Rapid and simple determination of selenium and other trace elements in very small blood samples by XRF

Selenium and other trace elements (Cu, Zn, Br, and Rb) were determined in very small (0.75 μL) human serum and mice whole blood samples, by an XRF method. Accurate results of elemental concentration were obtained without the need of exact volume measurement, because of the backscatter correction used. The XRF method is highly sensitive (M.D.L.=0.06, 0.13, 0.09, 0.07, and 0.05 ppm for Se, Cu, Zn, Br, and Rb, respectively), rapid (counting time—100 s/sample), easy to perform and therefore suitable for routine trace element analyses. The results obtained are in good agreement with the values reported in the literature.     

Interaction of trace elements in a longitudinal study of human milk from full-term and preterm mothers

Concentrations of 8 trace elements (Fe, Cu, Zn, Se, Br, Pb, Rb, and Sr) at different lactation time were measured by the PIXE multi-elemental technique. Time dependence and interelement correlations were studied. A total of 200 milk samples from 32 lactating mothers were supplied from 2 to 120 d after delivery of 26 full-term and 6 preterm infants. All elements showed a lognormal frequency-distribution. The Fe, Cu, Zn, and Se contents in preterm milk were found to be somewhat different with respect to full-term milk. Cu, Zn, Se, Br, Pb, and Rb concentrations declined with lactation time, both in pre- and full-term samples. Sr and Fe contents did not show any change with time. Detailed analysis of data by partial correlation and multiple regression methods was performed. No substantial differences between preterm and full-term samples were found in the results of partial correlation analysis. Cu and Zn were found to be correlated with lactation time, whereas the measured time dependence for the other elements has to be attributed to the effect of the existing interelement correlation. All the measured elements appeared to be correlated with at least one other element. In particular, Se was inversely correlated with Zn and directly with Cu. The zinc and copper contents in milk can therefore depend on the variation in the mother selenium intake.     

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 element correlations in the blood of indian women with breast cancer

Fourteen minor (Na, K, P, Fe) and trace (Br, Co, Cr, Cs, Hg, Rb, Sb, Se, Sr, Zn) elements have been determined in pre and postoperative blood samples of 10 breast-cancer-affected. Indian women and compared with controls. The study showed elevation of Cr, Hg, Fe, Rb, Sb, and Zn and lowering of Se, K, P, and Sr contents in the blood of cancer patients. Most elemental contents in pre and postoperative stages remain unaltered except Br, Co, and Sb. Statistical significance of Fe, Se, Zn, and Hg levels has been tested by box plots. Lowering of Se in blood (-54.4%) is correlated with its enhancement in cancerous breast tissue (94.7%) of various clinical stages. Se/Zn and Se/Fe ratios are lowered in the blood of cancer patients, whereas Na/K ratio is only marginally enhanced. An attempt has been made to correlate Se levels with the dietary intake and breast cancer risk vis-a-vis American and Japanese women.     

An elemental correlation study in cancerous and normal breast tissue with successive clinical stages by neutron activation analysis

Influence of trace elements in body metabolism and their physiological importance in various diseases have motivated their accurate and quantitative determination in biological tissues and fluids. Instrumental Neutron Activation Analysis (INAA) using short and long term irradiation has been employed to determine five minor elements (Cl, K, Na, Mg, P) and 15 trace elements (As, Br, Co, Cr, Cs, Cu, Fe, Hg, Mn, Rb, Sb, Se, Sc, Sr, and Zn) in cancerous and normal breast tissue from 30 patients of four clinical stages. Several elements show enhancement in cancerous breast tissue. Selenium shows maximum enhancement of 94.7% followed by K (81.6%), Sc (66.7%), Cu (58.2%), Na (48.5%), P (44.4%), and Zn (39.2%). Some element, such as Fe, Cr, and Mn, are depressed by 30.8, 30.1, and 12.8%, respectively. These elements compete for binding sites in the cell, change its enzymatic activity and exert direct or indirect action on the carcinogenic process accelerating the growth of tumors. This is further evidenced by histopathological examination of cancerous cells showing poor cytological differentiation. An attempt has been made to correlate trace element concentrations of Se, Cu, Zn, Rb, Br, Hg, As, Co, Fe, Cr, and Mn and the ratios of Se/Zn, K/P, Cu/Zn, Na/K, and Se/Fe with the clinical stages of cancer. Inhibition of enzymatic activity caused by variation in trace element concentrations results in immunological breakdown of the body system.     

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.     

Trace elements in hair of healthy children sampled by age and sex

Hair trace element (TE) (Cr, Mn, Fe, Zn, Cu, Br, Rb, Sr, Pb) levels from 336 healthy subjects were measured by the Proton-Induced X-ray Emission (PIXE) method. The subjects were divided in three groups: 157 full-term neonates (75 male and 82 female), 86 children (41 male and 45 female) ages 6 to 11 yr, and 93 adolescents (51 male and 42 female) 11 to 16 yr old. Cu, Zn, Cr, and Br show an increase from birth to 8 yr and then decrease. Fe, Mn, and Sr strongly decrease up to 8 yr and then remain almost stable. Sex differences are present in Fe, Zn, and Br of children and in Cu, Cr, and Br of adolescents.     

Influence of the coat color on the trace elemental status measured by particle-induced x-ray emission in horse hair

The influence of hair color on the trace elemental status in horse's hair has been studied. A current analytical technique such as particle-induced X-ray emission (PIXE) used in this study has provided reliable, rapid, easy, and relatively inexpensive diagnostic methods. Twenty-eight elements (Al, Br, Ca, Cl, Co, Cu, Cr, Fe, Ga, Hg, K, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, S, Se, Si, Sr, Ti, V, Y, and Zn) in mane hair were detected by the PIXE method. The gray hair contains significantly greter amounts of Cu, Ti, and Zn, and lower amounts of Br, Ca, Se, and Sr than those in other colored horse hairs (p<0.05). Those results measured in the horse's hair were similar to those found in human and dog hair. When interpreting a result, it should be kept in mind that hair color, especially gray hair, influences the concentrations of some elements in horse hair.     

A search for trace elements in some human intracranial tumors by instrumental neutron activation analysis

A investigation was undertaken to measure the presence of trace elements in some intracranial tumors using the instrumental neutron activation analysis technique. The following 20 minor and trace elements were investigated: Na, Mg, Al, P, Cl, K, Ca, Cr, Mn, Fe, Co, Cu, Zn, As, Se, Br, Rb, Sb, I, and Cs. Our results are compared with other trace element analyses in human brain tissue.     

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.     

Correlation between 25 element contents in mane hair in riding horses and atrioventricular block

The influence of atrioventricular block (AV-block) on the trace elemental status in a horse hair was studied. The particle-induced X-ray emission (PIXE) method has provided a reliable, rapid, easy, and relatively inexpensive diagnostic method. Twenty-five elements (Al, Br, Ca, Cl, Co, Cu, Cr, Fe, Hg, K, Mg, Mn, Na, Nb, Ni, P, Pb, Rb, S, Se, Si, Ti, Y, and Zn) in mane hair and serum were measured by the PIXE method. A horse hair with first- and second-degree AV-block contained significantly greater amounts of Br, Ca, Sr, and Zn than those of horses without electrocardiographic abnormalities, whereas there was no significant differences in the elemental contents of the serum of the both groups. Those results in contents of a horse hair suggest that the evaluation of the degree of ionic imbalance by this method might be used to predict the susceptibility of a horse to heart disease much before symptoms appear.     

X-ray fluorescence-based differentiation of neck tissues in a bovine model: Implications for potential intraoperative use

This study explores the possibility of using X-ray fluorescence (XRF)-based trace-element analysis for differentiation of various bovine neck tissues. It is motivated by the requirement for an intra-operative in-vivo method for identifying parathyroid glands, particularly beneficial in surgery in the central neck-compartment. Using a dedicated X-ray spectral analysis, we examined ex-vivo XRF spectra from various histologically verified fresh neck tissues from cow, which was chosen as the animal model; these tissues included fat, muscle, thyroid, parathyroid, lymph nodes, thymus and salivary gland. The data for six trace elements K, Fe, Zn, Br, Rb and I, provided the basis for tissue identification by using multi-parameter analysis of the recorded XRF spectra. It is shown that the combination of XRF signals from these elements is sufficient for a reliable tissue differentiation. The average total abundance of these trace elements was evaluated in each tissue type, including parathyroid and salivary gland for the first time. It is shown that some tissues can unequivocally be identified on the basis of the abundance of a single element, for example, iodine and zinc for the identification of thyroid gland and muscle, respectively.     

Multielement proton-induced X-ray emission analysis of betel leaves,betel nuts,and lime

Proton-induced X-ray emission (PIXE) method was employed to study the concentration of heavy elements in betel leaves, betel nuts, and mineral lime consumed in Bangladesh. The samples were collected from different parts of Bangladesh and analyzed by the thicktarget external beam technique of the PIXE method. The samples were exposed to the proton beam as 1-mm thick pellets and irradiated with 2.0-MeV protons having 20-nA beam intensity. The concentration of some 15 elements (K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, Rb, Sr, and Pb) was measured by comparison with a calibration curve constructed from the NBS orchard leaf standard SRM 1571. The validity of the procedure has been established by comparative measurements of Cu and Zn with atomic absorption spectrophotometry. The significance of the results is discussed in view of their implications in health and disease.     

Significance of serum trace element status in patients with rheumatic heart disease

It is known that certain trace elements can affect various heart diseases. In this study, we aimed to evaluate the changes in concentrations of certain serum trace elements in patients with chronic rheumatic heart disease (RHD). Serum analysis of selenium (Se), zinc (Zn), and copper (Cu) trace elements was assayed by atomic absorption spectrophotometry. RHD patients had significantly lower serum concentrations of Se and Zn than control subjects (p<0.05 and p<0.001, respectively). However, the serum Cu concentration was significantly higher in RHD patients than in controls (1.93±0.59 μg/L vs 1.06±0.29 μg/L; p<0.001). Similarly, the Cu/Zn ratio in RHD patients was higher than in control subjects (4.70±0.92 vs 1.68±0.45; p<0.001). Additionally, no significant correlation was found among these trace element concentrations and the functional capacity classes (p>0.05). RHD patients had decreased serum Se and Zn element concentrations and increased serum Cu element concentration. We suggest that Se and Zn deficiency might be contributory factors in the development of rheumatic heart disease, and a high Cu concentration and a high Cu/Zn ratio might reflect an ongoing inflammatory process in this disease.     

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.     

Effect of diabetes on trace-element concentration of blood

The effect of diabetes on trace elements concentration in blood of experimental animals has been studied by thin-target X-ray fluorescence analysis. Balb/C young adult mice, 6–8 wk old, were used in the study. About 100–200 μL venous blood was taken from each mouse for trace element analysis. The measurements were carried out on a commercial Wave-Length-Dispersive XRF System, with different X-ray tubes being used for maximizing the detection sensitivity of different groups of elements. Later on, the mice were made diabetic by an intravenous injection of Streptozotocin (250 mg/kg). Then, 2 and 3 wk after the injection, 100 μL of venous blood was drawn from each of the mice and analyzed for trace element concentration. In this way we were able to study the changes in blood trace elements caused by diabetes. The results and advantages of using experimental animals, under controlled conditions, to study trace element variations caused by different diseases, are discussed in the paper.     

Availability of Essential Elements in Nutrient Supplements Used as Antidiabetic Herbal Formulations

Five brands of antidiabetic herbal formulations as tablets, Diabetex, Divya Madhu Nashini, Jambrushila, Diabeticin, and Madhumeh Nashini, from different pharmacies were analyzed for six minor (Na, K, Ca, Cl, Mg, and P) and 20 trace (As, Ba, Br, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mn, Rb, Sb, Sc, Se, Sm, Th, V, and Zn) elements by thermal neutron irradiation followed by high-resolution gamma ray spectrometry. Further Ni, Cd, and Pb were determined by atomic absorption spectrophotometry. Most elements vary in a narrow range by a factor of 2–4 while a few others vary in a wide range, e.g., Na (0.05–0.67 mg/g), Mn (26.7–250 μg/g), and V (0.26–2.50 μg/g). All the five brands contain K, Cl, Mg, P, and Ca as minor constituents along with mean trace amounts of Cr (2.11 ± 0.67 μg/g), Cu (15.7 ± 7.11 μg/g), Fe (459 ± 171 μg/g), Mn (143 ± 23 μg/g), Se (238 ± 112 ng/g), and V (0.99 ± 0.93 μg/g). Jambrushila is enriched in Na, Ca, Mg, Cl, Fe, Cu, Se, and Zn, essential nutrients responsible for curing diabetes. Dietary intake of Mn, Fe, and Cu are greater than 10% of the recommended dietary allowance, whereas that for Zn and Se is less than 2%. Mean contents of toxic elements (As, Cd, Hg, and Pb) were found below permissible limits except in Jambrushila. Cr and Zn were inversely correlated with r = −0.81, whereas Rb and Cs exhibit linear correlation (r = 0.93) in five brands. C, H, N analysis showed C ∼ 55%, H ∼ 12%, and N ∼ 2% with a total of ∼70% organic matter. However, thermal decomposition studies at 700°C suggest less than 5% nonvolatile metal oxides. Herbal formulations contain minor and trace elements in bioavailable forms that favorably influence glucose tolerance and possibly increase the body’s ability to ameliorate development of diabetes.     

An XRF study of trace elements accumulation in kidneys of tumor-bearing mice after treatment withcis-DDP with and without selenite and selenocistamine

The effect ofcis-DDP treatment with and without selenite and selenocistamine was studied on kidneys of tumor-bearing mice. The amounts ofcis-DDP, selenite, and selenocistamine injected were chosen so as to be compatible with the treatment of humans. The animals were sacrificed at 7, 14, and 28 d after treatment. The kidneys were removed and subjected to trace element analysis by a novel X-ray fluorescence (XRF) method and for pathological assessment. The results show that following treatment withcis-DDP, K, Fe, Cu, and Zn reach a maximum level after 7 d; K, Fe, and Cu levels were significantly reduced by the addition of selenite. The level of Zn was reduced only in groups treated with selenite whereas that of K and Cu was reduced also in groups treated with selenocistamine with and withoutcis-DDP. The greatest increase in Pt and Se levels was reached 1 wk after injection withcis-DDP, with and without selenocompounds, and in the case of Pt was partly reduced by addition of selenite. Se returned to control values 2 wk after injection, although Pt was still high in all groups 2 and 4 wk after injection. The results corroborate the findings of our previous studies. The effect of selenocistamine incis-DDP treated mice was partly insufficient. The pathological examination of the kidneys did not show any differences in the effect of various additives during the study.     

Characterization of hypoglycemiant plants by total reflection X-ray fluorescence spectrometry

In this work, synchrotron radiation total reflection X-ray fluorescence spectrometry (SRTXRF) was used to determine trace elements in eight hypoglycemiant plants (Trigonella foenum graecum, Panax ginseng, Pfaffia paniculata, Myrcia speciosa, Zea mays, Harpagophytum procumbens, Syzygium jambolona, and Bauhinia forficate). The elements P, K, Ca, Ti, Mn, Fe, Cu, Zn, Rb, and Sr were detected in all medicinal plants investigated, whereas Si, S, Sc, V, Cr, Co, Ni, Se, Nb, Mo, Sn, Sb, Ba, Hg, and Pb were detected only in some of the samples. The concentration of elements in hypoglycemiant plants varied from 0.15 μg/g of Co to 3.0×10⁴ μg/g of K and the mean of experimental limit of detection for these elements were 0.14 and 3.6 μg/g, respectively.     

Direct analysis of human blood (mothers and newborns) by energy dispersive X-ray fluorescence

This work is an application of energy dispersive X-ray fluorescence (EDXRF) as analytical technique for trace element determination in human tissues. Potassium (K), calcium (Ca), iron (Fe), copper (Cu), zinc (Zn), bromine (Br), rubidium (Rb) and lead (Pb) were determined directly in blood samples from 66 mothers at delivery after full-term pregnancies. The corresponding 66 cord-blood samples of the newborns were also analysed, in order to find element correlations between maternal and newborn blood at birth. The studied samples were obtained from mothers aged between 15 and 39 years old, the gestational age being between 35 and 41 weeks and the newborns' weight between 2.310 and 4.310 kg. Samples were lyophilised and analysed without any chemical treatment. Very low levels of Pb were found both in maternal and fetal cord blood samples. Cu values ranged from 3 to 13 microg g-1, both for mothers and children. A correlation between Cu and Fe concentrations in maternal and fetal cord blood was found. Zn is considered as one of the key elements in newborn health. Concentrations between 10 and 40 microg g-1 were measured. A positive correlation between Br levels in mothers and children was observed. Positive correlations for mothers were observed between Zn and Rb as well as K and Fe. The corresponding correlations in fetal cord blood samples were not observed, however positive correlations were found between Ca and K; Cu and Fe. The mean concentrations for each element were similar in maternal and in fetal cord blood, except for Cu and Zn, being higher in maternal samples. No correlations between element concentrations and pathologies of the mothers were observed.