ary Flow Focusing Nebulizer (CFFN) for

roscopic Analysis:

cation to Biological Samples

Important Role of Elements

Fe

living
organism

Cr

Zn

tissue cell
protein DNA

essential major elements for human

essential trace elements for human
essential trace elements for mammals

Important Role of Elements

Fe

Zn

Cr

living
organism

tissue cell
protein DNA
sample
kg-g, L mg, mL-L ng, L
size
<ng, nL
major element Fe : 50g/g (ppm) 50 fg
0.9 fmol
trace element Cr : 30 ng/g (ppb)

30 ag
0.6 amol

ag
zmol
zg
ymol

%, g, ppm mol
unit
number

Low sample consumption High sensitivity

Important Role of Elements

Lobinski et al,
Pure Appl. Chem.,
82 (2010) 493504

Simplified model of biological system and related -omics sciences. The outer area surrounded with the
continuous line is showing, e.g., an organ or whole body, and the inner area surrounded with the dotted line
is showing a biological cell. Biological fluid, e.g., blood, is circulating in the intermediate area. The Mg2+ and
Ca2+ ions are given as examples because of their large affinities with DNA and proteins, respectively, in

Analytical Methods for Biological Samples

Fluorescence

X-ray, laser,

(XRF PIXE
etc.)

ablation by laser
Optical emission
radiation
limit of detection
ionization by plasma
ppm ppb
Element
analysis
limit of detection
ppb ppt

Mass
spectrometry

Speciation

hyphenation
with
separation
techniques

(Laser Ablation)

sample solid
distribution, mapping

nebulization
ionization by plasma
Aqueous sample

(Solutionbased)
High Performance Liquid
Inductively
Coupled Plasma Mass Spectrometry ICPChromatography HPLC
MS

A.M. Oros-Peusquensa, A. Matuschb et al., Int. J. Mass Spectrom., 307 (2011) 245-252

Sample Introduction in Typical ICP-MS

sample introduction

plasma gas

cooling gas

ionization
plasma

plasma torch

mass spectrometry

carrier gas

drain

nebulizer

spray chamber

carrier gas

to plasma
<10 mm

>10 mm

95

sample
0.1 1 mL/min

to drain

conventional system
High sample aspiration rate & Low sample introduction efficiency
much sample loss unsuitable for small sample
volume
dilution undesirable for low concentration of

commercially available high efficiency nebulizer

sensitivity
micronebulizer

ultrasonic nebulizer

nebulizer tip clogging

Sample

high memory effect

long washing time

sample consumption
micro plasma

low energy
low efficiency
(change of plasma form)

Direct-Injection
Nebulizer

matrix effect
complicated handling
(gas flow rate, pressure)

Development of High Efficiency Nebulizer?

5
sample

small amount of
biological sample

95

conventional system

Highly Efficient Nebulizer

Low sample uptake, Total sample consumption
Easy handling, Salt tolerance
Facile combination with flow-based techniques
Generate fine and monodisperse aerosol droplets

Theoretical Aspects of Nebulization

nebulizer tip configuration

Hagen-Poiseuille

low
pressure

dt 8
dV

We

1.5

20 50 100
CFF

1000

TFF

r tube radius, fluid

viscosity
p pressure drop, l tube
length
H

Ganan-Calvo

Stable flow
0.5 H 1. 5
D

H: tube-orifice distance
D: tube orifice diameter

Weber (We )

g ug2 dj
2

0.5
0.1

FB
CFF: capillary flow focusing
TFF: turbulent flow focusing
FB: flow blurring

: density, u; velocity,
d: jet diameter, : surface tension

J. Rosell-Llompart, A. M. Gan-Calvo, Phys. Rev. E, 77 (2008) 036321

Schematic of the processes taking place at the exit of

a concentric pneumatic nebulizer (CPN)

CPN

dV r p

dt
8 l
4

(dV/dt): volume flow rate of a homogeneous

fluid passing through a tube
(p): pressure drop
(r): tube radius (r)
(l) the tube length
() fluid viscosity

Development of New Micronebulizer

0.5 H 1. 5
D
1 We
20

Capillary Flow Focusing

Nebulizer (CFFN, home-made)
nebulizer body

PEEK
connector

fitting

capillary tube

tubing
sleeve

commercially available nebulizer

Conikal (1mL/min)
H 2.32
D ,

capillary tube

We 80.7

MicroMist (100L/min)
H
D 1.75 ,

nebulizer body

We 25.5

center nozzle
Capillary: 75 m i.d./ 150 m o.d.
Orifice : 50 m i.d./ 80 m o.d.

Capillary Flow Focusing Nebulizer (CFFN)

We

Capillary Flow Focusing Nebilizer

CFFN (
10L/min)
H

We 8.08
D 1.33 ,
capillary flow focusing

1.5

H
D

20 50 100
CFF

1000

TFF

0.5
0.1

FB
CFF: capillary flow focusing
TFF: turbulent flow focusing
FB: flow blurring

Droplet Size Distribution of Aerosols

commercially available nebulizer
(MicroMist)

30

30

20

20

Distribution [%]

Distribution [%]

Capillary Flow Focusing Nebulizer

10
0
1

10
Droplet Diameter [m]

100

10
0
1

10
Droplet Diameter [m]

100

Free Aspiration Rate

Operating conditions of ICP-MS
cooling gas : 13.00 L/min
rf power : 1.28 kW
carrier gas : 0.96 L/min
plasma gas : 0.93 L/min
free aspiration rate of CFFN
free aspiration rate of commercially available nebulizer

0.1 M HNO3

20

100
< 10L/min

0
75

100

150

200

Inner diameter of capillary [m]

250

300

Free aspiration rate

200

[L/min]

80

1000

Decrease in the pneumatic

free aspiration rate

Sample Introduction Efficiency (SIE)

aspirated [L]-drained [L]
[ ]
aspirated

100 [L]

SIE

free aspiration rate by CFFN

introduction efficiency by CFFN
free aspiration rate by commercially available nebulizer
introduction efficiency by commercially available nebulizer

0.1 M HNO3

Introduction efficiency
[]

95

60

200

40
20
0

100
< 10L/min

0
75

100

150

200

Inner diameter of capillary [m]

250

300

Free aspiration rate

80

1000

Increase in the sample

introduction efficiency

[L/min]

100

Thickness of Capillary Tube (Outer

nebulizer body

diameter)

nebulizer body

sample volume : 50L

flow rate
: 500L/min
inner nozzle
concentration : 1 ppm 25.5 m

60
Fe
Ni
inner
103
Rh nozzle185Re

P
66
Zn

56

31

capillary tube

63

Cu

138 m

75 m i.d.
polyimide coating

i.d. 75
m
o.d. 150
m

Signal intensity [cps]

Signal intensity [cps]

108
107
106
105
104
103
102
101
100

i.d. 75
m
o.d. 375
m
Growing
&

50

100

Time [min]

150

RSD 2.11 4.19

200

108
107
106
105
104
103
102
101
100

Renebulization

50

100

Time [min]

150

RSD 6.08 7.18

200

Effect of Spray Chamber Temperature

temperaturecontrollable
spray chamber
(IsoMist)

desolvation vaporization
atomization ionization
M
M

60

2104

1.5

0.5
0

3104

P+
32 +
S
56
Fe+
63
Cu+
66
Zn+
111
Cd+
31

0 10 20 30 40 50 60 70
Chamber temperature [ C]
0

Signal intensity [cps]

Relative intensity

2.0

1.0

1104

twice

0
0

100
Time [sec]

200

3104

25

2104
1104
0
0

100
Time [sec]

200

Flow Injection System-CFFN

carrier
solution
0.1M HNO3

20 L
CFFN

metal-free pump
injector

spray chamber
(IsoMist)
60

Signal intensity of 111Cd

[cps]

10 L/min

ICP-MS
(Element 2)

sample : 10 g/L Cd, 20 L

carrier solution : 0.1M HNO3, 10 L/ min
45000
40000
35000
30000
25000
20000
15000
10000
5000
0
0

continuous flow
measurement
time

20 L loop
injection

signal
stability
(RSD)
1.2%
1.5%

washing time
50

100

Time [sec]

150

200

Flow Injection System-CFFN

Signal intensity of 111Cd

[cps]

Dispersion
coefficient

(D)

signal intensity of the continuous

(C0)
sample introduction
signal intensity by the
loop injection

(Cmax)

1.02

sample : 10 g/L Cd, 20 L (5 injections)

carrier solution : 0.1M HNO3, 10 L/min
45000
40000
35000
30000
25000
20000
15000
10000
5000
0

repeatability
RSD 1.5%

200

400

Time [sec]

600

800

1000

Repeatability
without

sample : 20L flow rate : 10L/min

concentration : 1 ppm
10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0
10 0

Signal Intensity / cps

10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0
10 0
10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0

10 0
10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0
10 0
10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0
10
0
10 87
10 6
10 5
10 4
10 3
10 2
10 1
10 0
10 0

with

internal standard

P : 2.6 1.5

31

20

40

60

80

100

Ni : 2.7 1.0

60

20

40

60

80

100

Cu : 2.8 0.7

63

20

40

60

80

100

66
20

40

60

80

100

103
20

40

60

80

Rh : 2.9

100

Re : 2.7

185

20

40

60

Time / min

80

Zn : 2.6 1.2

100

Comparison of the Detection Limits

Absolute detection limit
[pg]

103
102
Proposed system (CFFN)

101
100
10-1
10-2
10-3

Conikal nebulizer

Cr Mn Fe Co Ni Cu Zn Cd Pb

Proposed system (CFFN),

Sample: 20 L

Micronebulizer/TISIS,
Sample: 200 L

Micronebulizer/disolvator system,
Sample: 1000 L

d-DIHEN,
Sample: 100 L

Comparison of the Detection Limits

Element

Concentration LOD
[ng/mL]
Proposed

Na
P
S
Fe
Cu
Zn
Cd

Sample
volume

-Flow Injection/FI -CFFN

Proposed system

Conventional

0.2
0.012
0.3
0.02
0.007
0.02
0.0006

0.5
0.014
0.2
0.02
0.006
0.02
0.0007

20 L

2 mL

commercially available
Conikal nebulizer/free aspiration
Conventional system

Comparison of the Detection Limits

Element

Concentration LOD
[ng/mL]
Proposed/
CFFN

Na
P
S
Fe
Cu
Zn
Cd

sample
volume

Conventional

0.2
0.012
0.3
0.02
0.007
0.02
0.0006

0.5
0.014
0.2
0.02
0.006
0.02
0.0007

20 L

2 mL

Absolute LOD
[pg]
Proposed/
CFFN
4
0.2
6
0.4
0.14
0.4
0.012

SEF :
Sensitivity
enhancement
Conventional
factor
1000

28
400
40
12
40
1.4

250

117
67
100
86
100
117

15

Validation of -FI-CFFN-ICP-MS
sample : NIST SRM 1577b (Bovine liver), volume : 20 L,
carrier solution : 0.1 M HNO3, 10 L/ min
Element

Na
P
S
K
Ca
Mn
Fe
Co
Cu
Zn
Mo
Cd

Measured value
[g/g]
2620
11400
7820
9700
108
10.9
192
0.243
172
137
3.84
0.553

30
100
70
120

0.3

0.011

0.11

0.002

RSD
Certified value
[%]
[g/g]
1.0
0.9
0.8
1.2
2.1
2.9
2.5
4.6
2.9
2.4
2.9
0.3

Ratio*

60
2420
1.08
300
11000
1.04
60
7850
1.00

9940
20
0.98

116
4
0.93
10.5
1.7
1.04
15
184
1.04
0.25 (Information value) 0.97

160
8
1.07
16
127
1.08
3.5
0.3
1.10
0.50
0.03
1.11
* Measured value/Certified value

drastic downsizing of sample consumption

without sacrificing accuracy and precision

16

Application to Multielement Analysis

Amount of element [ / cell]

sample : Escherichia coli (MG1655) digested

carrier solution : 0.1M HNO3, 10 L/ min
108

proposed system
conventional system

107
106

Escherichia coli
Gram-negative
Anaerobic rod
Non-sporeforming
0.5(2.0-5.0)m

105
104
103
102
101
100

Na P Fe S K MgCa Zn Ni Ba Pb Cr MnCu Y

nebulizer

introduction rate

sample volume

cells

conventional Conika
l

1000 L/min

2000 L

105 cells

10 L/min

20 L

103 cells

CFFN

17

Estimation of the Distribution of

Stem Cells in Mice Organs

Quantum dots (QDs) are inorganic probes that consist of

CdSe/ZnS-core/shell
semiconductor
nanocrystals
(tiny
nanoparticles) and have recently been explored as
fluorescent probes for stem cell labeling.

Adipose Tissue-derived Stem Cells (ASCs)

Less invasive harvest

procedure
Easier to get in larger
volume
ASCs
Absence of rejection

Curative option
for liver
diysfunction

Distribution in
organs ?

QDs labeling

fluorescence
imaging

core CdSeTe
shell ZnS
polymer coating
Quantum dots
Qdot655

intravenous
injection

H. Yukawa et al., Cell Transplant., 18 (2009) 611-618

H. Yukawa et al., Biomaterials, 31 (2010) 4094-4103

QDs-labelled ASCs

Intracellular distribution and fluorescence intensity of QDs-ASCs.

(Data obtained by Confocal Laser Scanning Microscopy)

Adipose Tissue-derived Stem Cells (ASCs)

QDs labeling

Less invasive harvest

procedure
Easier to get in larger
volume
ASCs
Absence of rejection

Curative option
for liver
diysfunction

fluorescence
imaging

core CdSeTe
shell ZnS
polymer coating

normal vessel
volume 100 mL
vessel insert
5 mL

micro vessel
volume

intravenous
injection

Quantum dots
Qdot655

acid volume

volume

Distribution in
organs ?

1 mL

10 mL
1 mL

100L

sample

instrument

heating 40 min

cooling 120 min

100 mg
10 mg

1 mg

required time

ETHOS E
1000 W

domestic microwave
oven 500 W

heating 30 min

cooling 60 min
heating 5 min

cooling 5 min

10

Cd

Te
QD core

ASCs

QDs-ASCs

Se

ASCs
QDs-ASCs

QDs-ASCs
ASCs

n.d.

QDs-ASCs

n.d.

ASCs

101

ASCs
QDs-ASCs

102

ASCs

103

QDs-ASCs

104

[ f mol/cell ]

Amounts of elements in ASCs

Components of ASCs and QDs-ASCs

Zn

QD shell
Quantum dots (QDs)

Microwave-assisted digestion procedure

for limited samples
sample : NIST SRM 1577b (Bovine liver), volume : 20 L,
carrier solution : 0.1M HNO3, 10 L/min
Limit of detection
[sample: 1
mg]
[sample: 10 mg]

Measured/Certified value

[sample:
[sample: 1
10mg]
mg]

Concentration [g/g]

106

1.2
1.0

103

0.8
100
10-3
10-6
0
Na Mg P S K Ca Mn Fe Co Cu Zn Se Sr Mo Cd Te

Measured / Certified value

Measured value
[sample: 1
mg]
[sample: 10 mg]

Sample Preparation
8 nM QDs

normal mice

liver-injury mice
collected from female
C57BL/6 mice and cultured
5105 cells (150L)

macropinocytosis

QDs-ASCs-treated liver-injury mice

1. major organs were

harvested

2. washed with high

purity water

5. digested
(microwave-assisted acid digestion)

3. Freeze dried

4. crushed and
homogenized

ICP-MS measurement

Multielement Analysis of Mice Organs

106

106

Heart

106

104

Lung104

102

102

102

100

100

100

10-2

10-2

10-2

10-4

10-4
Na P K Fe Zn Co Sr Cd
Mg S Ca Cu Mn Se Mo Te

10-4
Na P K Fe Zn Co Sr Cd
Mg S Ca Cu Mn Se Mo Te

106

106

concentration [ng /g]

104

104

Spleen104

102

102

100

100

10-2

10-2

10-4

10-4
Na P K Fe Zn Co Sr Cd
Mg S Ca Cu Mn Se Mo Te

Liver

Na P K Fe Zn Co Sr Cd
Mg S Ca Cu Mn Se Mo Te

Kidney

normal
liver-injury
QDs-ASCs-treated
liver-injury

Na P K Fe Zn Co Sr Cd
Mg S Ca Cu Mn Se Mo Te

16 elements including essential elements and major

components
of QDs were successfully determined
the concentration of Cd and Te varied among organs

Distribution of ASCs in Mice Organs

Organs

Cd
Te
Molar Ratio of
Cd/Te
[pmol] [pmol]
Heart
39.3
1.67 27
Lung
1270
53.2 26
Liver
2450
106 27
Spleen
32.3
1.3425
27
Kidney
16.7
0.759
QDs-ASCs
6660
281 27

Others Distribution Liver

42.8% of ASCs 36.8%
Lung
19.1%

the incremental molar ratio of Cd/Te Kidney

0.25%
was in fair agreement with the molar
Spleen Heart
composition ratio in QDs-ASCs
0.49% 0.59%
Cd and/or Te can be used
as an accurate tracer for ASCs

Quantification of DNA by
HPLC/ICP-MS Hyphenated
System

cell

Construction of HPLC/ICP-MS
Hyphenated System
HPLC/UV : separation
of biomolecules

ICP-MS : detection/quantification
of multielement
good separation ability as well as small sample consumption
highly sensitive and accurate detection
hyphenation of HPLC with ICP-MS
is an effective approach for biomolecules quantification
samples for evaluation : DNA,
Oligothymidilic acid (dT12-dT18)

Construction of HPLC/ICP-MS
Hyphenated System
keeping good resolution, sensitivity, and plasma stability
matching the flow rate of HPLC with the aspiration rate of ICPMS
inhibiting salt clogging

Capillary Flow Focusing

Nebulizer

Monolithic column

salt clogging

Low flow-resistant
Excellent mass transfer

commercially
available

CFFN

Excellent sample introduction efficiency

Salt-tolerance

Optimization of separation parameters

Eluent: tris-HCl (20 mM) + 1 M salt)
Salt

effectiveness

sensitivity

stability

NaCl

decrease to
1/10

plasma
instability

NH4Cl

pH

dT12
dT13
dT14
dT15

pH 9.0

dT12

pH 8.0

dT18
Sample:
oligodeoxythymidilic acids
(mixture of dT12 dT18)

pH 7.0

dT16
dT17
dT18

pH
7.8

8
7
6

pH 6.0
Mobile phase
(A):20mM Tris-HCl
(B):1M NH4Cl in (A)

NaClaq

9
pH

dT13

Water

10
20
30
Retention time [min]

10

15
20
Retention time [min]

25

HPLC/ICP-MS for quantification of Oligonucleotides

Sample: oligodeoxythymidilic acids mixture of dT12 dT18
Mobile phase(A):20mM Tris-HCl (pH 7.8), (B):1M NH4Cl in 20mM Tris-HCl
Gradient elution: 0% 75% B in 25 min

P-selective chromatogram
by HPLC/ICP-MS
Signal intensity of 31P
[cps]

200000

dT12

dT18

dT18
dT17
dT
dT13 14

dT12

150000
100000

50000
0
0

10
Time / min

15

20

LOD of dT18

250.53 fmol/L

Quantification of Oligoucleotides
dT sample
[mer]
PO , nucleotides
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
34

P mass fraction
[ng/L]
0.0290
0.0685
0.192
0.455
0.547
0.596
0.874
1.19
1.29
1.48
1.49
7.51
7.78
6.10
8.23
5.19
7.88
7.23

0.0021
0.0005
0.005
0.014
0.010
0.008
0.003
0.02
0.02
0.03
0.03
0.28
0.30
0.23
0.31
0.19
0.30
0.27

dT mass fraction
[ng/L]

0.604
1.75
4.23
5.15
5.65
8.33
11.4
12.3
14.3
14.4
72.4
75.1
59.0
79.7
50.3
76.4
70.2

LOD of dT
[fmol/L]

0.004
0.04
0.13
0.10
0.08
0.03
0.2
0.2
0.3
0.3
2.7
2.9
2.2
3.0
1.9
2.9
2.7

4.8
3.2
2.4
1.9
1.6
1.4
1.2
1.1
0.96
0.87
0.80
0.74
0.68
0.64
0.60
0.56
0.53

HPLC/ICP-MS Analysis of DNA Samples

sample : 20 bp DNA ladder mixture of 20 bp 500 bp
mobile phase (A):20mM Tris-HCl (pH 7.8), (B):1M NH4Cl in 20mM Tris-HCl
gradient : 65% - 75% B in 7 min, 75 - 80% B in 11 min, 80% B for 2 min

P-selective chromatogram
by HPLC/ICP-MS

500 bp

1 base pair

500 bp

5
10
Time [min]

15

400 bp

300 bp

120 bp
140 bp
160 bp
180 bp

2000

200 bp

100 bp
40 bp

4000

60 bp
80 bp

6000

20 bp

8000

PO43- + nucleotides

Signal intensity of 31P

[cps]

10000

200 bp
40 bp
20 bp

100 bp

20 LOD (100 bp DNA)

0.048 fmol/L

Separation and sensitive detection of DNA sample

were successfully performed

Quantification of DNA Fragments

DNA sample
[base pairs]
PO43-, nucleotides
20
40
60
80
100
120
140
160
180
200
300
400
500
total
manufactures
value

P mass fraction
[ng/L]
13.7
2.37
1.25
0.920
1.06
2.28
0.992
1.01
1.01
0.720
2.42
1.17
1.01
3.02

0.5
0.06
0.01
0.003
0.002
0.05
0.0003
0.0002
0.0005
0.012
0.06
0.007
0.0003
0.08

DNA mass fraction

[ng/L]

24.8
13.1
9.64
11.1
23.9
10.4
10.5
10.6
7.55
25.4
12.3
10.6
31.6
201
200

LOD of DNA
[fmol/L]

0.6
0.1
0.03
0.02
0.6
0.004
0.002
0.01
0.12
0.6
0.07
0.003
0.9

0.24
0.12
0.080
0.060
0.048
0.040
0.034
0.030
0.027
0.024
0.016
0.012
0.0096

Construction of HPLC/ICP-MS
Hyphenated System
keeping good resolution, sensitivity, and plasma stability
matching the flow rate of HPLC with the aspiration rate of ICPMS
inhibiting salt clogging

Capillary Flow Focusing

Nebulizer

AEX-Monolithic column

salt clogging

Low flow-resistant
Excellent mass transfer

commercially
available

CFFN

Excellent sample introduction efficiency

Salt-tolerance

HPLC/ICP-MS Analysis of DNA Samples

sample : 20 bp DNA ladder mixture of 20 bp 500 bp
mobile phase (A):20mM Tris-HCl (pH 7.8), (B):1M NH4Cl in 20mM Tris-HCl
gradient : 65% - 75% B in 7 min, 75 - 80% B in 11 min, 80% B for 2 min

P-selective chromatogram
by HPLC/ICP-MS

500 bp

1 base pair

500 bp

5
10
Time [min]

15

400 bp

300 bp

120 bp
140 bp
160 bp
180 bp

2000

200 bp

100 bp
40 bp

4000

60 bp
80 bp

6000

20 bp

8000

PO43- + nucleotides

Signal intensity of 31P

[cps]

10000

200 bp
40 bp
20 bp

100 bp

20 LOD (100 bp DNA)

0.048 fmol/L

Separation and sensitive detection of DNA sample

were successfully performed

Quantification of DNA Fragments

DNA sample
[base pairs]
PO43-, nucleotides
20
40
60
80
100
120
140
160
180
200
300
400
500
total
manufactures
value

P mass fraction
[ng/L]
13.7
2.37
1.25
0.920
1.06
2.28
0.992
1.01
1.01
0.720
2.42
1.17
1.01
3.02

0.5
0.06
0.01
0.003
0.002
0.05
0.0003
0.0002
0.0005
0.012
0.06
0.007
0.0003
0.08

DNA mass fraction

[ng/L]

24.8
13.1
9.64
11.1
23.9
10.4
10.5
10.6
7.55
25.4
12.3
10.6
31.6
201
200

LOD of DNA
[fmol/L]

0.6
0.1
0.03
0.02
0.6
0.004
0.002
0.01
0.12
0.6
0.07
0.003
0.9

0.24
0.12
0.080
0.060
0.048
0.040
0.034
0.030
0.027
0.024
0.016
0.012
0.0096

Thank You

Flow Injection System

Signal intensity of 111Cd

[cps]

Dispersion
coefficient

(D)

signal intensity of the continuous

(C0)
sample introduction
signal intensity by the
loop injection

(Cmax)

1.02

sample : 10 g/L Cd, 20 L (5 injections)

carrier solution : 0.1M HNO3, 10 L/min
45000
40000
35000
30000
25000
20000
15000
10000
5000
0

repeatability
RSD 1.5%

200

400

Time [sec]

600

800

1000

Effect of Spray Chamber Temperature

temperaturecontrollable
spray chamber
(IsoMist)

desolvation vaporization
atomization ionization
M
M

60

2104

1.5

0.5
0

3104

P+
32 +
S
56
Fe+
63
Cu+
66
Zn+
111
Cd+
31

0 10 20 30 40 50 60 70
Chamber temperature [ C]
0

Signal intensity [cps]

Relative intensity

2.0

1.0

1104

twice

0
0

100
Time [sec]

200

3104

25

2104
1104
0
0

100
Time [sec]

200

Free Aspiration Rate

Operating conditions of ICP-MS
cooling gas : 13.00 L/min
rf power : 1.28 kW
carrier gas : 0.96 L/min
plasma gas : 0.93 L/min
free aspiration rate of CFFN
free aspiration rate of commercially available nebulizer

0.1 M HNO3

20

100
< 10L/min

0
75

100

150

200

Inner diameter of capillary [m]

250

300

Free aspiration rate

200

[L/min]

80

1000

Decrease in the pneumatic

free aspiration rate

Microwave-assisted digestion procedure

for limited samples
sample : NIST SRM 1577b (Bovine liver), volume : 20 L,
carrier solution : 0.1M HNO3, 10 L/min
Limit of detection
[sample: 1
mg]
[sample: 10 mg]

Measured/Certified value

[sample:
[sample: 1
10mg]
mg]

Concentration [g/g]

106

1.2
1.0

103

0.8
100
10-3
10-6
0
Na Mg P S K Ca Mn Fe Co Cu Zn Se Sr Mo Cd Te

Measured / Certified value

Measured value
[sample: 1
mg]
[sample: 10 mg]

Adipose Tissue-derived Stem Cells (ASCs)

QDs labeling

Less invasive harvest

procedure
Easier to get in larger
volume
ASCs
Absence of rejection

Curative option
for liver
diysfunction

fluorescence
imaging

core CdSeTe
shell ZnS
polymer coating

normal vessel
volume 100 mL
vessel insert
5 mL

micro vessel
volume

intravenous
injection

Quantum dots
Qdot655

acid volume

volume

Distribution in
organs ?

1 mL

10 mL
1 mL

100L

sample

instrument

heating 40 min

cooling 120 min

100 mg
10 mg

1 mg

required time

ETHOS E
1000 W

domestic microwave
oven 500 W

heating 30 min

cooling 60 min
heating 5 min

cooling 5 min

Separation of Oligonucleotide
dT30
Linear gradient elution

100
80

10

20

30

40

50

60

Retention Time (min)

Flow rate: 0.05mL/min(0.5 MPa)

A 20 mM Tris-HCl
(pH 8)

60

40

40

20

20

Sample:
TTTTTTTTTT
TTTTTTTTTTT
TTTTTTTTTTTT

0.2 L

10

Flow rate: 0.25 mL/min(2.6 MPa)

O
NH
+

0.5 MPa

OH

0.012 mL min-1

B A 1M
NaCl

14

Retention Time (min)

0.012 mL min-1

0.5 MPa

12

Size: 1.0 mm i.d x 10 cm260

longnm
T : 270C

B.Conc

80

dT30

B.Conc

60

dT10

100

Shallow gradient elution

dT10