ienza delle misure chimiche e qualità della Mario Gallorini I.N.R.I.M Unità di Radiochimica e Spettroscopia c/o Università degli Studi di Pavia Torino - 25 Gennaio 2007
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
EUROTERVIT - JRC- Ispra 1990-2000
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
(radio- release 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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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.
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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)
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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.
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
Trace metals and human health – Parkinson disease
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
Human substantia nigra in parkinsonian (A)Human substantia nigra in parkinsonian (A)and in normal subject (B)and in normal subject (B)
AA B
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
Substantia nigra pars compacta of normal (A) andSubstantia nigra pars compacta of normal (A) andparkinsonian subjects (B) parkinsonian subjects (B)
AA B
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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.
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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.
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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)
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
I N R I M
I N R I M
I N R I M
As in shellfish
I N R I M
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
Multi-point calibration
ETAAS, matrix modifier
NH4H2PO4
LAB 10 Microwave digestion, HNO3
Multi-point calibration
ETAAS
LAB 11 HNO3/HCl Two point calibration
Quadrupole ICP-MS
LAB 12 Dry ashing, HNO3
Multi-point calibration
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
)
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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,
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
Gaithersburg, MD 20899 Certificate Issue Date: 2 January 2002
INRIM Unit of Radiochemistry and Spectroscopy c/o University of Pavia
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
YES !!