孫毓璋國立清華大學

ICP-OES/ICP-MS 分析技術與應用

Trace Analysis

Biogeochemical Cycle of Trace Elements

Atmosphere

Lithosphere

Hydrosphere

Distribution of Trace Elements

Plant Toxic Effects

長芒扁雀麥 (Bromus carinatus)

O'Dell, Silk et.al. 2007

Bioaccumulation of Mercury in Aquatic Food Chain

The fate of trace elements

Environmental Fate Biological Fate

Absorption

Distribution

Transformation

Excretion

Possible Forms of Trace Possible Forms of Trace Elements Elements

Simple ionic species

Differing valency

states

Weak complexes

Adsorbed on colloid

Lipid-soluble

complexes

Organometallic

species

Particulate

Zn(H2O)6+2

As(III), As(V)

Cu-fulvic acid

Cu-Fe(OH)3-humic acid

CH3HgCl

CH3AsO(OH)2

metal adsorbed onto or contained

within clay particles

The nature of spectrometric measurement

Characteristic-photon Characteristic-ion

Cu0 Cu+

Elemental analysis techniques

Analytical ProcedureAnalytical Procedure

Sample Weigh and Decomposition

Sample introductionMeasurement

UrineWhole blood Geological samples

Sea water

Zn,Cd,Pb,Cu

103~10 － 1μg/L

Zn,Cd,Pb,Cu…

103~10 － 1μg/L

Zn,Cd,Pb,Cu…

105~101μg/kg

Zn,Cd,Pb,Cu…

100~10 － 2μg/LTrace

elements

Na, K, Ca, Mg, FeNa, K, Ca, Mg, Fe

0.1~10%0.1~10%

Na, K, Ca, Mg, FeNa, K, Ca, Mg, Fe

0.1 ~10%0.1 ~10%

Fe, Si, Mn, AlFe, Si, Mn, Al

1~30%1~30%

Na, K, Ca, MgNa, K, Ca, Mg

1~10%1~10%

Major composition

Biological samplesEnvironmental samples

Complex MatrixComplex MatrixComplex MatrixComplex Matrix

Environmental Samples•Sea Water•Geological Samples

Environmental Samples•Sea Water•Geological Samples

Industrial Material• High Purity Metals

• Organics

Industrial Material• High Purity Metals

• Organics

Biological Samples•Bontanical Samples•Clinical Samples

Biological Samples•Bontanical Samples•Clinical Samples

Trends and Problems in Trace Element Analysis

Trends

Problems

Ultratrace analysis (in bulk/micro samples)

Micro-distribution

Speciation

In-vivo/In-Situ/ Continuous

Extremely Complex Matrix

- Preconcentration

- Separation

Reliability (Systematic error)

Stability

AAS vs ICPs

6000-8000oC

1200-3000oC

1700-2800oC

Ionization/Atomization

Ionization/Atomization

Ionization/Atomization

ICPs

ETAAS

FAAS

感應耦合電漿

感應耦合電漿反應機制

+

Photon emission(atomic lines)

Photon emission(ionic lines)

excitation source

particle molecule atom ion

liquid sample solid sample

nebulization

introduction of solid sample

aerosol

desolvation

vaporization excitationdissociation

excitationionization

Ionization Potential

Ionization potential ， eV

110

100

90

80

70

60

50

40

30

20

10

0

Se(9.7524)

As(9.7886)

Te(9.0096)

Pb(7.4167)

Sb(8.6084)

Zn(9.3942)

Cd(8.9938)

Cl(12.9676)

S(10.3600)

I(10.4513) Ar

(15.7596)

Cs(3.8939)

Na(5.1391)

Ca(6.1132)

Cr(6.7665)

Mo(7.0924)

Cu(7.7264)

Cd(8.9938)

Ion

izat

ion

eff

icie

ncy

, %

Ionization efficiency of different elements in ICP

Sensitivity

How do they work?

ICP/ optical emissionspectrum of 100 ppm Cerium solution

ICP/ mass spectrumof 10 ppm Cerium

Ce emission

140Ce+

plasma continuum

Spectra obtained by ICP-OES and ICP-MS

Comparison of Detection limits

Table 26-4

Concept-Revolution in ICP-OESConcept-Revolution in ICP-OES

Coventional conceptCoventional concept ICP is an ideal spectrochemical

excitation source that is free from matrix interferences on the analytical line intensity.

Realistic viewRealistic view Different matrix give very complicated

interference on the analytical line intensity

Coventional conceptCoventional concept ICP is an ideal spectrochemical

excitation source that is free from matrix interferences on the analytical line intensity.

Realistic viewRealistic view Different matrix give very complicated

interference on the analytical line intensity

Spectral interferences

54.93494 56.96284

55Fe+:55.93494

111Cd++:55.45209112Cd++:55.95134112Sn++:55.9524140Ar16O+:55.9572940Ca16O+:55.95750113In++:56.45203113Cd++:56.45220

Quadrupole resolution

55.92905 56.96873

ICP-MS ICP-OES―Isobaric―Polyatomic ion ―Matrix-induced molecular species―Doubly charged

―Baseline elevation―Peak overlap

Resolution=10000

Atomization, Excitation and Ionization mechanism in Atomization, Excitation and Ionization mechanism in ICPsICPs

MX

M

M+

M(H2O)m+, X-

Solution

Desolvation

Solid

vaporization

Gas

atomization

Atom

ionization

Ion

(MX)n

excitation

M* hυ

M* hυexcitation

IP

EP

EP

Ionic line=IP+EP

Atomic line=EP

Without MatrixWith Matrix

Non-spectral interferences

待分析元素:鋅

40

50

60

70

80

90

100

110

120

200 400 600

[Matrix element], mg/L

Re

lativ

e s

ign

al,

%

Na

Ca

Fe

Mn

Al

基質元素

Matrix effects on the analytical recoveryMatrix effects on the analytical recovery

CFN-Axially viewed ICP

Energy sum, eV

7.5 9.5 11.5 13.5 15.5 17.5

Al

Mn

Fe

Na

CaRecovery

, arb

itra

ry u

nit

Analytical results of soil leachate

5倍稀釋

0

20

40

60

80

100

120

Be V Cr Co Ni Cu Zn Ga As Se Mo Cd In Sb Cs W Au Hg Tl Pb

Rec

ove

ry,

%

10倍稀釋

0

20

40

60

80

100

120

Be V Cr Co Ni Cu Zn Ga As Se Mo Cd In Sb Cs Ba W Au Hg Tl Pb

Rec

over

y, %

Matrix dilution Internal standard Isotope dilution Standard Addition

Matrix effect vs Ionization Potential

+

Analyte

+matrix

e-

Ar+

Ar2+

Ar+ Ar + 15.75 ev

Ar* Ar + 11.50 or 11.67 ev

Ar2* 2Ar + 10.2 ev

Ar2+ 2Ar +14.03 ev

e- Ar+ Ar2+

Matrix+

Internal StandardizationInternal Standardization

Energy sum, eV

14.03 eV 15.40 eV

Internal standard 1

Internal standard 2

Rec

ove

ry,

%

Analytical results of spiked native soil sample

Analyte Analytical line, nm Energy sum, eV Recovery, % Corrected recovery, %

Sc* 361.384 9.99 95 108

V* 292.402 11.37 102 116

V* 310.230 12.76 100 113

Cr* 205.552 12.8 102 116

Ni# 231.604 14.03 68 99

Cd# 226.502 14.46 76 110

Cd# 214.438 14.77 71 103

Pb# 220.353 14.79 83 110

Zn# 202.548 15.51 76 109

Cu# 224.700 15.96 72 104

Soil Order: Vertisols Spike concentration = 1 mg L -1

* Internal standard = Mo (204.598 nm; energy sum=13.16 eV)# Internal standard = Ir (224.268 nm; energy sum=14.53 eV)

J. Anal. At. Spectrom., 2003, 18, 1163–1170

Analytical results of BCR 141R soil sample

* Internal standard = Mo (204.598 nm; energy sum=13.16 eV)

# Internal standard = Ir (224.268 nm; energy sum=14.53 eV)

Analyte Analytical line, nm Energy sum, eV Recovery, % Corrected recovery, %

Cr* 205.552 12.8 82 95

Ni# 231.604 14.03 69 89

Cd# 214.438 14.77 74 96

Pb# 220.353 14.79 83 108

Zn# 202.548 15.51 68 88

Cu# 224.700 15.51 68 88

J. Anal. At. Spectrom., 2003, 18, 1163–1170

Main Topics in Countercurrent

Analytical Chemistry

New instrumentation

New principle

MiniaturizationAutomization

Bio-chip Micro-fluidic

Chemometrice

On-line analytical system

In-situanalysis

Real-timeanalysis

In-vivomonitoring

Interfacingtechnique

Separation computerization Hyphenation

SensorSingle cellanalysis

Non-destructiveanalysis

Chemicalimagining

Macro-moleculeidentification

Surfaceanalysis

Processanalysis

Environmental &Biological analysis

Arsenic: Evil or Angel?

Napoleon

Arsenic-induced hyperkeratosis

Black foot disease

Poison Acute promyeloid leukemia

Analytical Natural of ICP-MS

Arsenite

Arsenate

Monomethylarsenic

Dimethylarsenic . . .

As+

[M]

Sig

na

lAnalytein 0.5% HNO3

Analytein seawater

GC-ICP-MS: A very sensitive hyphenated system for speciation analysis

http://www.speciation.net/index.html?PHPSESSID=53bca5e65a5918efcd740ec68ab61d97

J. Anal. At. Spectrom., 2008, 23, 861–870

LC-ICP-MS: The most often used hyphenated system for speciation analysis

http://www.speciation.net/index.html?PHPSESSID=53bca5e65a5918efcd740ec68ab61d97

Journal of Chromatography A, 1176 (2007) 143–148

36

Still to be Answered : What cause brain damage in acute ischemic stroke and traumatic stress?

36

In light of the recent observation of stress-related breaks in the blood-brain barrier, the mechanism of metal ion accumulation in the brain should be re-evaluated.

Nature, 1998; 393: 373.

Arch Intern Med 2000;160:3196.

Generalized model of cellular metal ion homeostasis

…in situ studies of cellular metal ion accumulation, trafficking, efflux, and function or toxicity with spatial, temporal and oxidation-state fidelity. …make significant contributions to our understanding of metal ion homeostasis in health and disease.

Nature Chemical Biology 4, 168 - 175 (2008)

Blood flow

Capillary

Extracellular space

J. Anal. At. Spectrom., 2008, 23, 1275–1280

C, Cu and Zn distribution in a cross section of rat brain sample

In-vivo sampling technique – Microdialysis

Cu copper

Al Mn Manganese

Zn zinc

PbLead

FeIron

Menkes' syndrome Wilson's disease

0.9% NaCl

40

Challenges: Trace Amount and Small Volume

Low analyte concentration

10-1 g/L < [Mn+] < 102 g/L

Limited sample volume

Sampling rate=0.5-1 L/min

Sample volume=10 L/20 min

Sensitivity

1 g/L×10 L= 10 ×10-12 g

Selectivity

Normal saline=0.9% NaCl

Target anionic and cationic species

Robustness

12 h continuous monitoring

Blank control

0.9% NaCl

85807570656055504540353025201510510 0

10 1

10 2

10 3

10 4

10 5

10 6

10 7

10 8

10 9

10 10

Mass

ion

sign

al /

cps

O+ Ar+ArH+

ArO+Ar2

+

40

End-determination(ICP-MS)

In vivo monitoring

Microdialysis sampling

On-line desalting(Electrodialyzer)

Microdialysis-On-line desalter-ICP-MS Hyphenated System

Purewater

Anal. Chem. 2007, 79, 8900-8910

42

Simultaneous determination of mercury and selenium in urine by on-line Desalter-ICP-MS system

0

20

40

60

80

100

120

0 0.1 1 2.5 5 10 20

EDTA conc.(mg/L)

reco

very

(%)

Hg

Na+

(EDTA-Hg)-

SeO42-

Hg2+

+EDTA-

Na+

SeO42-

Hg2+

-

+

0

20

40

60

80

100

120

0 0.1 1 2.5 5 10

EDTA conc. (mg/L)

reco

very

(%)

Se 4+Element Certified Found Recovery

Hg (a) 48 ppb 42.4 ± 3.8 ppb 88.3 %

Se (b) 59±5 ppb 59.69±2.4 ppb 101.2 %

(a) SeronormTM Trace Elements Urine(b) NIST CRM No. 18 Human Urine

Removal efficiency of Na+ ion and Recovery of analyte ions

sample solution = 0.9 % NaCl, 650mM NH4Cl pH= 8

元素 / 質量數 回收率 (%)

Zn 66 99.5

Cd 114 99.8

Pb 208 102.5

Cu 65 104.7

Mn 55 93.5

Co59 103.7

Ni 60 105.820

1613

107

4

1

0.5

0.10

20

40

60

80

100

120

Na(%

)去

除效

率

44

Antagonistic effect between Cadmium and Selenium?

44

0

5

10

15

20

25

30

35

40

0 50 100 150 200 250 300 350

0

500

1000

1500

2000

2500

3000

Se

Cd

Time course, min

[Se], u

g/L

[Cd], u

g/L

Cd injection

Injection dose ： 0.8 mg Cd2+/Kg body weight

To Desalter-ICP-MSCd solution

46

How about the permeability of blood-brain barrier ?

46

矽統科技研發部資深經理林廷鴻 ，原本脾氣又壞又暴躁，但去年底出車禍腦部重創，昏迷指數只剩 3 ，竟奇蹟似的醒來，最難能可貴的是，他從暴躁嚴肅的科技人性格，變成把愛掛在嘴邊的好男人，就連太太都說，這真的是「天上掉下來的禮物」。

車禍「洗腦」 大男人變好男人

2007.06.19

47

Causation of the adverse Effects of Cisplatin?

Cisplatin- Used to treat testis cancer

Nausea/vomiting

Kidney damage

Decreased hearing

Decreased sensation

47

48

The Time Course of Mn and Pt in the ECF of the Rat Brain after Intraperitoneal Administration

100 mg MnCl2/kg body weight

2 mg cisplatin/kg body weight

0.9% NaCl

49

Calcium-induced neuro-apoptosis

http://www.benbest.com/cryonics/ischemia.html

Excessive glutamate release resulting in excessive

Ca+2 entry into cells is the excitotoxicity which

initiates the brain ischemic damage seen in stroke

and cardiac arrest.

http://www.fda.gov/nctr/science/journals/text/vol5iss1/rrp0305.htm

http://www.cirp.org/library/pain/anand4/

50

Characterization of Ion Release and Uptake Events of Neural Cell of Living Rat

0

2

4

6

8

10

12

14

16

-300 -200 -100 0 100 200 300 400 500 600 700 800

T ime course, min

[Mg]

, m

g/L

I nsert MD KCl Perfusion Ringer Perfusion

Mg

0

5

10

15

20

25

30

35

-300 -200 -100 0 100 200 300 400 500 600 700 800

T ime course, min

[Ca]

, m

g /

L

Ca

Insert MD KCl Perfusion Ringer Perfusuion

Ca2+

Biol Res 37: 665-674, 2004 /

V V V

V V V

51

00.5

11.5

22.5

33.5

44.5

5

-300 -200 -100 0 100 200 300 400 500 600 700 800

Time course, min

[Mn],

ug/L

Mn

Insert MD KCl Perfusion Ringer Perfusion

Characterization of Ion Release and Uptake Events of Neural Cell of Living Rat

0

4

8

12

16

20

-300 -200 -100 0 100 200 300 400 500 600 700 800

T ime course, min

[Zn

], u

g/L

Zn

I nsert MD KCl Perfusion RingerPerfusion

0

20

40

60

80

100

-300 -200 -100 0 100 200 300 400 500 600 700 800

Time course, min

[Cu],

ug/L

Cu

I nsert MD KCl Perfusion Ringer Perfusion

V V V

V V V

V V V

Thank you for your attention !