Application of PIXE analysis to the study of the regional distribution of trace elements in normal human brain

A particle-induced X-ray emission (PIXE) analysis method is presented, which allows measurement of eight elements (i.e., K, Ca, Mn, Fe, Cu, Zn, Se, and Rb) in human brain samples of only a few mg dry weight. The precision and accuracy of the method were investigated by analyzing animal brain matter with both PIXE and instrumental neutron activation analysis (INAA). The method was applied to measure the 8 elements in 46 different regions of 3 human brains. The sections analyzed originated from either the left or the right cerebral hemisphere, brain stem, and cerebellum. For one of the brains, sections were also analyzed from 26 corresponding regions of both hemispheres. For all elements, similar concentrations were found in the corresponding areas of the left and right sides of the brain. The concentrations (in μg/g dry weight) of the elements K, Fe, Cu, Zn, Se, and Rb were consistently higher in cortical structures than in white matter. Deep nuclei and brain stem, which have a mixed composition, showed intermediate values for K, Zn, Se, and Rb. A hierarchical cluster analysis indicated that the various brain regions clustered into two large groups, one comprising gray and mixed matter regions and the other, white and mixed matter brain areas.     

Proton-induced X-ray emission analysis of atherosclerotic plaques of the carotid bifurcation

The trace elements of both calcified atherosclerotic plaques and plaque-free vessel walls of the carotid bifurcation from 31 autopsies were investigated using the proton-induced X-ray emission (PIXE) method. The trace elements studied were phosphorus (P), calcium (Ca), chrome (Cr), iron (Fe), copper (Cu), zinc (Zn), lead (Pb), selenium (Se), bromine (Br), strontium (Sr), and rubidium (Rb). All samples contained Fe and Zn. Mercury (Hg) was not detected in any of the samples studied. All plaque-free samples contained Cu and almost all Br and Ca, none Sr. All calcified atherosclerotic plaques contained Ca and almost all Br and Sr. The relative levels of Ca were higher in the calcified plaques than in the plaque-free vessel walls. The relative value of Ca in calcified and uncalcified samples was greatest in the group who had died because of cardiovascular disorders and smallest in the group who had died from other causes. There was a strong positive correlation between the Ca and Sr of the plaque samples and between the P and Br of the plaque-free samples.     

Trace elemental analysis of cancer-afflicted intestine by PIXE technique

Trace elemental analysis was carried out in the biological samples of cancer-afflicted intestine using the particle-induced X-ray emission technique (PIXE). A 2-MeV proton beam was employed to excite the samples. From the present results, it can be seen that the concentration of the elements Cr, Fe, and Ni are higher in the cancerous tissue of the intestine than those observed in the normal tissue, whereas the concentration levels of the element Zn is slightly lower in the cancer tissue of intestine than that observed in the normal tissue. The concentrations of S, Cl, K, Ca, Ti, Mn, Co, and Cu in the cancer tissue of the intestine are in agreement with those observed in the normal tissues within standard deviations. The present results support the previous observations that Ni and Cr are carcinogenic agents. The observed slightly low levels of zinc in the cancer tissue of the intestine suggest that zinc could possibly inhibit the tumor growth and development of neoplastic transformation. For correctly assessing the role played by the trace elements in initiating, promoting, or inhibiting cancer in various organs, there is a need for the acquisition of more data by trace elemental analysis from several investigations of this type undertaken in different regions.     

Trace-element analysis of human teeth and bone by proton-induced X-ray emission

The objective of our study is to correlate Fe, Cu, Zn, Pb, Br, Sr, and Mo concentrations in human teeth crown, root, and bone. The samples, teeth from a lower jaw bone and the lower jaw bone itself, were obtained from a 97-yr-old female cadaver. Two hundred milligrams of crown and root of the teeth, and lower jaw bones were fixed to a Kapton film and irradiated with a 3.8-MeV external proton beam from an 8-MV FN Tandem Van de Graaff Accelerator. TheF-test was used to analyze the difference between crown, root, and bone. It resulted in the following: For Fe, Zn, and Pb, the comparison showed significant differences among the three sample populations of tooth crown, root, and bone; for Cu, Br, Sr, and Mo, the comparison showed no significant differences among all three sample populations. Several elements in the samples were highly interrelated.     

Elemental analysis of blood of nigerian hypertensive subjects

Proton-induced X-ray emission (PIXE) has been used to obtain the concentrations of 11 elements (P, S, Cl, K, Ca, Fe, Cu, Zn, Br, Rb, and Cd) in whole-blood samples of 16 hypertensive subjects (mean age: 52.5 ± 0.5 yr) and 18 age-matched controls (mean age: 51.5 ± 0.5 yr) in a Nigerian population. The results of the study indicate that the hypertensive subjects have significantly higher mean concentration of Cl, Cd, Cu, and Zn when compared with the controls, and the mean concentration of P, K, and Ca was found to be significantly lower in the hypertensive group in comparison to the controls. Furthermore, the Zn : Cd ratio was found to be significantly higher in the controls than in the hypertensives, and the Cu : Zn ratio was significantly higher in the hypertensives.     

Distribution of elements in rabbit liver lobule

The elemental composition of rabbit liver was determined by the PIXE and micro-PIXE methods. The mean concentrations of P, S, Cl, K, Fe, Cu, Zn, and Rb measured by both methods were similar. The latter method also allowed for localization of elements within lobule territory. It has been found that some elements are more prevalent in the veins (Cl, Fe) and others in the liver parenchyma (P, Cu, Zn). Moreover, Zn showed the characteristic intralobular distribution. Some methodological aspects of microbeam application to biological materials were also discussed.     

Quantitative trace element analysis of human nails with external beam PIXE

External beam PIXE (Particle Induced X-ray Emission) analysis with a proton beam of 2.4 MeV was used to study trace element concentrations in human nails. The suitability of PIXE analysis regarding nail samples without any pretreatment besides washing was investigated. The main emphasis has been on the ability to obtain absolute concentration values and a new accurate method for nail sample standardization has been developed. Concentration values for the elements Ca, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, and Pb were determined from human nail samples. A comparison was made with nail samples taken from different fingers and toes to monitor intraindividual variation, and nails of different healthy individuals to get a view of the interindividual differences. The concentrations were also measured in relation to time in order to observe any possible short-term changes. The results are compared with the previous studies reported in the literature. The nail analysis is also compared to hair analysis in terms of detection limits, number of elements determinable, and standardization of the results.     

Matrix effects in PIXE analysis of aerosols and ashes

A comparison of instrumental neutron activation analysis (INAA) and proton-induced X-ray emission (PIXE) results for sizefractionated atmospheric aerosols (“coarse” and “fine” fractions with an equivalent aerodynamic diameter of 2–10 Μm and < 2 Μm, respectively, or the PM10 fraction) showed that PIXE yielded significantly lower results for the PM10 and coarse fractions, especially for elements with a low Z resulting from a particle size effect. Somewhat lower PIXE results were also obtained for the fine fraction of atmospheric aerosols. A correction is also needed for irregularly shaped deposits of combustion aerosols collected by a cascade impactor in 11 size fractions ranging from 0.016 to 14.3 Μm, as well as for thick samples of fly and bottom ashes. An equivalent layer thickness (ELT) model is proposed to correct the matrix effects in PIXE. The approaches for the calculation of ELT using a comparison of PIXE and INAA results or by comparing PIXE results obtained using two different incident proton beam energies (1.31 and 2.35 MeV) are described. The correction for the ash pellets and irregular deposits are also discussed.     

A high resolution PIXE measurement for blood plasma ultrafiltrate

A new technique for investigation of elemental concentrations in subfractions of blood plasma is presented. The method is composed of the ultrafiltration of plasma in the presence of ethylenediaminetetraacetic acid (EDTA) and the measurement of the elemental composition by proton-induced X-ray emission (PIXE). The blood samples were collected from both healthy persons and patients suffering from breast cancer. The main emphasis in this study was on the determination of loosely bound copper (Cu) in plasma subfractions containing substances with molecular mass under 10,000, but zinc (Zn) and iron (Fe) contents of these fractions were also determined. The detection limits obtained with this method for Cu, Zn and Fe were ≈10 ppb (wet wt).     

Use of rubidium, manganese, and zinc as tracers to measure intestinal permeability by PIXE analysis

Intestinal permeability has been suggested to be closely linked with the etiology or activity of Crohn’s disease. However, current methods for measurement of intestinal permeability are too laborious for routine examination, as they require urine collection and/or use of radioisotopes. The present study was performed to develop a more convenient and safer method for assessing intestinal permeability using blood samples rather than urine. Rats with indomethacin-induced enteritis were orally administered Rb, Mn, and Zn as tracers. Intestinal permeability was determined by assaying the levels of Rb, Mn, and Zn in blood samples by particle-induced X-ray emission (PIXE). The distributions of Rb, Mn, and Zn in the small intestine after administration were analyzed by micro-PIXE. The conventional PIXE analysis showed that the levels of Rb and Zn in the blood in the enteritis group were correlated with the grade of enteritis. The micro-PIXE analysis showed that Rb, Mn, and Zn were translocated into the wall of the proximal small intestine 5 min after administration, and this effect was more conspicuous in the enteritis group than in controls. Analysis of blood or small intestine tissue samples using the PIXE allows determination of both intestinal permeability and the route of permeation.     

Study of trace elements in blood of cancer patients by proton-induced X-ray emission (PIXE) analysis

Proton-induced X-ray emission (PIXE) analysis was employed to determine the concentrations of six elements in the plasma and erythrocytes of 18 cancer subjects (15 males and 3 females) with neoplastic disorders and in 70 controls (35 males and 35 females). It was found that the concentrations of Br, K, and Zn were significantly elevated in the erythrocytes of the cancer subjects compared to the controls, whereas the concentration of Fe was significantly depressed, but with no difference observed in the concentration of Ca. In the plasma, the concentrations of Br, Cu, Ca, and K were significantly elevated, whereas the concentrations of Fe and Zn were found to be significantly depressed compared to the controls.     

Comparison between levels of trace elements in normal and cancer inoculated mice by XRF and PIXE

Determination of Rb, Br, Se, Zn, Cu, Fe, and Br/Rb ratio in tissues of mice inoculated with colon and melanoma cancer cells is described. A group of 19 Balb/c mice inoculated with C26 colon carcinoma, 4 C57B1/6 mice inoculated with B16 melanoma, and 13 control mice of both kinds were under investigation. The study was conducted on samples of blood, liver, kidneys, colon, and skin, and the trace element levels in normal and inoculated mice were compared. The inoculation was by subcutaneous injection either at the back or intrafootpad. The blood samples were taken 1, 2, and 3 wk after inoculation, and after 4 wk all the animals were sacrificed. Two nondestructive, complementary analytical methods were used: a modified X-ray fluorescence (XRF) for solid tissue and particleinduced X-ray emission (PIXE) for blood samples. The detection limit (DL) in the PIXE method was 0.35 (μg/g dry wt in 600 s counting time and in XRF, 1 μg/g dry sample for Rb, Br, Se and Zn and 2 μg/g for Cu and Fe in 200 s counting time. In all the cases studied, cancerous tissue developed at the site of the injection, and a significant difference in the trace element levels was observed between tissue samples obtained from normal and inoculated mice. The most pronounced effect was an increase in Rb level in the tumor by a factor ranging between 4 and 10 relative to normal tissue, with a corresponding decrease in the Br/Rb ratio (p < 0.05). Smaller changes were found in the Br, Se, Zn, and K levels. The changes in trace element levels in the inner organs were much smaller and seem to be influenced by the site of injection.     

Cadmium,copper, and zinc levels in the rice and rice field soil of Houston,Texas

Fifty-one pairs of hulled rice samples and the soil from which each rice sample was grown were analyzed for heavy metals in August, 1979, in order to estimate the background contamination of cadmium (Cd), copper (Cu), and zinc (Zn) in rice grown in the Houston, Texas area. Both samples were divided into three groups by soil types and colors. The cadmium concentration in Texas rice was only one-half to one-quarter lower than that of Asian rice. However, the levels of Cu and Zn in rice in Texas were similar to those reported. Soil heavy metals were lower than ever reported, but these values were consistent with the geochemical characteristics of the Texas Houston area. No particular relationship was found between the three metals in rice and the metals in soil where the sampled rice was grown.     

Trace-element analysis by charged-particle-induced reactions as a tool for physicians and chemists

The object of this article is to apprise physicians and chemists of nuclear analytical techniques and, in particular, of ion beam analysis (PIXE and PIGE) for the purpose of application to the clinical diagnostic method. The feasibility of the technique, sampling, and sample preparation for trace element analysis in biological and biomedical samples has been described previously (1–3). Analysis data from normal human blood samples and biomedical samples by ion beam reactions have been compared at the end. Emphasis will be placed on the use of the analytical technique on determination of the range of trace and toxic elements in human blood samples.     

Effect of chewing betel nut on measurements of salivary progesterone and estradiol

The measurement of steroids in saliva is both simple and non-invasive and has been widely used in field and clinical-based research. The observance of particular cultural practices by some populations, however, may hamper accurate hormonal analyses. The present study evaluated the effects of one such practice-the chewing of betel nut-on the accurate measurement of salivary progesterone and estradiol. A time series experiment was conducted among Bangladeshi women who are regular users of betel nut. Salivary steroids were analyzed by radioimmunoassay in samples collected prior to and then 30, 60, 120, and 240 min following betel quid use. Results show no significant difference between basal steroid levels and those obtained 60, 120, and 240 min after chewing betel nut. We conclude that with specific collection protocols that take into account time since chewing, salivary steroid analyses can be undertaken in populations among whom the practice of chewing betel nut is endemic.     

Some observations on the trace element concentrations in human dental enamel

The concentration of trace elements has been measured for dental enamel from 86 healthy human teeth using particle-induced X-ray emission (PIXE). The majority of the teeth (n = 70) were collected from dentists in the county of Oxfordshire in the United Kingdom, although a smaller group (n = 16) were collected from Cornwall. The elements K, Ca, Mn, Fe, Co, Ni, Cu, Zn, Sr, Pb, and Hg have been detected and statistically analyzed by grouping according to sex, age, and geographical location. The concentrations of Fe and Cu were found to be lower in the teeth from female donors (P < 5%) and are believed to result from the continued burden of blood loss during menstruation. Strong positive correlations (P < 0.1%) were found between Ca, Co, Ni, and Zn for all groups; these elements were also found to exhibit a negative correlation (P < 1%) with age for teeth from female donors. This is believed to be related to decalcification during the menopause. Pb was found to exhibit a positive correlation (P < 5%) with age for both sexes, and is believed to substitute for Ca in the Ca hydroxy apatite (HAP) within the dental enamel.     

Determination of eight elements in six human cancer cell lines and two human normal cell lines by PIXE

Gastric, hepatic, and pulmonary cancer cell lines, and the third passage of normal gastric and pulmonary cell lines were analyzed by proton induced X-ray emission (PIXE) method. The contents of element Sr, Ca, Fe, Zn, P, K, Cu, and As in the cell lines were determined. The Sr, Ca, Fe, Zn, and As contents in cancer cell lines were significantly lower than those in the normal cell lines (p less than 0.05), whereas there were no significant differences for the P, K, and Cu contents (p greater than 0.1). The results suggest that the need of some essential elements has been diminished in cancer cell proliferation.     

Effects of diluted sewage on biochemical parameters of carp,Cyprinus carpio

Changes in blood parameters and plasma elements of carp exposed for 14 days to diluted sewage were studied. The levels of elements in small amounts of plasma samples were also determined by PIXE analysis. Carp exposed to diluted sewage increased ammonia, glucose, Mg, Cu and Br, and decreased protein, K, Fe and Zn in plasma. It is concluded that sewage exposure causes considerable stress responses even with sufficient DO in water. PIXE for multielement analysis was also found to be quite useful even for small amounts of samples.     

Application of a proton microprobe to biomedical research

The capability of PIXE analysis to simultaneously detect trace elements with Z≥14, with a high power of detection, can be exploited in biomedical research if the diameter of the proton beam is reduced to micrometer dimensions. In this case, trace analyses of small particles or small parts of a larger specimen are rendered possible without deteriorating the detection limits of PIXE. The measurements yield a completely new type of information on the biological microstructure. In order to fully utilize the abilities of the combined method, however, sample preparation techniques, and irradiation procedures have to be adapted to each analysis problem. Examples of application of the Bochum Proton Microprobe will be used to demonstrate how and to what extent this can be achieved for different types of biomedical problems.     

Reference values for the concentrations of As,Cd, Co,Cr, Cu,Fe, I,Hg, Mn,Mo, Ni,Pb, Se,and Zn in selected human tissues and body fluids

This report attempts to formulate reference ranges of elemental concentrations for 15 trace elements in selected human tissues and body fluids. A set of samples consisting of whole blood, blood serum, urine, milk, liver, and hair were chosen and considered for 15 elements of biological significance: As, Cd, Co, Cr, Cu, F, Fe, I, Hg, Mn, Mo, Ni, Pb, Se, and Zn. The results represent wholly or partially data received from 40 countries of the global regions of Africa, Asia, Europe, North, South, and Central America, Australia, and New Zealand. This survey, even if qualitative, has been useful in demonstrating certain trends of trace-element scenarios around the world. It is of course recognized that both diet and environment exert a strong influence on the distribution pattern of several elements, such as As, Cd, Mn, Pb, Se, and Zn. A limited comparison of the available information on soil status of different countries reflected some interesting associations for elements, such as Mn and Zn. Importantly, this study revealed that only a few countries were in a position to identify a reasonable amount of data on samples requested for this project. Regretably, for a number of countries, any dependable data for even such essential elements as Cu, Fe, and Zn were not available. In view of the nutritional importance of many elements, the time is ripe for concerted efforts by intergovernmental agencies to initiate investigations or commission task forces/projects to generate reliable reference data for selected global regions, which sadly lack data of any kind at present.