µg

/g

Ce Dy Er Eu Gd Ho La Lu Nd Pr Sm Tb Tm Yb

# 1 # 2 # 3 # 4 # 5 # 6 # 7 # 8

54

Tomato sauce – REE analysis RRE of #1 vs. [# 2-8] average

0

0,01

0,02

0,03

0,04

0,05

0,06

µg

/g

Ce

Dy

* 10

Er*1

0

Eu

*10

Gd

Ho

*10

La

Lu

*10

Nd

Pr

Sm

Tb

*10

Tm

*10

Yb

# 1 Media # 2-8

55

LOD

139La

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175Lu

0.000

0.020

0.040

0.060

0.080

0.100

0.120

0.140

0.160

pp

tDETECTION LIMITS

LOD: < 0,2 ng/L

56

repeatability

0

5

10

15

20

139L

a

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175L

u

RSD % (10 replicates)

10 ppt 1 ppt 0,5 ppt

57

Forensic case: water contamination

0

5

10

15

20

139L

a

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175L

u

Home driver sample #1

0

10

20

30

40

50

139L

a

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175L

u

refinery plant

0

5

10

15

20

25

139L

a

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175L

u

tank truck

0

2

4

6

8

10

12

14

16

139L

a

140C

e

141P

r

146N

d

147S

m

153E

u

157G

d

159T

b

163D

y

165H

o

166E

r

169T

m

172Y

b

175L

uHome driver sample #2

58

APPLICAZIONI III: TRACCIABILITA’

139L

a

141P

r

147S

m

157G

d

163D

y

166E

r

172Y

b

S.ANNA

LAURA

S. FRANCESCOTAVINA

S. PELLEGRINOS. BERNARDO

LAURETANAGAUDIANELLO

0

10

20

30

40

50

60

70

80

PATTERN REE

S.ANNA LAURA S. FRANCESCO TAVINA

S. PELLEGRINO S. BERNARDO LAURETANA GAUDIANELLO