International Journal of PIXE

Lead (Pb), cadmium (Cd) and uranium (U) present no known biological function but are toxic in various concentration ranges. Pb and Cd lead generally to nephrotoxicity consisting in proximal renal tubular dysfunction and accumulation while U has been reported to induce chemical kidney toxicity, functional and histological damages being as well mainly observed in proximal tubule cells. This work address the question of Cd, Pb, and U cytotoxicity, intracellular accumulation and repartition after acute intoxication of renal proximal tubule epithelial cells. After cells exposure to different concentrations of metals for various times, morphological changes were observed and intracellular concentrations and distributions of toxic metals were specified by PIXE coupled to RBS. Cell viability, measured by biochemical tests, was used as toxicity indicator. A direct correlation between cytotoxicity and intracellular accumulation in renal epithelial cells have been established. Finally, intracellular Pb and U localizations were detected while Cd was found to be uniformly distributed in renal cells.

Half of the freshwater fish fauna in New Zealand is diadromous (i.e., migrates between the sea and freshwater); unimpeded access to and from the sea is important for the conservation of diadromous fish. The genus Galaxias (Galaxiidae: Osmeriformes) contains five diadromous species that spawn in freshwater, migrate to sea as larvae, and then migrate back into freshwater as 50-55-mm juveniles. Microchemistry of the otolith, a calcified structure in the fish's head, allows an independent test of assumptions about these migrations. Concentric layers of CaCO₃ with some Sr as SrCO₃ comprise the otolith, creating a chronosequence that reflects a fish's migratory history. More Sr accumulates in the otolith when a fish is in seawater than when it is in freshwater. High-resolution nuclear microscopy was used to measure the molar ratios of Sr/Ca in two galaxiid species. Otoliths of inanga (G. maculatus) caught in freshwater all showed a central zone of 100-200 μm in radius with Sr/Ca of 0.008-0.012, indicating early rearing in the sea. Sr/Ca values decreased to 0.001-0.002 as the fish moved into freshwater. Of six adult koaro (G. brevipinnis) caught in a river with sea access and no lakes, five had migrated to sea but one had not, raising questions about the generalized assumptions of migration.

In this study we compared the structure and trace elements contents of three kinds of hair samples from Mexican infants: pre-Columbian mummies, mummies from the relatively recent past (less than 200 years) and from a healthy living 6-year old boy. External and internal morphology was characterized by electron microscopy. Trace elements were determined by PIXE (broad proton beam and microprobe method). The three groups showed differences in structure and elemental content. Calcium-containing mineral inclusions were observed in mummy hair but not in modern hair. Also, a progressive enrichment with age was observed for elements such as Ca, Fe, Mn, and Pb, indicating a postmortem incorporation. Other elements, such as Zn, Cu and Sr, less affected by post-mortem processes, were present at very similar levels, but still in a higher proportion relative to modern hair, preventing any possible relation to the individual's health.

We developed a micron-CT consisting of micro-beam system and X-ray CCD camera (Hamamatsu photonics C8800X), whose element size is 8μm×8μm and a total number of image elements 1000×1000 gives an image size of 8mm×8mm. The sample is placed in a tube of a small diameter, which is rotated by a stepping motor. The transmission data through the sample are taken with characteristic Ti-K-X-rays (4.558 keV) produced by 3MeV proton and α particle micro-beams. After image reconstruction using an iteration method, 3D-images of small objects namely, hair and small ants were obtained with a spatial resolution of ~5μm. It is expected that our micron-CT can provide cross sectional images of in-vivo cellular samples with high resolution and can be applied to a wide range of research in biology and medicine.

This contribution deals with the morphological and elemental characterisation of biomedical samples with high energy (MeV) focused ion beams with special emphasis on high spatial resolution in the sub-micrometer regime and very low minimum detection limits (< 50 μM/l) in trace element analysis. This paper gives an overview of recent applications at the LIPSION laboratory using PIXE (Particle Induced X-ray Emission) and STIM (Scanning Ion Transmission Microscopy) as well as STIM-Tomography.

The prostate is an accessory sex organ that secretes essential components of semen. High levels of Zn are found both in the prostate and its secretion. In the present study, the effects of environmental stress on zinc levels in the prostate were examined in Wistar male rats exposed to tributyltin chloride (TBTC), an environmental toxicant, by inductively coupled argon plasma–mass spectrometry (ICP-MS) for determination of Zn concentration in lobe-dissectioned specimens and micro-PIXE analysis for determination of site-specific distribution of Zn in prostatic sections. A single oral administration of TBTC to rats at a dose of 90 μmol/kg resulted 14 days later in a decrease in the Zn level in the dorsolateral prostate but not the ventral prostate. The TBTC did not affect the weight of the prostate tissue. Micro-PIXE imaging found Zn both in the lumen and the epithelium of the lateral prostatic tubules. PIXE spot analysis revealed that the epithelium of the control rat contained levels of Zn 4–fold higher than in the lumen and that Zn in the epithelium decreased selectively after TBTC exposure.

The giant kokopu, Galaxias argenteus, is a native New Zealand freshwater fish which optionally undergoes a marine larval phase after hatching near a river mouth. The marine phase is indicated by a high Sr:Ca ratio in the core of the otolith of the adult fish. Elemental scanning for Sr and Ca with a proton microprobe is one of the most useful techniques for identifying and quantifying the presence of the marine core. However, many individuals, whether or not they show the marine core, exhibit one or more concentric rings of slightly elevated Sr levels outside the core. Examples of this behaviour are given and quantitative estimates of the degree of elevation of the Sr levels are made from both area and line scans using high efficiency detectors for Sr detection. It is suggested that these Sr variations reflect adult fish migration within a freshwater habitat, perhaps seeking a brackish zone in which to spawn.

Particle Induced X-ray Emission (PIXE) technique has been used to determine the trace elements present in fourteen representative human gallstone samples collected from eastern region (Orissa) and thirteen representative samples collected from southern region (Chennai) of India. PIXE irradiation of the samples has been carried out by using the 3 MV tandem type horizontal pelletron accelerator facility at Institute of Physics, Bhubaneswar with proton beam of energy 3 MeV. In the present investigation, twenty one trace elements like S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Br, Rb, Sr, Y, Zr, Mo and Pb have been estimated in all the three types of gallstones viz. cholesterol stone, mixed stone and pigment stone. While sulphur in cholesterol stones in the eastern region was less than that of the southern region, sulphur was present as a minor element in the pigment stones of both the regions. Less concentration of copper in the gallstones from eastern region is another interesting observation. The lower values of copper in the patients of eastern region may be due to different types of food habits. The concentrations of all the elements in the southern region pigment stones have higher values than that of the eastern region. Moreover, the concentrations of Fe and Mo in cholesterol stone and pigment stone samples in southern region have also higher values than in eastern region. The current PIXE study is of its first kind in this eastern region of India.

In order to explore the distribution and the loss way of inorganic substances in bone and to provide scientific basis for prevention and therapy of osteoporosis, Proton Induced X-ray Emission (PIXE) method is used for the determination of elemental concentration in femoral head from five autopsies and seven patients with femoral neck broken, and Synchrotron Radiation X-ray Fluorescence (SRXRF) microprobe analysis technique is used to scan a slice of the femoral head from its periphery to its center, via cartilage, compact and spongy zones. The results show that the concentrations of P, Ca, Fe, Cu, Sr in a control group are higher than those in a patient group, but the concentrations of S, K, Zn, Mn are not significantly different. The quantitative computerized tomography of elemental distribution, such as Ca, P, K, Fe, Zn, Sr and Pb in bone slice tissue including cartilage, substantial compact and substantial spongy, is investigated. Combined with the correlations between P, K, Zn, Sr and Ca, the loss way of minerals and the physiological functions of some metal elements in bone are also discussed.

With the aim of characterizing polymer-based drug delivery systems a combination of Scanning MeV ³He microbeam Nuclear Reaction, Backscattering and Particle Induced X-ray Emission (PIXE) techniques has been developed. This, together with gravimetric and UV techniques has been applied to characterize both water infusion and drug effusion for three in-mouth polymer–drug systems. Preliminary results are presented from the exposure of polymers, containing drug at a level of 9% by weight of the dry polymer, to both pure water and a phosphate buffered saline solution at 37°C.

We have measured fluorine concentration in samples taken from humans, such as sweat, saliva, snivel and tear in addition to hair, nail and urine, together with those of other elements by means of a three-detector measuring system. Fluorine has been known as one of the most toxic elements as well as arsenic, lead, mercury and cadmium which contaminate environment over the wide area. This system allowed us to discuss about fluorine concentration related to other elements'. A standard-free method established by us has been applied to quantitative analyses of these samples. By means of these methods, quantitative analysis becomes possible for samples of less than 1mg or 1μl and it is quite suited to tear, snivel and sweat. As one of the conclusion, sweat, snivel and tear give useful information in addition to the samples which have been traditionally analyzed. It is found that titanium concentration in a body is well estimated by analyses of sweat, snivel and saliva, arsenic concentration can be estimated by sweat analysis in addition to urine analysis, and nickel is well evaluated by sweat and tear. In this way, it is required to estimate elemental concentration in a human body by measuring various clinical samples. Especially, correlation between fluorine and arsenic is found in a few bio-medical samples.

Metal monitoring is one of the major concern of biological and bio-medical research. The Proton Induced X-ray Emission (PIXE) which is a multi elemental and non-destructive technique for trace element analysis has been set up at the Folded Tandem Ion accelerator (FOTIA) at BARC, Trombay, Mumbai, India. A few of the bioscience applications carried out at the Van de Graaff Laboratory using PIXE will be summarized in this presentation. Tea leaves obtained from the eastern region of India were analyzed by PIXE using 2.5 MeV protons. Trace elements such K, Ca, Ti, Cr, Ni, Mn, Fe, Cu, Zn, Rb and Sr were present in varying concentration in the tea leaves. The soil analysis showed Ti, Y, Zr and Nb in addition to the elements shown in the leaves. Blood samples of children exposed to vehicular traffic exhaust were analyzed for its Pb content and it was seen that the concentrations of Pb in the blood of the children were high (6.0 μg/ml). Pb poisoning of air due to vehicular exhaust is a major concern and cause for air pollution leading to high blood lead levels in children.

The release of metals and other elements from dental and orthopedic implants into the tissues has been a matter of numerous studies. However, the conditions that favor such a phenomenon have not been completely understood so far. PIXE microbeam technique has been applied to study the spatial distribution of the elements released from the implants. Subjects of interest have been implants made from commercially pure titanium, Ti-6Al-4V alloy and 316 LWM surgical steel. Block sections of five dental implants, as well as tissues surrounding one metal spine plate and two hip prostheses, have been examined. Rabbit bones containing copper inclusions have been taken for microbeam analysis to complete the study about the metal release processes. Samples from the tissues and bone slices have been analyzed using PIXE microbeam setup at National Laboratories of Legnaro, Italy. Traces of the implant metals, including titanium but with the exception of vanadium, were found in the surrounding tissues and maps of their elemental distribution at different distances from the contact surface with implants have been obtained. The origin and the differences of the release rate of the observed metals have been discussed.

A prototype of micron-CT for biological research is being developed at Tohoku University. This micron-CT uses a point X-ray source with a spot size of 1μm and an X-ray CCD with 1000×1000 pixels of 8μm×8μm, achieving a spatial resolutions of the order of micro-meter. The event data obtained by the X-ray CCD is statistically poor and the 3 dimensional filtered back projection (3D FBP) algorithm, generally used in image reconstruction of X-ray CT, is not suitable because it is highly sensitive to statistical noise. Hence, we applied the expectation maximization (EM) algorithm for image reconstruction and developed an image reconstruction method using 3D EM algorithm. To confirm the validity of the reconstruction method, we irradiated two hairs inside a micro tube and reconstructed the CT image applying both EM and FBP algorithm on projection data. With 200×200×200 voxels of 4μm×4μm×4μm in the field of view, the computation time was less than 2 mins per iteration on a DELL Precision 650 Workstation 3.2GHz. The resulting EM image showed a better contrast than FBP image, and in the EM reconstructed CT image, we were able to reconstruct the micro tube of 270μm diameter and 45μm wall thickness and to visualize the two hairs inside.

Balance of essential elements in organisms might be changed by environmental stresses. Small fresh water fish, Medaka, was burdened with X-ray irradiation (total dose: 17Gy), keeping in salty water (70% NaCl of sea water) and keeping in metal containing water (10ppm of Cr and Co). These stresses are not lethal doses. Essential elements in liver, gall bladder, kidney, spleen, heart and brain in the stress-loaded fish were measured by PIXE method and compared with a control fish to determine the effect of the stresses. Various changes of the elemental contents were observed. Effect of X-ray irradiation was the smallest among the stresses. Relatively high content elements such as P, S, Cl and K were hardly affected with the stresses examined in this work. The effect of Cr on the metal balance seems to be larger than the other stresses. As PIXE method can analyze many elements in a small sample simultaneously, change of elemental distribution in small organisms induced by environmental stresses can be determined readily.

In order to shed light on the long-range transport of atmospheric pollutants in the Northeast Asian regions, we studied the chemical components in rime and fresh surface snow on Mt. Moriyoshi (altitude: 1454 m), located on the Sea of Japan side of northern Honshu, Japan, near Northwest China and Southwest Russia. Rime and fresh surface snow samples were collected at Juhyou-Daira (near the summit; altitude: 1200 m) in February 2004. The pH range of rime samples was 4.2 – 4.9, and the electric conductivity (EC) range was 52 – 282 μS/cm. On the other hand, the pH range of snow samples was 4.5 – 4.9, and the EC range was 17 – 77 μS/cm. The elemental compositions, ionic species and particle shapes of these samples were determined and/or observed by PIXE, ion chromatography and SEM-EDX analysis. PIXE analysis of the rime and snow samples revealed 24 elements, of which Na, Mg, Al, Si, K, Ca, Ti and Fe were found to be the major components. Comparing the determined values of rime and snow sample elements, rime samples were several times to several dozen times higher than snow samples for almost all the elements. For determined values of ionic species, rime samples were several times to several dozen times higher than snow samples. With the aid of SEM-EDX analysis, many small silicon-rich spherical particles were observed in the rime samples. Small silicon-rich spherical particles were also found in the snow samples. The existent forms of chemical components in rime will be an important factor when we consider the origin of air pollutants transported over long distances in the Northeast Asian regions.

The target-chemetherpy directed by the radiation was tested by the two methods; 1) facilitate the carboplatin; and 2) the capsules that is decomposed by irradiation IN VITRO. The human leukemic cell (OCI/M2) cultured in RPMI 1640 was treated by the 0.5 Gy of ⁶⁰Co γ ray after the 4μmol of carboplatin administration. The uptake of carboplatin expressed as Pt signal was measured by the micro PIXE camera. The increased uptake of carboplatin was observed in the cell. There was also an increased uptake of Cu. The chelating of Cu resulted in the maximum uptake of carboplatin under the combined treatment of radiation and carboplatin. As for the capsules, it was prepared by polymerizing the alginate by CaCl₂, whose core was carboplatin. The 0.5 Gy irradiation was performed to the capsules. The carboplatin's emission was observed using a micro PIXE camera. As a result, the carboplatin was emitted from capsules, which was almost 10 %. Those two methods (increased uptake of carboplatin by low dose irradiation and the capsules decomposed by irradiation) is considered to be useful for the better therapeutic gain and less adverse effect.

At certain locations in the Wellington Region, pollution episodes due to air particulate matter are known to occur from time to time. Traditional gravimetric analysis of airborne particulate matter is unable to provide information on the sources contributing to air particulate concentrations. Ion Beam Analysis (IBA) is one of the few non-destructive techniques that can be used to identify the elemental composition of air particulate matter on a filter sample. In this work IBA was used to characterise air particulate matter in two size fractions, PM_20. and PM_10-2.0, collected at a monitoring station in Masterton, New Zealand. Elements with atomic mass above neon were measured by the PIXE technique. Elemental carbon was measured with a light reflectance device. Elemental 'fingerprints' of contributing sources were determined by performing factor analysis of the elemental composition. The results indicate that 'Sea Salt' and 'Soil' sources are major contributors to the coarse (PM_10-2.0) fraction and 'Combustion' sources dominate the fine (PM_2.0) fraction of air particulate matter. Analysis of seasonal differences was a useful tool in elucidating source profiles.

Since 1991 ANSTO's ion beam analysis (IBA) laboratory has been sampling fine atmospheric particles every Wednesday and Sunday at urban and rural sites in New South Wales. Multi-elemental accelerator-based IBA techniques were used to characterise major components and significant trace elements with minimum detectable limits close to 1 ng/m³. Observed mass concentrations will be compared with air quality US EPA standards and proposed Australian fine particle NEPM guidelines. Trace elements strongly associated with source fingerprints responsible for high air pollution will also be discussed in this paper.

An enhanced sample preparation method for PIXE analysis is described allowing to separate and concentrate arsenic ions of different oxidation states in water samples. Arsenate ions are separated from arsenite ions by co-precipitating into 10 ppm indium hydroxide colloids that are generated at pH 4.0 in a 25 ml solution containing 1 ppm phosphate ions and 25 ppm sulfate ions. Arsenite ions are oxidized to the pentavalent state with permanganate ions and adsorbed by indium hydroxide colloids generated afterwards in solution. The standard procedure for collecting the colloids adsorbing arsenic ions on Nuclepore filter of 0.2 μm pores is based on an investigation of the pH-dependence of the recovery of dissolved arsenic ions and the obtained standard calibration curve covers the concentration range from 1 to 100 ppb for arsenic ions. The prepared targets were examined for 5 to 10 minutes by 3 MeV proton beam (0.7-4 nA beam currents). The lower detection limit of arsenic in a 25 ml aquatic sample is 0.3 ppb for the arsenic-precipitated targets based on the 3σ error of background counts integrated over the FWHM of arsenic peak in the PIXE spectrum. This sample preparation technique was then applied to analyze concentrations and oxidation states of arsenic in a river basin where hot springs are located upstream being possible sources for releasing arsenic in the river.

PIXE and Ion Beam Analysis are one of the few techniques that can be used to identify the elemental composition of air particulates without destroying the filter sample. They are key tools for identifying the sources and determining the relative contribution of biogenic and anthropogenic sources of air particulate matter pollution in our environment. Over the last 8 years, specialised equipment has been designed and built at the New Zealand Ion Beam Analysis facility in Lower Hutt for semi automated analysis of air filters. The equipment and experimental techniques have been refined to improve sensitivities for many of the elements in the periodic table. At GNS, sensitivities have recently been further improved by using two X-ray detectors simultaneously with different amounts of X-ray filtering and collimation. The average limit of detection is improved from 66 ng/cm² (typical for a setup using a single detector) to 35 ng/cm² using two detectors simultaneously. The New Zealand Ion Beam Analysis facility now routinely analyses air particulate matter collected on filters from several locations around New Zealand. In this paper, results of air particulate studies from several locations in the Wellington region are presented.

A microbeam system has been developed for the analysis of single aerosol particles. Combination of PIXE, RBS and off-axis STIM methods enabled simultaneous analysis for hydrogen to metal elements. Aerosol particles were collected on thin polycarbonate film (~0.3 μm) resulting in good signal-to-noise ratio. Quantitative elemental correlation was measured for single aerosol particles. A total of 270 particles were analyzed and clustered into 4 groups. The analysis system reveals the chemical composition of aerosol particles and is a powerful tool for source identification.

Fine particulate matter PM_2.5 samples have been regularly collected in the Mexico City Metropolitan Area commencing in the year 2003, when a new PM_2.5 sampling network was made fully operational. Samples have been collected during 24 h periods every third day using US EPA Federal Reference Method (FRM) samplers. In this study, the elemental composition of two sets of PM_2.5 samples collected at the "Merced" sampling site located in downtown Mexico City (19°25'28''N, 99°07'09''W) were determined by PIXE. Ten elements (S, Cl, K, Ca, Ti, Mn, Fe, Cu, Zn, and Pb) were detected in most of the samples. Some others appeared occasionally, but only those consistently detected were considered significant for the analysis. Other analytical equipments operating simultaneously at the site provided complementary data of several variables such as SO₂, NO₂, NOx and CO. An enhanced database was thus built adding these results to the database derived from previous PIXE analysis. Further statistical analysis of the database allowed identifying four principal sources in two seasons and permitted to estimate the corresponding source apportionment.

The International Atomic Energy Agency (IAEA) via the Regional Co-operative Agreement (RCA) has identified air particulate matter pollution as a major transboundary environmental issue in the Australasian region. Sixteen countries in the region spanning from Pakistan to the Philippines and from China to New Zealand are participating in the regional programme "RAS/7/013– Improved information of urban air quality management in the RCA region" that started in 1997. New Zealand is the lead-country for this programme in which nuclear analytical techniques, such as particle induced X-ray emission (PIXE), neutron activation analysis (NAA) and X-ray fluorescence spectrometry (XRF) are used to measure key elements in PM_2.5-0 and PM_10-2.5 filters from GENT stacked samplers collected twice weekly. Major sources of air particulate matter pollution are identified using statistical source apportionment techniques. To identify transboundary air particulate matter pollution events, the data is collated in a large database. Additionally, the data is used by end-users of the participating countries in the programme. An overview is presented.

Environmental samples (rice, soil, river soil and water, running water, pine needle, milk and grass) are collected from the area surrounding Kaya Memorial Takizawa Laboratory (radioisotope waste management laboratory) neighboring Nishina Memorial Cyclotron Center for detection of radionuclides in them. Rice, which is the most important diet for Japanese, was prepared using a nitric acid ashing and analyzed using ordinary vacuum PIXE for detection of the elemental concentration. Comparison of potassium-40 between the detected value using Ge radiation detector and the analyzed value using PIXE was carried out for accuracy. We examine the change of elemental concentration in rice samples harvested in the different rice paddies in 2001 and in the same rice paddy during 13-year period. There is little change in those.

We have measured concentration of multi elements including fluorine in mushrooms collected in Iwate prefecture, Japan. As a result, it was confirmed that the sample preparation method, which was established for the plant samples, is applicable to the mushroom samples and the elemental concentration was obtained in good accuracy and reproducibility. Moreover, it becomes clear that the mushroom takes a specific element selectively depending on its species, and it is found that to analyze various kinds of mushrooms is of great use as a way of monitoring environmental pollution. Fluorine concentration is found to be 7-68 ppm in the mushroom samples. However, clear correlation of fluorine and other elements was not observed in each sample in this study.

PIXE is a trace analytical method. During the sample preparation process for PIXE analysis, there may be loss of element due to sublimation or evaporation that must be considered for quantitative analysis. To avoid such problems, suitable droplet PIXE system has been developed for determination of elemental abundances of liquid samples in atmospheric condition. The system has several dripping modes in order to enhance advantage of dripping method over static liquid target method. The droplet-PIXE system has been applied to a few case studies. Its advantages over other PIXE system will be discussed in this paper.

Coal ash is a major component of solid residue resulting by coal-fired thermal power plants. Trace elements like Mn, Cu, Zn, As, Se, Mo and Pb in fly ash and pond ash samples collected from ten coal-fired thermal power plants in India, and crops (rice, wheat, maize grain and straw and onion bulbs) grown on coal ash treated soils at Angul, Bakreswar and Farakka sites of India have been analysed at Institute of Physics by Particle Induced X-ray Emission and Energy Dispersive X-Ray Fluorescence techniques. Our analysis shows that the concentrations of trace elements of grains are higher than in straws. Increasing trends in the uptake of micro-nutrients by the crops due to application of coal ash, were recorded, but within permissible limits.

A new analytical method based upon the combination of Pulsed UV (PUV) photolysis and detection of elements via X-ray fluorescence (PIXE and XRF) techniques has been developed and shown capable of rapid and reproducible trace element analyses with increased sensitivity in solid and liquid foods. Pulsed UV-induced photolysis allows for a rapid, acid-free digestion of food samples. Thin or thick samples are then prepared for analyses with X-Ray Fluorescence methods (PIXE and XRF). The combined method was optimized for analysis of Co, Cr, Cu, Fe, Pb, Tl, Bi and Zn, and validated with NIST food standards. The analytical sensitivity of the PUV/PIXE/XRF method allows the study of selected sections of fruits and vegetables, a database useful in determining the uptake, transport mechanisms, and the distribution of trace elements in fresh produce. When applied to edible parts, the analytical results shall also provide valuable information to ascertain potential human health risks due to the presence and bioaccumulation of toxic elements in foods. Practical applications include field studies for analyses of fresh produce as well as for evaluating environmental impact derived from current agricultural practices. Chemical profiling of agricultural crops and soils for improved management and increased productivity using the precision agriculture approach is also being explored.

PIXE has been used to extract the elemental composition of cobalt blue glazes in 55 dated and undated terracotta sculptures from the Florentine della Robbia school. The data, consistently with previous work, show a rapid change in the composition of blue glazes and locate it just before 1520. The change is marked essentially by the presence of arsenic and bismuth in the glaze and by reduced amounts of iron and nickel after 1520. We speculate that this is due to procedures introduced to industrialise the blue pigments production more than to exploitation of different families of cobalt minerals. We observe in this unique ensemble more variability of products than was known before.

Coins are important archaeological objects and analysis of metallic compositions of coins can provide valuable information regarding coin minting, methodology, classification, provenance, art, culture and economics of the minting time. In-air Proton Induced X-ray emission (PIXE) is a good scientific method for non-destructive analysis of coins preserved in museum. Selected ten Kushana copper coins (3^rd-4^th century A. D.) from the Orissa State Museum, Bhubaneswar have been analysed, for the first time as preliminary work, by using an external beam PIXE facility at Institute of Physics. Our study reveals that copper is the main constituent of the Kushana coins, with minor/trace elements like titanium, iron, and nickel, zinc, lead and bismuth.

Fossil fragments of bone and teeth that are exposed to a humid environment take up fluorine from the surrounding soil and accumulate it in their mineral phase. In cortical parts of long bone diaphysis a fluorine concentration profile can be observed, which carries information on the exposure duration of the buried object in its shape. The distribution of fluorine in a sample however is strongly influenced by environmentally induced processes of bone diagenesis, i.e. alteration in the structure and composition of bone mineral and degradation of the organic components that may make the time information indistinct.

PIGE (Proton Induced Gamma-ray Emission) is a precise and fast analytical technique to determine the quantitative fluorine content and its distribution in cross sections of bone and tooth specimen non-destructively. The simultaneous detection of Ca by PIXE (Proton Induced X-ray Emission) provides additional information on the sample topography. Cracks, alteration haloes and the porosity, which is typical for human bone samples, are parameters which have direct influence on the fluorine uptake and transport during burial. This contribution outlines the combined approach of using PIGE and PIXE measurement to understand some aspects of the complex environmental impact that impedes exposure age dating by fluorine diffusion profiling.

A group of 15 ink drawings from the Genoese painter Luca Cambiaso (1527-1585), who is considered the most prolific and amongst the most innovative drawers in the history of arts has been analysed with PIXE that is the most convenient technique for the instrumental investigation of inks. The metallo-gallic nature of the inks has allowed singling out some key elements that are used to differentiate the inks within the group of artefacts and extend the stylistic analysis with objective data. Infrared reflectography has been used for the overall examination of drawings. Even in the limited group selected for this first study it seems possible to confirm and strengthen the distinction between originals and school copies and to raise some questions about the washing technique and the use of mixed (carbonaceous + metallo-gallic) inks.

Paintings from Baghcheh Joogh castle belonging to the Qajar era have been analyzed using the PIXE method. The pigments of over 16 samples from various parts of castle have been analyzed in the present study. A proton beam of 2 MeV energy generated by a Van de Graaff accelerator was used for the analysis. A Si(Li) detector detected the resulting X-ray spectra. The PIXE elemental analysis was expected to determine the details of golden themes of the wall paintings.