Nuevas fronteras en

Investigación con el

nuevo ICP-MS/MS

Agilent 8800

Transformando la

tecnología en ICP-MS

1 March 20, 2014

Rubén García Especialista de Producto, Agilent Technologies

New 8800 ICP-MS/MS - Outline

1. Introduction to the new 8800 ICP Triple Quad

2. Key technology in the 8800

3. Unique modes of operation with MS/MS

4. 8800 performance in some key applications

2 March 20, 2014

Agilent 8800 ICP-MS/MS

World’s first Triple Quadrupole ICP-MS (tandem MS)

New modes of operation and performance not possible with quadrupole ICP-MS

Joins the Agilent 7700, Agilent’s high performance quadrupole ICP-MS system

Unique capabilities, based on proven technology (shared sample introduction, HMI,

many consumables, robustness, key hardware components and software platform)

Agilent 7700

Single-quad

(ICP-QMS)

New Agilent 8800

ICP-MS/MS

3

Why ICP-MS/MS?

March 20, 2014

New Agilent 7900

Single-quad (ICP-

QMS)

Interference Removal in Quadrupole ICP-MS

7700 Series with He mode is well-known for accurate multi-element analysis of

unknown, variable and high-matrix samples (enviro, food, clinical, pharma…)

He mode on 7700 is effective for ALL polyatomic interferences at low- and sub-ppb levels

BUT He mode can’t remove/avoid isobaric overlaps (e.g. 40Ar on 40Ca) or doubly-

charged interferences (e.g. 150Sm++ and 150Nd++ on 75As+). Reaction mode can…

Also, reactive gases can give more complete removal of interferences when ppt

level detection is needed (H2 and NH3 commonly used in semicon applications)

4

Complex matrix in no gas (above) and

He mode (right, with standard inset)

7700 removes ALL polyatomics

7700 - He mode for Polyatomic Interferences

No gas mode (left) and He mode (below)

March 20, 2014

Reaction Gases for ICP-QMS and ICP-MS/MS

Limitations of reactive cell gases in quadrupole ICP-MS are well-

documented:

All ions enter the cell, affecting reaction processes. Gives variable results when

sample type/matrix or co-existing analytes change

Product ions from matrix or other elements can create new overlaps on analytes

Analyte product ions can be overlapped by other analytes/matrix elements

Tandem MS configuration eliminates the variability caused by co-existing

elements and changing matrix components, as Q1 rejects all masses

except the target analyte and on-mass interferences

Results are consistent and reliable

5

Agilent 8800 ICP-MS/MS matches 7700

performance in He mode, but also

offers controlled and consistent MS/MS

operation in reaction mode.

March 20, 2014

8800 ICP-QQQ Key Product Hardware

6 March 20, 2014

Low flow

sample

introduction

system

High matrix

introduction

(HMI)

technology

Fast, frequency-

matching 27MHz

RF generator Efficient twin-turbo

vacuum system

Dual conical Extraction and

Omega lens focuses ions

across the mass range

9 orders

dynamic range

electron

multiplier (EM)

detector

Analyzer quad Q2:

High frequency

hyperbolic quadrupole

– selects ions that

pass to detector

High-transmission,

matrix tolerant interface

First quad Q1: High frequency

hyperbolic quadrupole mass filter –

selects ions that enter the cell

3rd generation collision/

reaction cell (ORS3)

with 4 cell gas lines

Peltier-

cooled

spray

chamber

Robust, high-temperature

plasma ion source

Introduction to INORGANIC MS/MS

This is the very first and only instrument to make MS/MS

technology available to inorganic analytical chemistry

Completely novel

Allows the determination of elements that would otherwise be

“difficult” for quadrupole (and SF) ICP-MS

Elements such as Sulphur and Phosphorus become “laughably”

easy!

Uniquely, Isotopic information is retained when using reaction

mode

March 20, 2014 7

QQQ in Organic MS

170 210 250 290

210

222

268 280 165

Quad Mass Filter (Q3) Quad Mass Filter (Q1)

Collision Cell

Spectrum with

background

ions (from ESI)

Q1 lets only

target ion 210

pass through

190 210

210

Collision cell

breaks ion 210

apart

150 170 190 210

210 158

191

Q3 monitors only

characteristic

fragments 158

and 191 from ion

210 for quant and

qual.

160

158

190

191

no chemical background

QQQ in Inorganic

Unlike Organic QQQ we do not split the parent ion into product

ions

Not trying to split the atom!

March 20, 2014 9

ICP-MS/MS: How Does it Work?

ICP (plasma) and Interface: Forms and

extracts ions from the sample (just like the

7700 or 7900)

EM (detector): Measures the

ions that are scanned by Q2

(just like the 7700)

10

Q1 – controls ions

that enter the cell

• Consistent reactions

even if sample

composition changes

ORS3 – collision/

reaction gas added

• Ions react and are

neutralized or moved

• Product ions are formed

Q2 – selects the

target analyte mass

• Interference-free analyte

ions passed to EM

March 20, 2014

Q1 – controls ions

that enter the cell

• Consistent reactions

even if sample

composition changes

ORS3 – collision/

reaction gas added

• Ions react and are

neutralized or moved

• Product ions are formed

Q2 – selects the

target analyte mass

• Interference-free analyte

ions passed to EM

ICP-MS/MS: How Does it Work?

ICP (plasma) and Interface:

Forms and extracts ions from

the sample (just like the 7700)

EM (detector): Measures the

ions that are scanned by Q2

(just like the 7700)

Unique aspect of 8800 is MS/MS Mode

• Q1 controls the ions that enter the cell

With MS/MS, reaction processes in the cell

are consistent and predictable, even if

sample composition is complex or variable

• Consistent cell conditions means reaction

mode method development is much

easier than on single quad (same method

can be used for different samples)

11 March 20, 2014

Operational Modes

• Q1 opens allowing all ions into ORS3

Single Quad

• Q1 allows a range of m/ƶ into ORS3 Single Quad with Band Pass Filter

• Both Q1 and Q2 set to same mass MS/MS – On

Mass

• Q1 and Q2 set to different masses MS/MS – Mass

Shift

Unique to the 8800

Quad Cell m/z

Ions

Reaction Gas

M0

Single Quad Configuration Analyte (unreactive) Interference (reactive)

M1

If the ions of mass M1 react with the gas to form ions of mass M0, they act as new

interference.

ICP-QQQ-MS Operating Principles

Q2 Cell

Ions

Q1

(set to M0)

Reaction Gas

m/z of Q2

M0

QQQ Configuration

M0

M1

Analyte (unreactive) Interference (reactive)

ICP-QQQ-MS Operating Principles

Quad Cell

Ions

Reaction Gas

M0

Single Quad Configuration

-Mass Shift Method- Analyte (reactive) Interference

M2

If there are unreactive ions of M2, the mass shift method does not work.

ICP-QQQ-MS Operating Principles

M0

QQQ Configuration

-Mass Shift Method- Analyte (reactive) Interference

M2

Q2 Cell

Ions

Q1

(set to M0)

Reaction Gas

M0

m/z of Q2

ICP-QQQ-MS Operating Principles

Which Applications Need ICP-MS/MS?

Applications that may benefit from ICP-MS/MS with Agilent 8800.

• Environmental: As and Se analysis in presence of REE. REE++ overlap As+ & Se+.

• High purity chemical: Ti and Zn analysis in semiconductor grade H2SO4 / H3PO4. S and P

polyatomic ions such as SO+, PO+ and SO2+ overlap Ti+ and Zn+.

• Material: P in Si matrix. SiH+ and SiH2+ overlap on P+.

• Material: As in Co matrix. Fe and Ni in Ca matrix. MoO+/MoOH+ interference on Cd.

• Geology: REE analysis. BaO and REE-O ion overlap other REE.

• Food: Sulfur Isotope Ratio analysis.

• Clinical: Ti and Cr analysis in blood and serum. S , P and C matrix.

• Nuclear: 129Iodine analysis. 129Xe atomic isobar interference.

• Nuclear: Long lived radio nuclide analysis. 93Zr, 99Tc, 135Cs and so on.

• Life Science: Trace Sulfur for protein/peptide quantification.

• …

17

Mercury Interference on Lead

18

Pb measurement - Mercury Reaction with Ammonia Detailed version presented at Goldschmidt Conference

Mercury undergoes a charge-transfer reaction:

Hg+ + NH3 Hg0 + NH3+

(NH3+ + NH3 NH4

+ + “NH2”)

This reaction is very rapid and essentially 100% efficient

This means that 204Hg could be eliminated as an overlap on 204Pb if Pb does not react in the same way

This is an important measurement in geochronological

measurements or isotope tracer studies (e.g. spike a specific

isotope and trace protein interaction)

Pb, Tl at 1ppb in presence of 10ppb Hg

21

No Cell Gas

Hg

interference

on 204Pb

isotope

Pb Isotopes

do not match

theoretical

template

Impossible to

measure the

natural Pb

contribution

204Pb Clearly overlapped by 204Hg

Pb, Tl at 1ppb in presence of 10ppb Hg + 100ppb REE’s

25

NH3 Cell

Gas NOT

USING

MS/MS

All isotope

patterns a

complete

mess from

REE cluster

ions

Mass Cluster

204 (Pb) Eu(NH3)3

Yb(NH3)2

Ce(NH3)4

205 (Tl) Yb(NH3)2

Gd(NH3)3

206 (Pb) Yb(NH3)2

Lu(NH2)2

La(NH3)4

Ce(NH3)4

Gd(NH3)3

207 (Pb) La(NH3)4

Yb(NH3)2

Gd(NH3)3

208 (Pb) Ce(NH3)4

GdNH(NH3)2

TbNH(NH3)2

Yb(NH3)2

Gd(NH3)3

Pb, Tl at 1ppb in presence of 10ppb Hg + 100ppb REE’s

26

MS/MS NH3

Cell Gas

All isotope

patterns

match theory

again.

MS/MS

controls the

reaction

chemistry

Pb, Tl at 1ppb in presence of 10ppb Hg + 100ppb REE’s

27

MS/MS NH3

Cell Gas

All isotope

patterns

match theory

again.

MS/MS

controls the

reaction

chemistry

As 204Hg isotope is more abundant than 204Pb; “real” interference is ~5x higher

MS/MS – On Mass Measurement - Pb

All none on mass interferences rejected by Q1

Mercury-based interferences react with ammonia through charge transfer and are neutralised. The neutral atom is pumped to “waste” in vacuum chamber

28

< 204 >Hg+

< 204 >Pb+

All REE’s+

204Hg+

204Pb+

< 204 >Hg+

< 204 >Pb+

All REE’s+

204Hg0

204Pb+

Reaction gas

NH3

1st Quad (Q1) Rejects ALL masses except

analyte (204Pb) and on-mass

interferences (204Hg+

ORS3 Cell Converts Hg+

to Hg0

Product

2nd Quad (Q2) Set at Q1

Rejects all cell

formed ions and

neutral species from m/z 204

Tungsten Interference on Mercury

29

MS/MS – On Mass Measurement - Hg

All none on mass Hg and W-based interferences rejected by Q1

Tungsten-based interferences react with oxygen and are mass shifted away from selected Hg isotopes

30

195 196 197 198 199 200 201 202 203 204

195 196 197 198 199 200 201 202 203 204

MS/MS for Hg with O2 Cell Gas

MS/MS On-Mass Mode – separation of Hg+ from WO+ overlap

Spectrum of 2ppb Hg (top) and 5ppm

W (bottom) on the same scale.

WO+/WOH+ overlap is reduced to

<10cps at 201Hg

31

Molybdenum Interference on Cadmium

32

Molybdenum Oxide Interference

Molybdenum is a fairly prevalent element within the environment

• The 54th most abundant element in the Earth's crust and the 25th most abundant element in the oceans

It is an alloying agent in many steels

It is an important cofactor and constituent of over 50 enzymes

• An important micronutrient

Cadmium is toxic and (usually) present at low concentration

It is important to be able to reliably measure Cd at low concentrations in the presence of Mo

Mo has several isotopes, all forming interferences on Cd

SF ICP-MS requires resolution of >32,000

MoO Interference

34

No Cell Gas

1mg/Kg Mo

MoO

interference

on all

available Cd

isotopes

MoO Interference

35

No Cell Gas

1mg/Kg Mo

+ 1ug/Kg Cd

MoO

interference

on all

available Cd

isotopes

Peak pattern

does not fit

MoO Interference

36

MS/MS NH3

Cell Gas

1mg/Kg Mo

+ 1ug/Kg Cd

MoO

interference

completely

removed

Excellent fit

for all Cd

isotopes

1,028

1,119

1,565

2,075

1,097

0,991 0,992 1,003 0,99

0,0005 0

0,5

1

1,5

2

2,5

20ppb Mo + 1ppb Cd 100ppb Mo + 1ppb Cd 500ppb Mo + 1ppb Cd 1000ppb Mo + 1ppb Cd 1000ppb Mo

No Gas

MS/MS NH3

Cadmium: Recovery in Variable Mo Matrix

37

Special Alloy Test, Trace Mg with Ti

38

Ti++ Interference on Mg in Specialist Alloy

Special Ti-based alloy – Ti concentration ~0.5g l-1 in Aqua

Regia; Mg very low (<1ppb in solution) but Ti++ biases data

39

No Cell Gas Ammonia Cell Gas

Isobaric interference removal: I-129

Rest of ions

including I-127 Removal of Xe

O2 as reaction gas and MS/MS- on mass

Reaction

gas (O2)

Xe129

I129

I+

Xe

Isobaric interference removal: I-129

O2 completely removes

the Xe and now the I-129

can be detected at

ultratrace levels

Without interference removal

, the isotopic pattern of Xe

interferres I-219 (and I-127)

Isobaric interference removal: I-129

Detection limit obtained using the calibration curve:

0.025ppt.

Very low Blank Equivalent Concentration: 0.25ppt

due to the complete removal of the Xe-129

Calibration

curve using the

I-129 standard

Ejemplo de MS/MS en modo “on-mass” con N2O en la celda

45 March 20, 2014

Eliminación de interferencias isobáricas y poliatómicas en Selenio

Se+ + N2O apenas reacciona

Br+ + N2O BrO+ (totalmente)

Q1 – elimina todas las

masas excepto la del analito

(Se+ y su interferencia Br+)

ORS3 – se forma el BrO+

Q2 – Elimina todos los iones

formados en la celda distintos al

ion analito Se+, dejando a este

libre de solapamientos

Gas de Reaccion (N2O en He)

79Se

79Br

Se+

BrO+

Adicionalmente las interferencias ArAr, BrH son eliminadas por efecto colisional con

Helio

Determinación de Se en modo reacción con N2O

March 20, 2014

Confidentiality Label

46

1 ppb Se: Sin interfererencia ArAr o BrH

Desplazamiento 79Br y 91Br a masas

95 y 97

Datos cortesía ISC-Science y

Universidad de Oviedo

Operational Modes

• Q1 opens allowing all ions into ORS3

Single Quad

• Q1 allows a range of m/ƶ into ORS3 Single Quad with Band Pass Filter

• Both Q1 and Q2 set to same mass MS/MS – On

Mass

• Q1 and Q2 set to different masses MS/MS – Mass

Shift

Operational Modes

• Q1 opens allowing all ions into ORS3

Single Quad

• Q1 allows a range of m/ƶ into ORS3 Single Quad with Band Pass Filter

• Both Q1 and Q2 set to same mass MS/MS – On

Mass

• Q1 and Q2 set to different masses MS/MS – Mass

Shift

48

High Purity Materials.

Trace elements in HCl

Ti and V in 10% H2SO4

P and Ti in 2000ppm Si

49

Ge and As Measured as GeO+ and AsO+ in 20% HCl DLs 1.5ppt for Ge and 2.6ppt for As

MS/MS – Removal/avoidance of Cl-based interferences on 74Ge and 75As

In 20% HCl, Ge and As are key interfered elements (Cl2+, ArCl+).

• Both analytes react quickly with O2 cell gas and can be measured at low levels as

their oxide product ions.

50 March 20, 2014

V and Ti Measured in 9.8% H2SO4 (1:10 dilution)

In 10x diluted (9.8%) H2SO4, V and Ti are key interfered elements (SO+, SOH+).

• V reacts slowly with NH3 cell gas, so 51V+ can be measured on-mass.

• Ti reacts quickly with NH3, so 63TiNH+ (and other) product ions can be measured.

MS/MS on-mass and mass-shift can be combined in the same acquisition method.

DLs and BECs for both elements are single-ppt or sub-ppt

51V measured on-mass 48Ti measured as 63TiNH+

51 March 20, 2014

P Measured as PH3+

in 2000ppm Si/HF

H2 cell gas mode - DL 170ppt

C-flow 50 nebulizer P is a critical

contaminant in solar Si

(for PV cells)

Difficult to measure at

low levels in high silicon

matrix due to

interference from 30SiH

8800 with MS/MS allows

controlled reaction mode

with H2 cell gas to

measure P at low levels

as its reaction product

ion PH3+ at mass 34

52 March 20, 2014

48Ti Measured as TiNH+ in 2000ppm Si/HF

NH3 cell gas mode - DL 6.2ppt

C-flow 50 nebulizer Ti is very difficult to

measure at low levels in

high Si matrix (29Si19F

and 28Si19FH overlaps

on the major Ti isotope

at mass 48)

8800 MS/MS in NH3 cell

gas mode moves Ti

away from matrix-based

overlaps and allows low

ppt level measurement

of Ti as TiNH+ product

ion at m/z 63

53 March 20, 2014

Sulphur, Arsenic and Selenium

54

Sulphur is difficult!

Interferences from Oxygen, Nitrogen and Hydrogen polyatomics:

32S: 16O16O; 14N18O; 15N16O1H

33S: 32S1H; 16O16O1H; 16O17O; 15N18O; 14N18O1H

34S: 33S1H; 33S1H2; 16O18O

…and more!

55

Sulphur Measurement

With conventional Collision-Reaction Cell (CRC) ICP-MS interferences can be reduced but not necessarily to a level that is useful or for all isotopes of interest.

Due to the intensity of the interferences a mass-shift (neutral gain) methodology is often employed.

• S measured as SO at +16amu

– If another isotope resides at the +16 mass (e.g. 48Ca & 48Ti on32S16O) analysis will be impaired

– All S isotopes react with all O2 isotopes causing low mass S isotopes to interfere with higher mass isotopes

• 32S17O+ interference on 33S16O+

• 32S18O+ & 33S17O+ interference on 34S16O+

56

Sulphur (O2 Mode) - Summary

Most S isotopes can be measured using 16O mass

shift:

• 32S – SO+ @ 48

• 33S – SO+ @ 49

• 34S – SO+ @ 50

Limiting factor for the analysis of S is now the

background signal due to contamination.

S is commonly present as a contaminant in reagents,

plastic-ware (e.g. phthalates)

57

Oxygen Mass Shift for 34S – with MS/MS

58

50Cr+/50V+/50Ti+

38Ar12C+

13C37Cl+

34S+

17O2+

16O18O+

O2 reaction gas

34S16O+

Q1

MS/MS

34amu

Q2

50amu

The mass difference between Q1 and Q2 is fixed (16) therefore a single

transition is observed – the other oxygen isotopes are eliminated so the

original isotopic pattern is preserved!

59

ICP-MS/MS: Mass Shift with O2 Cell Gas

32S16O+

33S16O+

34S16O+

34S/32S ratios

Blank 0.0484 ± 0.0017

100 ppb standard 0.0487 ± 0.0012

Theoretical 0.0447 ± 0.0025

0

10.000

20.000

30.000

40.000

50.000

60.000

47 48 49 50 51 52 53

ion

co

un

ts [

cp

s]

m/z of Q2

32S16O

33S16O

34S16O

36S16O 36Ar16O

0

2.000

4.000

6.000

8.000

10.000

47 48 49 50 51 52 53

ion

co

un

ts [

cp

s]

m/z of Q2

Sulphur in DIW and AcN – limit is contamination

0

1.000

2.000

3.000

4.000

5.000

47 48 49 50 51 52 53

ion

co

un

ts [c

ps

] m/z of Q2

5ppb S in UPW UPW only AcN

60 March 20, 2014

Publication on Sulphur by Isotope Dilution

61

REE++ Interferences on As and Se

M++ interferences are not polyatomic, so not removed by He cell gas

In He mode, REE++ interferences can actually appear to be higher due to

transmission efficiency of heavier (and slightly smaller) 2+ ions

• Nd & Sm have isotopes at m/z 150, so the 2+ ions appear at m/z 75 (As)

• Eu has an isotope at m/z 151, so the 2+ ion appears at m/z 75.5 (As)

• Gd has an isotope at m/z 156, so the 2+ ion appears at m/z 78 (Se)

Sample 75 [ No Gas ] 75 [ He ] 78 [ No Gas ] 78 [ He ] 75 -> 91 [ O2 ] 78 -> 94 [ O2 ] La 500ppb 0.000 0.000 0.000 0.559 0.000 0.004

Ce 0.000 0.000 0.000 0.109 0.000 0.001

Nd 0.730 5.890 0.000 0.000 0.000 0.000

Sm 0.634 8.916 0.000 0.000 0.002 0.001

Eu 1.761 16.321 0.000 0.000 0.000 0.001

Gd 0.000 0.000 34.728 290.035 0.000 0.007

Tb 0.000 0.000 0.000 0.000 0.000 0.000

Dy 0.000 0.000 0.000 0.423 0.000 0.000

62 March 20, 2014

O2 cell gas moves As and Se to AsO+ & SeO+ product ions, so avoiding the REE++

overlaps

Some Solutions in Clinical Sample Analysis

63

Gd++ Interference on Selenium

Gd used as a contrasting

agent for MRI

Concentration in patient can be at the high ppb to

ppm level

This causes problems with

obtaining accurate Se

measurements for those patients

Using oxygen to shift Se mass

+16amu effectively

removes bias

No Gas Oxygen

Serum 93.64 91.42

Serum + Gd 250 µg l-1

99.87 91.38

Serum + Gd 500 µg l-1

112.12 91.70

Serum + Gd 1000 µg l-1

121.07 91.78

64 March 20, 2014

Titanium in clinical samples

Clinical samples are a complex mix of matrix elements. Of the range of

reaction gases that can be used, NH3 gives good performance for several

key elements in clinical samples, such as Ti (metal-on-metal joints).

Compared NH3 to no gas, He, O2 mode NH3 is a

highly

reactive cell

gas. 48Ti

forms

several

cluster ions

with NH3 cell

gas,

including

TiNH2(NH3)4

TiNH2(NH3)4+

m/z=114

65 March 20, 2014

Q1 set to let in only m/ƶ 46, 47, 48, 49, 50 INDIVIDUALLY

Q2 set to transition masses:

Q1 +84amu [TiNH2(NH3)4]

Q1 +102amu [Ti(NH3)6]

Therefore transitions used:

46 130 46 148

47 131 47 149

48 132 48 150

49 133 49 151

50 134 50 152

MS/MS Neutral Gain Scan – Titanium with

Ammonia Adducts at Two Mass Transitions

66

Q2 set to Q1 +84amu [TiNH2(NH3)4] & +102amu [Ti(NH3)6]

Serum and Urine Check

Based upon results from this HNO3 standard the system was run using basic

preparation for clinical samples (dilution into NH4OH, EDTA, Triton-X, BuOH)

• Standards were prepared in the basic preparation medium

• Standard addition NOT used

• Both diluted serum and urine (10x) run within same batch against same

calibration

• Data compared to no cell gas and other gas modes

Titanium – No Gas, He, O2, NH3

Target 47 -> 131 Ti [ NH3 ] 48 -> 132 Ti [ NH3 ] 49 -> 133 Ti [ NH3 ] 50 -> 134 Ti [ NH3 ]

Sample Name Conc. [ ug/l ] Conc. [ ug/l ] Conc. [ ug/l ] Conc. [ ug/l ]

Urine Blank 4.6 (2.2-7.0) 2.80 2.79 2.92 2.49

Urine Blank 4.6 (2.2-7.0) 3.50 2.93 3.33 2.66

Urine Trace Elements 14.81 15.27 14.42 15.13

Urine Trace Elements 14.99 15.49 15.50 15.05

Serum L1 1.28 (0.86-1.80) 1.21 1.15 1.14 0.80

Serum L1 1.28 (0.86-1.80) 1.27 1.18 1.09 0.89

Serum L2 1.76 1.92 1.61 1.13

Serum L2 1.82 1.64 1.76 1.22

Target 47 Ti [ No Gas ] 47 Ti [ He ] 48 -> 64 Ti [ O2 ]

Sample Name Conc. [ ug/l ] Conc. [ ug/l ] Conc. [ ug/l ]

Urine Blank 4.6 (2.2-7.0) 1989.79 41.44 15.48

Urine Blank 4.6 (2.2-7.0) 2004.91 44.30 15.76

Urine Trace Elements 1789.92 51.41 26.30

Urine Trace Elements 1749.13 52.58 27.08

Serum L1 1.28 (0.86-1.80) 144.18 3.79 7.33

Serum L1 1.28 (0.86-1.80) 128.97 2.95 7.37

Serum L2 100.16 3.95 9.20

Serum L2 95.65 3.02 9.00

“Conventional” Modes (no gas, He, O2)

Ammonia Cell Gas using the +84 transition - TiNH2(NH3)4

NH3 mode gives very consistent recovery for 4 Ti isotopes (47, 48, 49, 50). All results within range of “target”

values, but reference range (all from HR-ICP-MS) is very wide. 8800 ICP-QQQ results on low side of range

67 March 20, 2014

Practical Benefits of Improved Abundance

Sensitivity of ICP-MS/MS

Mn in Fe matrix

Mn in whole blood

B in organics

68

Abundance Sensitivity

69

Abundance

Sensitivity Q1 Q2

Low 5x10-7 5x10-7

High 1x10-7 1x10-7

8800 Abundance Sensitivity in Single-Quad Mode

Trace 55Mn is overlapped by 56Fe in 1000ppm Fe

Abundance Sensitivity (AS) is

the degree of peak “tailing” – the

contribution a peak makes to the

adjacent (-1 and +1amu) masses

7700 specification is 5 x 10-7 on the

low mass side

Note: log intensity scale

70 March 20, 2014

MS/MS Abundance Sensitivity is

the product of Q1 AS x Q2 AS,

so overlaps to adjacent peaks

are virtually eliminated

8800 AS is almost too good to

measure (<10-9)

8800 Abundance Sensitivity in MS/MS Mode 55Mn is free from overlap by 56Fe in 1000ppm Fe

71 March 20, 2014

Other Sample Types – Manganese in Whole Blood

Mn is difficult to measure by ICP-

QMS at natural (sub-ppb) levels in

whole blood, due to “tail” of 56Fe

(and 54Fe) peak across 55Mn

8800 MS/MS ensures 55Mn is

completely separated from 54/56Fe

Overlaid spectra show blank whole

blood (10x dil) & 500ppt Mn spike

72 March 20, 2014

Abundance Sensitivity Test - 1ppm Fe, MS/MS

Mode

73

In a 1ppm Fe standard,

Abundance Sensitivity

is almost too good to

measure, so we

decided to test it

further…

Boron is difficult to measure by ICP-

QMS at trace (low-/sub-ppb) levels

in organic solvents, due to “tail” of 12C peak across major 11B isotope

8800 MS/MS ensures 11B is

completely separated from 12C

Overlaid spectra show blank

kerosene and 5.8ppb B spike (12C is

over-range and skipped)

Practical Applications of Better AS – Boron in

Organics (kerosene)

74 March 20, 2014

Boron in Kerosene BEC & DL Improvement in MS/MS Mode

BEC and DL greatly improved, especially for 11B due to no

contribution from 12C – 11B BEC 140ppt; DL 28ppt

No gas single quad mode No gas MS/MS mode

75 March 20, 2014

ICP-QQQ-MS in the “-omics”

76

ICP-MS as element-specific detector for the life

science applications - Metallomics

•Very sensitive - 1000 times or better than organic mass.

•High ionisation efficiency - good for P, S, Se compounds.

•Compound independent calibration - easy and quick quantification.

•Multi-element capabilities > 30 elements simultaneously

•Wide dynamic range = ppm~ppb~ppt simultaneously

•Isotope ratio measurement - Isotope Dilution, Metal tracer.

•Easily interfaced to multiple separation techniques - On-line detector.

It is recognized that metals play an important roles in proteomics processes as such:

- About 30% of all proteins may bind metal ions.

- There are many proteins known to bind Ca, Mg, Fe, Zn or Ni ions or chromogenic groups like heme or chlorophyll which bind metal ions.

77

Analytical strategy in Metallomics

Separation HPLC, CE

Sample Result

ES-MS/MS MALDI-TOF

Molecular information &

Identification/structure

information

Metal binding information &

Absolute quantification

without specific standards

Extraction/

Purification/

Concentration

Metalloprotein

Sequence

2-D Separation

ICP-MS

79

Absolute Quantitative Proteomics by the ICP-QQQ

The first result from the collaboration with Dr Jorge Ruiz Encinar of Oviedo

University, Spain on the quantitative proteomics by ICP-QQQ has been

obtained and is published in Analytical Chemistry.

80

0

10

20

30

40

50

60

70

80

0

50

100

150

200

250

300

350

400

3,2 3,3 3,4 3,5 3,6

x103

Coun

tsx1

03

Time (min) 34S/32S ratios measured

Injected (Methionine) 0.0489 ± 0.0017

Infusion (sulphate) 0.0487 ± 0.0012

Isotope ratios measured in transient

signal as peak area ratios

32S as 32S16O+

34S as 34S16O+

Mass discrimination factors again

ranging from -4.0 to -4.3%

ICP-QQQ for S isotope ratio analysis

81

Applications Handbook (Publication number 5991-

2802EN)

82

Application Notes

March 20, 2014

83

84 March 20, 2014