la scienza delle misure chimiche e qualità della vita mario gallorini i.n.r.i.m unità di...

La scienza delle misure chimiche e qualità della vita

Mario Gallorini

I.N.R.I.M Unità di Radiochimica e Spettroscopiac/o Università degli Studi di Pavia

Torino - 25 Gennaio 2007

METROLOGY ?

ENVIRONMENTAL

POLLUTION

Food Chain

ANALITICAL

CLINICAL CHEMISTRY

NUTRITION

DIAGNOSIS

DRUGS

HUMAN HEALTH

TERAPY

COSMETICS

INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia

TRACE METALS EXPOSURE AND HEALTH EFFECTS

LEVELS, FORMS AND

TOXICOLOGICAL SIGNIFICANCE OF TRACE METALS IN HUMANS

MICRODETERMINATION OF TMs IN BIOLOGICAL

SPECIMENS

METABOLISM

(current levels)

TMs IN DIFFERENTLY EXPOSED HUMANS

PATHWAYS

ANALYTICAL QUALITY

ASSURANCE STUDIES BIOCHEMICAL MECHANISMS

TRHESHOLDS LIMITS

RULES ENFORCEMENT


NUCLEAR AND RADIOANALYTICAL TECHNIQUES IN

TRACE METALS ENVIRONMENTAL POLLUTION AND HUMAN IMPACT

SOURCES, DISTRIBUTION, TRANSPORT, EXPOSURE (ENVIRON. PROFESSIONAL…) EPIDEMIOLOGICAL, CLINICAL CASES, CURRENT LEVELS (BASAL CONCENTRATIONS) MAXIMUM PERMISSIBLE DOSES

QUALITATIVE AND QUANTITATIVE DOSE RESPONSE RELATIONSHIP (CHEMICAL FORMS, SPECIES)

KINETIC OF DISTRIBUTION, METABOLISM, METALLOBIOCHEMISTRY ( INTERACT. WITH PROTEINS, ENZYMES…) BIOCHEMICAL EFFECTS AND TOXICITY

MICROANALYSIS

INAA-RNAA

RADIOANALYTICAL TECHNIQUES, RADIOTRACERS, RADIOLABELLED CHEMICAL FORMS, IN VIVO - IN VITRO EXPERIMENTS


Analyses on more than 350 subjects Routine analytical techniques: ET-AAS (Electrothermal Atomic Absorption Spectroscopy)

ICP-AES (Inductively Coupled Plasma Emission Spectroscopy ICP-MS (Inductively Coupled Plasma Mass Spectrometry)

Reference analytical techniques: INAA (Instrumental Neutron activation analysis) RNAA (Radiochemical Neutron Activation Analysis)

“BASAL” CONCENTRATION VALUES OF SOME TRACE ELEMENTS DETERMINED IN WHOLE HUMAN BLOOD g/L

Element Mean Reference Values (range)

Element Mean Reference Values (range)

As 7.90 1.75 0.4 –11.9 Au 0.045 0.0007

0.006 – 0.049

Bi 0.49 0.02 0.12 – 0.8 Eu 0.21 0.08 0.005 – 0.5 Cd 0.60 0.30 0.1 – 1.7 Yb 0.15 0.08 0.05 – 0.3 Co 0.39 0.13 0.01 – 0.9 Th 0.21 0.1 0.03 – 0.6 Cr 0.23 0.01 0.01 – 0.45 Sm 0.26 0.14 0.03 – 0.45 Hg 5.30 0.95 1.7 – 9.9 Sc 0.06 0.015 0.002 – 0.12 Mn 8.80 0.20 7.1 – 10.5 Ir 0.007 0.004 0.0002 – 0.02 Ni 2.30 0.16 1.3 – 3.3 U < 0.1 - Pb 157 9.9 39 - 275 Pt <0.1 - Sb 2.16 0.45 0.03 – 3.5 Rh <0.1 - Se 107 6.4 76 - 140 Pd <0.1 - Tl 0.39 0.05 0.15 – 0.63 V 0.35 0.11 0.09 – 0.75

W 0.39 0.15 0.05 – 0.70 Cu 1225 64 807 - 1643 Zn 6340 210 4076 - 7594 Rb 2805 408 900 - 4145 Cu 1225 64 807 - 1643


EUROTERVIT - JRC- Ispra 1990-2000

TRACE ELEMENTS DETERMINATION IN BIOLOGICAL SPECIMENS AT NANOGRAM OR SUB-NANOGRAM

LEVELS

REQUIREMENTS

HIGH SENSITIVITY

- microsamples - ppb or fractions

PRECISION , ACCURACY

- “normal” levels - narrow window (essential./toxicity)

CHEMICAL METROLOGY

- protocol: methods, reproducibility, primary

standards, calibration, uncertainties...

- quality assurance: certified reference materials

DATA AND RELATIVE SD


TRACE ELEMENTS ANALYSIS : PRE-MEASUREMENTS KEY POINTS

PRE- SAMPLING

- PURPOSE

- TYPE OF SAMPLE

- WHERE AND WHEN

(representativeness)

SAMPLING (contamination / losses)

- TECHNIQUES AND METHODS

- EQUIPMENTS AND MATERIALS

(contamination / losses) STORAGE

- PROCEDURES (freeze-drying, liophyl., …)

- VARIATION (moisture, weight, degradation, homogeneity, chemical forms.. )

DISSOLUTION

- METHODS, EQUIPMENTS

- REAGENTS, CHEMICALS BLANKS

(contamination / losses)

PRETREATEMENTS

- PRECONCENTRATION , SEPARATION…. - PRE- CHEMISTRY

BLANKS (contamination / losses)

FINAL MEASUREMENTS


TRACE METALS DETERMINATION IN BIOLOGICAL SPECIMENS

SAMPLING

Procedure used to evaluate possible TM contamination

during samples collection

A

Characterization and analysis of the materials

which come in contact with the samples

Identification of the potential contaminants

B

Neutron activation of the materials (bistoury,

needles, forceps

Production of the corresponding contaminants

radionuclides

C

Contact of the irrad iate materials with the samples

(radiorelease method)

Identification of the

contaminats

D

NAA of “uncontamined” sample in comparison with the same sample treated with the sampling

materials

Quantification of the degree of contamination

Trace metals released from surgical tools in collecting rabbit lung tissues.

Radio-release procedure.

Element bistoury (ng)

forcep (ng)

human lung w.t.

(ref. Value ng/g)

Ag 1.3 120 2.1

Ce 25 80 87

Co 3.7 25 80

Cr 50 3200 312

Cs 0.7 1.3 17.2

Fe 1700 30000 2640000

Hg < 10 17 90

Ir 0.02 0.02 0.2

Ni 135 450 600

Rb 37 140 4775

Sb 22 35 230

Ta 2 14 13

Zn <30 120 12130


TRACE METALS DETERMINATION IN BIOLOGICAL SPECIMENS

BLOOD/SERUM SAMPLING

Vanadium contamination from commercial syringes

ng of V

needle 0.03-0.09

“ICOGAMMA” insuline cylinder 0.13

stopper 0.08

needle 0.05

“Pronto Siringa” cylinder 0.62

stopper 0.10

V reference value in whole human blood : 0.09 – 0.75 g/L

Syringe and needle irradiated neutron activated (6·10·12 n·cm-2·s-1) and leached with 20 mL of human serum

Element

stainless needle syringe (stopper +cylinder)

Human Serum Reference Values

g/L

Maximum amount released g/L

Ag <0.05 <0.05 0.18 As 0.8 3.1 7.9 Au <0.01 <0.01 0.01 Ba 0.5 12.7 1.2 Cd <0.1 2.4 0.2 Ce <1 <1 3.1 Co 5 0.7 0.21 Cr 12 0.6 0.17 Cs <0.1 <0.1 1.5 Eu <0.1 <0.1 0.21 Hf <0.1 <0.1 0.21 Hg <0.1 <0.1 2.1 Ir <0.001 <0.001 0.007 Mn 1.1 <0.1 0.6 Mo 26 12 - Rb <10 <10 230 Sb <0.1 4 0.5 Se <2 <2 81 Ta <0.1 <0.1 0.23 W 1.2 <0.1 0.39 V 0.7 0.1 0.65 Zn 6 18 922


TRACE METALS DETERMINATION IN BIOLOGICAL SPECIMENS human tissues SAMPLING

Note: relative SD from 10 to 15%

Trace Metals release into human blood of uncoated and TiN coated AISI 316 stainless

steel surgical blade

metal release (ng)

“Antirelease” Coefficient ARC

TiN coated vs. uncoated surgical blade

Element

Uncovered

AISI 316 blade

TiN-coated AISI 316 blade

Pure TiN

(50 mg)

Element

ARC

Al 200 < 50 < 20 Al 4

As 0.3 < 0.3 < 0.2 As 1

Au 0.5 0.3 < 0.01 Au 1.6

Cd 10 < 3 < 1.5 Cd 3.3

Co 12 3 < 3.5 Co 4

Cr 640 100 35 Cr 6.4

Cs 4.6 1.6 < 3.5 Cs 2.8

Fe 2000 800 760 Fe 2.5

Hg 0.5 <0.2 < 0.3 Hg 2.5

La 0.65 0.15 < 2.5 La 4.3

Mn 200 0.8 < 1 Mn 250

Mo 0.8 < 0.5 < 50 Ni 10.7

Ni 3000 280 360 Sb 3.4

Sb 5.1 < 1.5 4.0 Sc 1.8

Sc 0.09 0.05 < 0.02 Ta 3.4

Se 0.5 0.5 < 0.5 Ti 3

Ta 1.7 0.5 < 0.2 V 6.5

Tb 0.6 0.7 < 0.2 Zn 3.4

Ti 60 < 20 < 20

V 1.3 0.2 < 0.8

Zn 170 < 60 < 5

W 0.16 < 0.07 < 0.25


Post-mortem concentration changes of some trace elements

in lung specimen collection at different times during the autopsy.

(ng•g-1 wet tissue)

post-mortem sampling time

Element < 10 hours > 20 hours

Cesium 10 40

Iron 60000 204000

Rubidium 2830 5360

Selenium 120 155

Zinc 8200 13.500


Radio-labelled metabolic forms: blood collected after (2-7 days) intraperitoneal or intravenous injection of the radiotracers Storage : 1 month at – 20°C in polyethylene vials Freeze-drying : starting temperature –80°C ; final temperature 20°C.

Losses of Trace Elements during storage and free-drying of rat or rabbit radioactively-labelled blood

Loss (%) Loss (%) Radio-labelled element

Storage Freeze-drying

Radio-labelled element

Storage Freeze-drying 110 Ag (I) < 1 < 1 54 Mn (II) 0.07 <1 74 As (III) < 0,1 2.3 63 Ni (II) < 1 < 1 74 As betaine < 1 1.1 203 Pb (II) 0.8 0.9 74 As DMA 0.8 1.2 99 Mo (VI) 1.3 2.1 133 Ba (II) < 0.5 < 0.5 86 Rb (I) 0.1 1 106 Bi (III) 1.1 2 122 Sb (III) 2.1 1.8 109 Cd (II) < 0.1 <1 75 Se (IV) 0.09 1.5 141 Ce (III) <1 <1 75 Se metionine < 0.1 < 1 58 (Co) (II) < 0.08 < 0.5 113 Sn (IV) < 0.5 < 1 51 Cr (III) 0.2 1,5 182 Ta (V) < 1 < 1 51 Cr (VI) <1 1.1 202 Tl (I) < 2 1.2 152 Eu (III) < 0.3 < 0.1 48 V (V) 0.08 < 1 203 Hg (II) 2.8 3.8 185 W (VI) < 1 < 0.9 203 Hg DM 6.2 12 65 Zn (II) < 1 < 1


PRE-ANALYTICAL FACTORS: EXAMPLES FOR V, Se, Cr and As

V Se Cr As

PRE-ANALYTICAL

FACTOR

As Cr Se V

150 50 500 1000 ultraclean

room Laboratory Air

Quality (depletion factor)

ambient air

1 1 1 1

<0.1 <0.1 0.05 0.01 teflon

cannula SAMPLING

(ng/mL serum) syringe stainless steel needle

2.7 15.5 2.7 0.7

<0.08 <0.1 <0.05 <0.1

polyethylene CONTAINERS

(ng/mL) quartz

2.1 <0.5 <0.1 0.9

0.08 0.1 0.2 0.1 1 month

(- 20°C) STORAGE (loss%)

2 weeks (- 4°C)

1.3 0.6 0.3 0.17

0.6 0.1 1.1 3.5 from – 80°C FREEZE-DRYING

(loss%)

<0.5 3.6 4.5 13 400-500°C DRY- ASHING

(loss%)

Data obtained by radiorelease tests and by 74As, 51Cr, 75Se and 48V radiotracers.


TRACE ELEMENTS IN ANTICOAGULANTS

Range of elemental

concentration (ng·g-1)

Maximum estimated

contamination (ng·mL-1) a)

Typical concentration

in serum (ng·mL-1)

Element EDTA Heparin EDTA Heparin

Co 12 - 210 20 – 81 0.31 0.012 0.21

Cr 70 - 315 25 – 216 0.47 0.03 0.17

Cs 2 – 12 - 0.017 - 0.21

Mn - 610 – 1080 - 0.16 0.6

Rb 90 – 205 360 – 413 0.3 0.06 230

Sb 1.5 – 37 <1 0.06 - 0.5

Se 17 - 102 16 – 105 0.15 0.015 81

Zn 2455 - 18150 8790 - 95305 27.2 14.2 922 a) from the max. value and assuming that for 1 mL of blood 1.5 mg of EDTA and 0.15mg of heparin are used


Note: relative SD from 5 to 10 %

TRACE ELEMENTS IN FORMALIN μg.g-1

Br 0.07

Ce 13

Cr 0.21

Fe 3.5

Hg 0.01

I 0.07

K 60

Mg 1.6

Mn 0.09

Na 18

P 2.4

Rb 0.04

Sb 0.14

Se 0.03

Zn 6.2


Dissoluzione del campione

Attacco chimico convenzionale

HNO3 HCL HCLO4 H2SO4 H2O2 HF …..

Contenitore aperto

Bomba di dissoluzione

Forno microonde

SOLUZIONE FINALE

Controllo e valutazione : Contaminazioni

Perdite


TRACE ELEMENTS IN HIGH PURITY NITRIC ACID g L-1

Element ANALAR

(1995) ARISTAR sub.

(1995) Romil UpA

(2006) Human blood

ref. v. (ranges)

Al 18 <7 <0.020 0.3-7.5 As 10 < 2.8 < 0.015 0.4-10 Cd <1 <0.3 <0.002 0.1-1.5 Cr 130 4 <0.02 0.05- 0.4 Co 1.2 <0.25 <0.002 0.01 – 0.90 Cu 25 <3 <0.005 807 -1600 Hg <0.1 <0.04 <0.1 1.7 – 9.9 Mn 6.2 <3 <0.003 7-10 Ni 110 <3 <0.015 1-3-3.3 Se 0.04 <0.05 <0.015 76 -140 V 0.4 0.1 <0.015 0.09-0.75 Zn 17 <7 <0.08 4070-7500


Acid mixture chemical dissolution of biological matrix by different procedures. Insoluble fraction (%)detected by radiotracers experiments.

Sample: NIST-CRM “Peach Leaves”

HPTFB : (HNO3 , HF, HCl )160 C° - 8 h

Element Microwave Oven MWO

High Pressure Teflon Bomb

HPTFB

Open Teflon vessel OTV

insoluble f. % insoluble f. % insoluble f. %

Cr 8 < 0.5 3 Co 0.8 < 0.5 0.6 Sb 1.7 < 0.5 5 Sc 4.4 < 0.5 3.5 Ce < 0.2 < 0.2 < 0.2 Hf 36 3 40 Th 12 < 0.5 10 Zn < 0.5 < 0.5 < 0.5


The application of the Neutron Activation Analysis for trace metals determination in human brain tissues

and substantia nigra neuromelanin

In human brain many transition metals are necessary for important biological processes, and alterations in their homeostasis can produce neurotoxic effects. Toxicity pathways frequently involve oxidative stress through an increase of free radicals production (redox- reactions). The subsequent damages or death of cells can lead to neurodegenerative diseases, such as Parkinson’s Disease (PD) and Alzheimer Disease (AD)


Trace elements analysis in human brain tissues is an important step in understanding the PD pathogenesis. Thanks to a controlled separation procedure, NM was isolated from SN with no metal contamination. NM showed a great ability to bind large amounts of metals, especially Fe, suggesting that NM can play a protective role for brain neurons against the free toxic radicals deriving from redox active metal ions. Fe and Cu concentrations in brain tissues increase along with aging in normal subjects.


Trace metals and human health – Parkinson disease

Human substantia nigra in parkinsonian (A)Human substantia nigra in parkinsonian (A)and in normal subject (B)and in normal subject (B)

AA B


Substantia nigra pars compacta of normal (A) andSubstantia nigra pars compacta of normal (A) andparkinsonian subjects (B) parkinsonian subjects (B)

AA B


Brain tissue

Substantia Nigra collection, homogenization

1st wash. Milli-Q water , vortex shaking, sonication, centrifugation

Liquid phase: elimination of hydrosol. components

2nd wash. 3rd wash.

NaH PO , vortex, sonication, centrifugation

Liquid phase:elimination of big lipid molecules

4th wash. 5th wash. 6th wash.

SDS, TRIS, vortex,sonication, incubation, centrifugation

Liquid phase: elimination of proteins

7th wash. 8th wash.

NaCl, vortex, sonication, centrifugation

Liquid phase: elimination of SDS

9th wash. 10th wash. 11th wash.

Milli-Q water, vortex, sonication, centrifugation

Liquid phase:elimination of salts, hydrosol. res.

12th wash. Methanol, vortex, sonication, centrifugation

Liquid phase:elimination of water and some lipids associated to melanin

n-hexane, vortex, sonication, centrifugation 13th wash.

Liquid phase: elimination of other lipids associated to melanin

Dry under pure N

NEUROMELANINNIN

2 4

2

blanks analysis of the biochemical procedure used for Neuromelanin isolation from human brain tissues (SN).

Element Procedure with reagents alone

Procedure with synthetic melanin

Analysis of synthetic melanin

Total ng ng/mg ng/mg

Ag <1 <0.5 <0.5 As <0.5 < 0.3 <0.3 Au 0.05 0.03 0.05 Ba 29.9 <25 <25 Br 6.4 7.4 9.1 Ce 0.2 <0.5 <0.5 Co <0.05 0.01 0.01 Cr 0.3 1.5 1.8 Cs 0.11 <0.05 <0.05 Cu 2.21 30.4 32.2 Fe < 20 17.8 <15 Hf <0.01 <0.01 <0.01 Hg <0.5 <0.3 <0.3 I 4.8 7.5 8.7

La 0.17 0.1 0.1 Mn 2 4.6 5.3 Mo <0.5 <0.3 <0.3 Ni <100 <10 <10 Rb 0.88 0.45 <0.3 Sb 0.04 0.17 <0.1 Sc <0.05 <0.1 <0.1 Se <0.5 <0.5 <0.5 Th <0.08 <0.1 <0.1 V 0.32 0.8 0.5 Zn <5 5.2 <5

Note. Results are the mean of two independent analyses.


Trace metals in human brain tissues and their neuromelanines

Elemental blanks analysis of the biochemical procedure used for Neuromelanin isolation from human brain tissues (SN).

Element Procedure with

reagents alone Procedure with

synthetic melanin Analysis of

synthetic melanin

Total ng ng/mg ng/mg

Ag <1 <0.5 <0.5 As <0.5 < 0.3 <0.3 Au 0.05 0.03 0.05 Ba 29.9 <25 <25 Br 6.4 7.4 9.1 Ce 0.2 <0.5 <0.5 Co <0.05 0.01 0.01 Cr 0.3 1.5 1.8 Cs 0.11 <0.05 <0.05 Cu 2.21 30.4 32.2 Fe < 20 17.8 <15 Hf <0.01 <0.01 <0.01 Hg <0.5 <0.3 <0.3 I 4.8 7.5 8.7

La 0.17 0.1 0.1 Mn 2 4.6 5.3 Mo <0.5 <0.3 <0.3 Ni <100 <10 <10 Rb 0.88 0.45 <0.3 Sb 0.04 0.17 <0.1 Sc <0.05 <0.1 <0.1 Se <0.5 <0.5 <0.5 Th <0.08 <0.1 <0.1 V 0.32 0.8 0.5 Zn <5 5.2 <5

Note. Results are the mean of two independent analyses.


Elemental blanks of irradiation vials used in INAA for trace elements determination in human brain tissues and

Neuromelanin samples.

Element

Quartz vials Suprasil (0.6cm x

4cm height) Heraeus (Germany)

Polyethylene vials (1ml)

Kartell (Italy)

Total ng Total ng

Ag <1 <1 As <0.5 <0.5 Au 0.15 0.07 0.6 0.2 Ba n.d. <60 Br 1.5 0.8 11 0.4 Cd <10 <10 Ce <3 <3 Cl n.d. 870 ± 171 Co 0.3 0.05 <0.2 Cr 0.8 0.3 13 7 Cs 0.3 0.1 0.5 0.4 Cu n.d. <30 Fe 103 25 <850 Hf 0.13 – 0.1 <0.5 Hg <0.1 <0.1 I n.d. 5 2

La 0.8 0.5 <0.1 Mn n.d. 11 3 Mo <3 <3 Ni <20 31 8 Rb <1 2.8 1 Sb 10 6 <0.5 Sc 0.2 0.1 <0.5 Se <0.3 <0.3 Th 0.2 0.08 <0.1 U 0.4 0.2 <0.1 V n.d. 0.4 0.3 Zn 35 12 95 25

Note. Data obtained from 4 independent analyses; n .d. not determined


Elements in brain tissues and in their respective pigments (Neuromelanine) determined by Neutron Activation Analysis. Concentration in mg/kg (wet tissues).

Element Putamen NM from Putamen

Cortex NM from Cortex

Cerebellum NM from Cerebellum

Ag 0.05 7.23.6 0.11 20 8 0.02 6.7 1.9 As - 2.41.0 - 2.3 1.4 - 1.07 0.2 Au - 1.220.63 - 0.09 0.01 - 0.25 0.03 Ba 6.30 9869 0.71 - - 129 65 Co 0.03 0.820.04 0.02 0.82 0.15 0.02 0.4 0.2 Cr 0.39 193 0.11 47 27 0.19 8.9 0.5 Fe 833 4936203 274 1747 65 337 1031 499 Hg - 1.630.24 - 20 13 - 98 66 I 0.75 9.43.5 0.48 12.5 1.5 - 19 9 Mn 0.90 12.31.5 0.54 19 7 2.1 26 14 Mo - 8.82.8 - 5.8 1.1 - 2.6 1.4 Ni - 13837 - 96 1.1 0.52 30 20 Sb 0.01 0.680.21 - 0.53 0.07 - 0.54 0.22 Sc - 0.0230.002 0.01 0.03 0.01 - 0.018 0.010 Se 0.90 10.01.7 0.98 11.5 5.5 0.80 61 40 V 0.57 2.731.53 0.17 1.49 0.27 0.57 1.55 0.53 W - 4.140.32 - 6.64 0.22 - 1.79 0.97 Zn 80 47283 71 407 116 74 277 49

Biochemical Pharmacology, Aug 1;66(3), pp.489-94, (2003).

Pigment Cell Res., 17(6):610-7., Dec; (2004). INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia

Fe SN

LC

Age (years)

0 30 40 50 60 70 80 90 100

ng c

oppe

r / m

g w

et ti

ssue

0

10

20

30

40

50

60

70

80

Cu

LCLC LC

LC

Age (years)

0 10 20 30 40 50 60 70 80 90 100

ng c

oppe

r /

mg

wet

tis

sue

0

10

20

30

40

50

60

Cu

SN

LC

Proc Natl Acad Sci U S A.;101(26):9843-8. Epub Jun 21, (2004) INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia

Ti target Cyclotron irradiation

Ti target dissolution

H2SO4 /HF (Teflon PFA)

DRY to TiO2

FUSION (700 °C) Pt crucible

Na2 CO3 / Na2O2 50:1

2 H2O leaching

Centrifuge

Ti(OH)2 /TiO(OH)4 >99%

44m, 46, 47 Sc 100%

DISCARD (Bulk)activity)

48 V 90% in the surnatant

pH adj. to 3-4 with HCl

CHELEX 100

Ti (impurity)

48 V

eluition 20% NH3

48 V NCA Radiotracer

Chemical purity of 48V radiotracer solution. Ti and trace element concentration, determined by GF-AAS, ICP-ES and INAA

Element Analytical technique

Concentration

in ngg-1

Uncertainty % (1 SD)

Ti Et-AAS 810 11

Ti ICP-ES 935 9

As INAA 380 20

Au INAA 8.7 10

Co INAA 35 8.5

Cr INAA < 400 -

Cu ET-AAS 1.6 18

Cs INAA 120 9

Fe Et-AAS 970 18

Hg INAA 98 14

Sb INAA 9.6 11

Sc INAA 1.2 10

Zn INAA 115 12 Note: results obtained from three independent analyses

0

2

4

6

8

0 250 500 750 1000 1250 1500Energy (keV)

Counts(a.u.)

983.5

beta+

1312.0

944.4

928.3


Radiotracers in vivo experiments

Nuclear reaction : p, xnNat Ti 48 V

ARSENIC IN SHELLFISH Total As content in commercial frozen shrimps: mgkg-1 28 Assuming one meal of 100 g of shrimps = 2800 g total As Arseno-betaine > 90% of total As Inorganic As ingested about : 250 g ( no toxic exposure) Professional exposure ? Speciation is required !

Preparation of radio-labelled Arseno-betaine 73AsCl3 + LiCH3 73As(CH)3

Distillation + (ethyl bromoacetate)

73 AsCH3 + BrCH2CO2Et (ethyl bromoacetate)

(CH3)373AsCH2CO2Et

+ Br -

(As- betaine ethylester bromide)

(CH3)373AsCH2CO2Et

+ Br -

OH

-

(radio-labelled) (CH3)3

73AsCH2CO2- ARSENOBETAINE

TRACE ELEMENTS DETERMINATION IN BIOLOGICAL SPECIMENS AT NANOGRAM OR SUB-NANOGRAM

LEVELS

REQUIREMENTS

HIGH SENSITIVITY

- microsamples - ppb or fractions

PRECISION , ACCURACY

- “normal” levels - narrow window (essential./toxicity)

CHEMICAL METROLOGY

- protocol: methods, reproducibility, primary

standards, calibration, uncertainties...

- quality assurance: certified reference materials

DATA AND RELATIVE SD


I N R I M I N R I M

I N R I M

I N R I M

Toxic metals in the environment - K and P comparison of CCQM IAWG (2006)


I N R I M

I N R I M

I N R I M

As in shellfish

I N R I M


I N R I M

I N R I M

. CCQM-K45 and CCQM-P72 participants (2005/6)

INSTITUTE / ORGANIZATION COUNTRY

BAM

Federal Institute for Materials Research and Testing

Germany

CSIR - NML

CSIR - National Metrology Laboratory

Republic of South Africa

Hill Laboratories

New Zealand

IMGC

Institute of Metrology "G.Colonnetti"

Italy

INTI

Instituto Nacional De Tecnología Industrial

Argentina

KRISS Korea Research Institute of Standards and Science

Republic of South Korea

IRMM-SCK

Institute for Reference Materials and Measurements – SCK/CEN Belgian Nuclear Research Centre

European Union

LATU

Laboratorio Tecnológico del Uruguay

Uruguay

LGC United Kingdom

LNE Laboratoire National d’Essais

France

PTB Physikalisch-Technische Bundesanstalt

Germany

UME National Metrology Institute

Turkey

FOOD ANALYSIS : TRACE METALS DETERMINATION IN TOMATO PASTE

Participant

Digestion Method Instrumentation

LAB 5 Microwave digestion, HNO3/HCl/H2O2

IDMS Magnetic sector field ICP-MS

LAB 6 Microwave digestion, HNO3/HCl

IDMS Quadrupole ICP-MS with collision

cell

LAB 8 Microwave digestion, HNO3/HCl/H2O2

Multi-point calibration

Quadrupole ICP-MS with collision

cell

LAB 9 Dry ashing, HNO3/HCl/H2O


ETAAS, matrix modifier

NH4H2PO4

LAB 10 Microwave digestion, HNO3


ETAAS

LAB 11 HNO3/HCl Two point calibration

Quadrupole ICP-MS

LAB 12 Dry ashing, HNO3


GF AAS using THGA tubes,

matrix modifier Pd+Mg(NO3)2

Analytical techniques and procedures for Cadmiun determination

LAB 1

LAB 12

LAB 10

LAB 8LAB 6

LAB 11 LAB 5LAB 9

70

80

90

100

110

120

130M

as f

ract

ion

of C

d (

µg

kg-1

)


Reference Materials in life science

Agricultural – Nutritional related Materials

NI ST SRM 1567 Wheat Flour BCR CRM 063 Skim Milk Powder

NI ST SRM 1568 Rice Flour BCR CRM150 Skim Milk

NI ST SRM 1577 Bovine Liver BCR CRM151 Skim Milk

SRM SRM 1577a Bovine Liver BCR CRM 189 Whole meal Flour

SRM SRM 1566 Oyster Tissue BCR CRM 191 Brown Bread

SRM SRM 1570 Spinach BCR CRM 184 Bovine Muscle

BCR CRM 185 Pig Kidney

I AEA 359 Trace and Minor Elements in Cabbage Material

Human Health related Materials

NI ST SRM 2670 Human Urine (elevated) BCR 273 Single Cell Protein

NI ST SRM 2670 Human Urine (low)

I UPAC Freeze – Dried (Human Whole Blood) I AEA H-4 Animal Muscle

I AEA Animal Blood Freeze Dried

I AEA Mixed Human Diet H-9


Reference Materials in life science

I nstitution Material Elements analyzed SRM 1755 Pine Needles As, Cr, Hg, Se, Zn SRM 1572 Citrus Leaves As, Cr, Co, Hg, Se SRM 1573 Tomato Leaves As, Cr, Mn, Se, Zn SRM 1566 Oyster Tissue As, Cd, Cr, Hg, Se, Zn SRM 1577a Bovine Liver As, Cd, Cu, Hg, Se, Zn SRM 1567 Wheat Flour Hg, Se, Zn NI ST (NBS) SRM 1568 Rice Flour As, Cd, Co, Cu, Hg, Se, Zn SRM 1645 River Sediment Cd, Cu, Co, Mn, Hg, V, Zn SRM 1635 Coal (subbituminous) As, Cd, Cr, Cu, Mn, Se, V, Zn SRM 1632a Coal (bituminous) Cr, Cu, Hg, Mn, Se, Th,V SRM 1648 Urban particulate Matter As, Cd, Cr, Cu, Zn SRM 2783 Air Particulate Matter on Filter Media As, Co, Cr, Fe, K, Mn, Sb, V, Zn BCR CRM 060 Lagarosiphon m. Al, Cu, Hg, Mn, Zn BCR CRM 061 Platihypnidium r. Al, Cu, Hg, Mn, Zn BCR CRM 062 Olea Europea Al, Cu, Hg, Mn, Zn BCR CRM 274 Single Cell Protein Cu, Co, Mn, Se BCR CRM 150, 151 Skim Milk I BCR CRM 189 Wholemeal Flour Cu, Fe, Mn, Se, Zn, I RMM (BCR) BCR CRM 191 Brown Bread Cu, Fe, Mn, Zn BCR CRM 185 Bovine Liver As,Cd, Cu, Hg, Se, Zn, BCR CRM 186 Pig Kidney Cu, Fe, Hg, Se, Zn BCR CRM 180 Gas Coal As, Hg, Mn, Se, V, Zn BCR CRM 181 Coking Coal As, Hg, Mn, Se V, Zn BCR CRM 227 Estuarine Sed. As, Cr, Sc, Se, Zn BCR CRM 320 River Sediment As, Cr, Sc, Se, Zn I AEA Lichen 336 As, Br, Co, Cr, Sb, Se I AEA I AEA Cabbage 359 AS, Cr, Se I AEA NAT 7 Air Particulate Filter As, Co, Cu, Mn, Cr, Co, V, Zn,


with PM10 concentration more than 75 g/m3 …

Decreto Ministeriale n° 60 del 02/02/2002

Stage 6 (0,70 – 1,08 m)

Stage 7 (0,41 – 0,70 m)

Stage 8 (0 – 0,41 m)

Stage 5 (1,08 – 2,15 m)

Stage 4 (2,15 – 3,3 m)

Pre-impactor (> 9.0 m)

Stage 1 (6,0 – 9,0 m)

Stage 2 (4,6 – 6,0 m)

Stage 3 (3,3 – 4,6 m)

Air particulate matter collected onto filters

Stage 1 (6.0-9.0 m) Stage 5

(1.08-2.15 m)

Stage 8 (0-0.41 m)

Total Suspended

Particle (TSP)

NIST 2783 Air Particulate on Filter Media

Lab. 7: IMGC Lab.12: NIST

Reference Materials for life science : Trace elements in air particulate matter PM 2.5


NIST 2783 Air Particulate on Filter Media

Lab. 7: INRIM Lab.12: NIST

Reference Materials for life science : Trace elements in air particulate matter PM 2.5


Gaithersburg, MD 20899 Certificate Issue Date: 2 January 2002


METROLOGY ?

ENVIRONMENTAL

POLLUTION

Food Chain

ANALITICAL

CLINICAL CHEMISTRY

NUTRITION

DIAGNOSIS

DRUGS

HUMAN HEALTH

TERAPY

COSMETICS


YES !!

la scienza delle misure chimiche e qualità della vita mario gallorini i.n.r.i.m unità di...

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