S.K. Sahoo 1 *, Z.S. Zunic 2, R. Kritsananuwat 1, H. Arae 1 and S. Mishra 1 1, Japan 2 Institute of Nuclear Sciences Vinca, Beograd, Serbia E-mail:sahoo@nirs.go.jp 3rd TRE-ICEP Veszprém-Hungary, May 16-18, 2012
Introduction In the past few years human hair and nail have been recognized as the invaluable tissues for monitoring human environmental exposure. The content of elements in human hair and nail is a good alternative indicator of the public and occupational exposure to natural radionuclides and other toxic metals over a long period of months or even years. Compared to other types of biopsy, hair and nail have different uses and even advantages over blood, urine or other accessible tissue. But in some regions, there is a conservative tradition or faith about hair and nail so it is difficult to collect hair or nail sample from that area.
Advantages of hair and nail Hair and nail have many of the characteristics of an ideal tissue for assays since they can be: Painlessly removed Easily collected and transported Highly stable at room temperature Normally discarded Contained normally relatively higher elemental concentration than other body fluids and tissues Reflected the actual exposure over a period of months, or even year. (Blood and urine tend to show current or recent body status)
Sampling location Niška Banja, a spa town of Serbia is located in the quaternary alluvium of River Nišava along the contact of the Koritnik limestone and travertine rock.
Sampling location The high value of 226 Ra concentration in soil and indooroutdoor radon concentrations found around Niška Banja make this region a high background radiation area. Radionuclide in soil Concentration (Bq/Kg) 226 Ra 10.2 750.2 232 Th 6.8-46.1 40 K 95.8-583.5 222 Rn Concentration Bq/m3 Indoor 6000 Outdoor 168
Sample collection 27 hair samples and 26 nail samples were collected from informed volunteers living in Niška Banja by cutting with ceramic equipment and stored in sealed plastic bags to avoid any contamination Transport to National Institute of Radiological Sciences (NIRS), Japan for chemical charaterization
Analytical procedures Hair and Nail samples Washed with ultra-pure water and acetone Hair samples Dry ashing 550 C 15 h Microwave Digestion 6 ml 68% HNO 3 and 1 ml 35% H 2 O 2 Nail Samples Acid digestion on a hot plate 6 ml 68% HNO 3 1 ml 35% H 2 O 2 U, Th and trace elemental analysis using ICP-MS and Rh as an internal standard
Advantages of ICP-MS Inductively coupled plasma-mass spectroscopy (ICP-MS) is the analytical technique of choice because of its: Multi-element capability Low detection limit High sensitivity Ability to measure a large range of concentration Simple experimental procedure
ICP-MS Plasma Frequency (MHz) RF power (kw) 27.12 1.30 HP-4500 Argon flow, l/min Plasma Auxillary Carrier Nebulizer Sampling distance, mm Sample uptake rate, ml/min 15.0 1.0 1.2 Babington type 6.0 0.4 Data acquisition Mode Number of point per peak Number of scan sweeps Dwell time per point, s Scan mass range, a.m.u. Peak jumping mode 3 100 0.3 103 238
Certified reference materials BCR-397 Human hair Cd, Hg, Pb, Se, Zn, As, Cu, Ni NIES-13 Human hair Cd, Cu, Hg, Pb, Sb, Se, Zn, Ag, Al, As, Ba, Ca, Co, Fe, Mg, Mn, Na, V JLK-1 Sediment Ag, As, Au, Ba, Be, Br, C, Cd, Cu, Cs, REEs, Th, U SRM 1547 Peach leaves Al, As, Ba, B, Cd, Ca, Cr, Cu, Pb, Mn, Ni, Sr, Th, U SRM 1570a Spinach leaves Al, As, B, Cd, Co, Cu, Pb, Mg, Mn, Ni, Sr, Th, U Use for validation of analytical procedure and accuracy evaluation. CRMs were digested by micorwave digestion. A good agreement in the measured values with the certificate was achieved.
Dry ashing - Microwave digestion Dry ashing - Microwave digestion Dry ashing - Microwave digestion Dry ashing - Microwave digestion Comparison of sample preparation 5 4 R² = 0,9952 Mn (µg/g) 6 5 R² = 0,9975 Sr (µg/g) 3 4 2 1 0 0 1 2 3 4 3 2 1 0 0 1 2 3 4 Open digestion Open digestion 0,10 0,08 0,06 R² = 0,9967 U (µg/g) 0,004 0,003 R² = -0,77 Cs (µg/g) 0,04 0,002 0,02 0,001 0,00 0,00 0,02 0,04 0,06 0,08 Open digestion 0,000 0,000 0,002 0,004 0,006 0,008 Open digestion
Elemental concentration in hair Element Present study (Niska Banja) Published * Mean S.D. Median range range Th 0.006 0.008 0.002 0.0002 0.0276 0.0003 2.7 U 0.017 0.018 0.013 0.0002 0.0771 0.005 1.28 Cd 0.032 0.048 0.010 0.0016 0.1508 0.008 16 Cs 0.001 0.001 0.001 0 0.0037 0.0002 2.4 Cu 15.560 9.637 17.576 0.468 33.116 0.3 294 Mn 0.678 0.952 0.365 0.035 3.885 0.03 50 Ni 0.437 0.416 0.280 0.037 2.193 0.002 28 Sr 2.406 2.523 1.536 0.110 8.322 0.14-860 * I. Rodushkin, M.D. Axelsson (2000) G. Samanta et al. (2004)
Elemental concentration in nail Element Present study (Niska Banja) Published * Mean S.D. Median range range Th 0.022 0.016 0.017 0.0023 0.0564 0.002 0.063 U 0.013 0.008 0.010 0.0025 0.0447 0.002 0.047 Cd 0.180 0.368 0.057 0.012 1.686 0.013 1.9 Cs 0.057 0.059 0.039 0.009 0.219 0.0008 0.0347 Cu 9.544 4.340 7.947 2.967 75.063 4.2 81 Mn 4.244 5.476 2.497 0.600 24.273 0.19 91.33 Ni 1.547 0.562 1.639 0.398 15.140 0.14 14.78 Sr 1.099 0.860 0.860 0.373 3.446 0.16 3.3 * I. Rodushkin, M.D. Axelsson (2000) G. Samanta et al. (2004)
Comparison of elemental concentration in hair from present study with reported values of healthy individuals from different countries Niska Banja South Serbia (1) Sweden (2) Italy (3) Ghana (4) Japan (5) New Zealand (6) USA (7) England (8) Norway (9) U (ng/g) Mn ( g/g) Ni ( g/g) Cu ( g/g) Sr ( g/g) Cd ( g/g) Cs (ng/g) 2-77.1 0.9-449 6-436 59 0.04-3.9 0.4-5.2 0.01-1.2 0.02-7.6 5.4-15 0.06-4.51 0.57-1.68 0.57-1.68 0.21-3.95 0.36 0.04-2.2 <0.01-4 0.11-1.6 0.03-10 1.3-72 0.17-3.0 1.62-4.52 0.4-1.3 0.44-7.10 0.5-33.1 7.4-32.9 8.5-96 0.29-280 0.7-12.8 6.0-69.1 3.42-8.12 3.42-8.12 4.6-19.4 76.5 0.1-8.3 0.2-5 0.14-5.54 0.001-0.15 0.012-3.63 0.01-0.36 0.03-1.72 4.2-6.8 0.05-0.57 0.36-1.51 0.10-0.43 0.11-0.99 0.041 0-3.7 1-13 0.2-1.9 0.78 1. S.K. Sahoo et al. (2012) 2. Rodushkin et al. (200) 3. Sonofonate et al. (2000) 4. Amartey et al. (2011) 5. Kamakura (1983) 6. Ward (1986) 7. ML (1989) 8. Ward (1988) 9. Gellein et al. (2008)
Data comparison with other populations The elemental concentration in hair and nail samples in the publications had a wide distribution range due to: geological differences Food habit method and detection limit of analysis The data obtained in our study on the high background radiation area are in agreement with previous publications dealing with other population.
Average concentration in hair Element Niska banja Range ( g/g) South Serbia DU area Range ( g/g) Th 0.0002 0.0276 - U 0.0002 0.0771 0.0009 0.499 Cd 0.0016 0.1508 0.012 3.63 Cs 0 0.0037 0.001 0.013 Han Pijesak Niska banja Cu 0.468 33.116 7.4 32.87 Mn 0.035 3.885 0.5 5.21 Ni 0.037 2.193 0.16 1.16 Gornja Stuba Bratoselce Sr 0.110 8.322 0.18 4.49 a high background radiation area an evidence of depleted Uranium area
Average concentration in hair Han Pijesak Niska banja Gornja Stuba Bratoselce
Sample Hair U Th H001 0.0098 0.0012 H002 0.0016 0.0190 H003 0.0112 0.0174 H004 0.0030 0.0014 H005 0.0371 0.0225 H006 0.0208 0.0276 H007 0.0771 0.0018 H008 0.0132 0.0099 H010 0.0083 0.0036 H012 0.0137 0.0014 H013 0.0181 0.0030 H014 0.0180 0.0014 H017 0.0162 0.0021 H018 0.0492 0.0006 H020 0.0091 0.0004 H021 0.0012 0.0002 H022 0.0049 0.0003 H024 0.0185 0.0031 H027 0.0127 0.0037 H029 0.0002 0.0003 H031 0.0043 0.0007 U and Th concentration in hair
Sample Nail U Th N001 0.0120 0.0191 N002 0.0201 0.0146 N003 0.0167 0.0196 N004 0.0236 0.0338 N005 0.0229 0.0083 N006 0.0318 0.0288 N007 0.0091 0.0085 N008 0.0096 0.0240 N010 0.0079 0.0023 N012 0.0080 0.0046 N013 0.0085 0.0121 N014 0.0046 0.0118 N017 0.0093 0.0072 N018 0.0079 0.0166 N020 0.0314 0.0107 N021 0.0077 0.0564 N022 0.0088 0.0373 N024 0.0105 0.0440 N027 0.0151 0.0539 N029 0.0044 0.0080 N031 0.0097 0.0346 U and Th concentration in nail
Elemental concentration in hair and nail
Elemental concentration in hair and nail
Considering the column charts, elemental in hair and nail can be grouped into 3 groups as follow: 1. Concentration in Hair > Nail : Cu, Sr, U Cu is absorbed by Keratin in hair very well 2. Concentration in Hair < Nail : Cd, Mn, Ni, Th 3. Concentration in Hair <<< Nail : Cs
Conclusion ICP-MS is a powerful tool for multi-element analysis of hair and nail samples that can provide elemental coverage and low detection limits. Hair and nail samples can be as the good biological material for worldwide environmental monitoring. The main problem of using hair and nails is the external or surface contamination and lack of standardized methodology.