Blood-copper and zinc levels and consequences of cardiovascular complications: a study by INAA and FAAS

To understand the role of Cu and Zn in human blood both in controls as well as in cardiovascular (CVD) patients, whole blood samples of 181 CVD and 185 controls between the ages of 20–66 years were investigated. Instrumental neutron activation analysis (INAA) and flame atomic absorption spectrophotometric techniques were successfully employed to quantification Cu and Zn levels. The mean blood-Cu levels (1.50 mg L⁻¹) were found as enhanced whereas Zn levels (5.88 mg L ⁻¹) were reduced in cardiovascular patients group as compared to 0.90 and 6.70 mg L⁻¹ for Cu and Zn respectively in controls. Cu/Zn ratios for CVD patients are also higher than in control subjects. Negative correlation exists between Cu and Zn levels in both controls and patient groups. However, when the CVD patients were checked for their systolic and diastolic pressure it was found that copper concentrations in these patients was significantly increased (p < 0.001) with the rise of blood systolic pressure so a positive correlation was observed between copper and systolic pressure. Zn on the other hand has an inverse relation with systolic as well as diastolic pressure (p < 0.001). Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, and triglyceride (TG) in blood samples have also been determined and their probable role in the CVD complication has been observed. A positive correlation of blood-Cu with TC, TG, and LDL-C indicates that rise in blood-Cu levels may initiate the development of CVD. An increase in Cu/Zn ratio can instigate the cardiovascular risk factor. The findings from this study can definitely update our knowledge of the role of Cu and Zn in the development of CVD risk in humans.     

INAA and flame AAS of various vegetable reference materials

Summary INAA and flame AAS have been used for the analysis of a large number of vegetable reference materials. Out of all determined elements (28 by INAA and 11 by AAS) nine are common for both methods (Ca, Cd, Co, Cu, Fe, Mg, Mn, Na, Zn) and for these the possibilities of the two methods have been compared.     

Instrumental neutron activation analysis of the hair of metallurgical workers

Hair samples were collected from 20 metallurgical workers (10 males and 10 females) and from 59 control subjects (32 males and 27 females), whose jobs do not indicate a specific occupational exposure. The concentrations of ten minor and trace elements (Al, Co, Cu, Fe, Mg, Mn, Sb, Se, V and Zn) were determined by instrumental neutron activation analysis (INAA). The statistical data distributions, the sex and age influences in these elemental concentrations and the average values obtained for the control group were compared with published data. The effect of occupational exposure to the metallic elements was reflected in elemental composition of hair by significant higher concentration levels of Al, Co, Cu, Fe, Mg, Mn, Sb, V and Zn in the hair of the exposed group, when compared with the control group.     

The complementary use of radioanalytical methods and trace analysis methods for the characterisation of environmental IAEA reference materials

The IAEA laboratories have access to both nuclear analytical methods and conventional instrumental methods for trace analysis for the characterisation of its environmental and biological reference materials. A technical concept integrating the homogeneity data, the recommended test portion mass and the uncertainty on the trace element concentrations is explored using a combination of INAA, XRF, ICP-MS, ICP-AES, and AAS. The above approach is illustrated by selected examples of reference materials currently under evaluation for their trace element content. They include some or all of the following elements Al, As, Br, Ca, Cd, Co, Cr, Cs, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Rb, Se, Sr, Th, U, Zn in environmental biomonitor organisms (lichen, algae) and air dust filters.     

Preparation and characterization of a set of IAEA reference air filters for quality control in air-pollution studies

Several sets of reference air filters were prepared as part of an IAEA evaluation of the performance of laboratories involved in air-pollution studies. Each set comprised three polycarbonate membrane filters, two of which were loaded with urban air particulate matter (APM) obtained in Vienna or Prague, and one unloaded filter. The filters were loaded by filtration of a suspension of the APM materials in water. The homogeneity both of bulk APM materials and of the loaded filters was evaluated and found suitable by determining several elements by instrumental neutron-activation analysis (INAA), proton-induced X-ray emission (PIXE), and micro-X-ray energy-dispersive fluorescence analysis (micro-EDXRF). After evaluation of the homogeneity, INAA, PIXE, EDXRF, atomic absorption spectrometry (AAS), inductively coupled plasma optical emission spectrometry (ICP-OES), and ICP mass spectrometry (ICP-MS) were used to characterize the filter materials and establish "target values" and their associated standard deviations for 15 elements. Problems encountered during the preparation of these unique, simulated air filters and the criteria for setting both the target values and standard deviations are presented.     

Preparation and characterization of a set of IAEA reference air filters for quality control in air-pollution studies

Several sets of reference air filters were prepared as part of an IAEA evaluation of the performance of laboratories involved in air-pollution studies. Each set comprised three polycarbonate membrane filters, two of which were loaded with urban air particulate matter (APM) obtained in Vienna or Prague, and one unloaded filter. The filters were loaded by filtration of a suspension of the APM materials in water. The homogeneity both of bulk APM materials and of the loaded filters was evaluated and found suitable by determining several elements by instrumental neutron-activation analysis (INAA), proton-induced X-ray emission (PIXE), and micro-X-ray energy-dispersive fluorescence analysis (μ-EDXRF). After evaluation of the homogeneity, INAA, PIXE, EDXRF, atomic absorption spectrometry (AAS), inductively coupled plasma optical emission spectrometry (ICP–OES), and ICP mass spectrometry (ICP–MS) were used to characterize the filter materials and establish “target values” and their associated standard deviations for 15 elements. Problems encountered during the preparation of these unique, simulated air filters and the criteria for setting both the target values and standard deviations are presented.     

INAA of elemental contents in fingernails of esophageal cancer patients

Fingernails of pathologically diagnosed normal people, light esophageal epitheliosis patients, severe esophageal epitheliosis patients and esophageal cancer patients were irradiated and their elemental contents were determined by INAA. Multivariate statistical treatment of Ca, Cl, K. Mg, Se and Zn data shows that esophageal cancer patients are distinguishable from non-cancer patients. The accuracy computed by neural networks is greater than 80%.     

Nutritional and Environmental Properties of Algal Products Used in Healthy Diet by INAA and ICP-AES

Spirulina platensis alga sampled in the Caribbean Sea and seven other commercial algal products available on the Italian market of different origin and aspect, have been analyzed by instrumental neutron activation analysis (INAA) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). By neutron irradiation and -ray spectrometry (INAA), as many as 20 elements could be measured instrumentally without any chemical treatment. Cu, Mg, Mn and Pb were determined after dissolution of the sample by ICP-AES. The cross-checking of the data, specifically by comparing those of Ca, Cr, Fe and Zn, obtained by the two techniques was found to be in good agreement. Special attention from analytical and nutritional point of view has been devoted to the toxic metals. The measurements have been carried out employing the reference algal material prepared by International Atomic Energy Agency (IAEA).     

Comparison of instrumental neutron activation analysis of geological materials with other multielement techniques

Results obtained for 12 elements in approximately 1600 rocks by instrumental neutron activation analysis (INAA) are compared with those obtained by ICP emission spectrometry (ICP-ES), XRF, and atomic-absorption spectrometry (AAS). Sample duplicates and two controls are used to evaluate the precison of the methods investigated. Application of a method (Maximum Likelihood Functional Relationship) to determine and quantify rotational and translational bias is demonstrated. The elements Na, Fe, Ba, Co, Cr, La, Ni and Rb can be determined in rocks by INAA with sufficient sensitivity and precision, whereas the determination of Ag, Yb, Zn and Zr suffers from inadequate sensitivity. Good agreement is seen in the results for Na (by INAA, ICP-ES and XRF) and AG (INAA and AAS). A significant positive bias (13% or less) is evident in the comparison of results by INAA and ICP-ES or XRF for Cr, Ba, Ni and Fe over a wide range of concentration. A similar trend, though less significant, is observed for the elements Yb, Rb, La and Co; the upper limit of concentration for satisfactory determination is within a decade of the highest detection limit for these elements. Rotational and translational bias is evident for Zn in the comparison of data obtained by INAA and ICP-ES, the results by INAA being appreciably lower above about 400 ppm Zn.     

Trace elements in sickle cell disease

Instrumental Neutron Activation Analysis (INAA) and Proton-Induced X-ray Emission (PIXE) analysis (employed as a complementary technique) have been used to determine the concentration of 11 elements in blood samples and its components erythrocytes and plasma obtained, from three groups of subjects in Nigeria viz: sickle cell anaemia (SCA) subjects, subjects with sickle cell trait and normal control subjects. The results suggest that SCA subjects have significantly higher concentrations of Na, Cl, Ca and Cu in their whole blood and erythrocytes and a higher concentration, of Cl and Cu in their plasma relative to the control subjects. Furthermore, a significantly lower concentration of K, Fe, Zn, Se, Br and Rb were found in the whole blood and erythrocytes of the SCA subjects as compared to the controls while the concentration of K and Fe in the plasma of the SCA subjects were however, found to be significantly higher than that of the control group. The study also shows that there was no significant differences between the concentration of these 11 elements in the group with sickle cell trait and the normal control group.     

Elemental analysis of dust trapped in air conditioner filters for the assessment of Lahore city’s air quality

A study was undertaken to assess the air quality of Lahore by the elemental analysis of air conditioner (AC) filter dust samples collected from 15 different commercial sites. Samples were prepared using the Leeds Public Analyst Method and were analyzed using instrumental neutron activation analysis (INAA) for up to 31 elements. The elements Al, As, Ba, Ce, Co, Cr, Cs, Fe, Hf, K, La, Lu, Mn, Na, Nd, Rb, Sc, Sm, Sn, Ta, Th, Yb and Zn were detected in all 15 samples whereas the remaining elements have been detected in fewer samples; i.e. Mg, Sb and Tb were detected in 14 samples, Br and V in ten samples, U in nine samples and Ca and Ti in eight samples only. Al, Ca, Fe, K, Mg and Na were determined in all samples at percentage levels. The concentrations of most elements were found to lie around the mean values for the 15 samples studied and were not orders of magnitude different. However the concentrations of Ca, Mg, Sn and Zn were found to be more variable and were found to be dependant on activities such as construction, fruit and vegetable handling, tin plating and transport, respectively.     

Factorial discriminant analysis of occupational exposure in metallurgy using INAA of hair samples

A multivariate statistical technique-factoral discriminant analysis (FDA)-has been used to evaluate the occupational esposure of a group of metallurgy workers using the elemental composition of their scalp hair, as compared with a control group. The initial variables were the concentrations of ten minor and trace elements Al, Co, Cu, Fe, Mg, Mn, Sb, Se, V, and Zn determined in hair samples by using instrumental neutron activation analysis (INAA). A good discrimination between exposed and control subjects has been achieved after the normalization of the data (by log-transforming the initial variables) and considering sex-matched groups. To measure the accuracy of the classification and the predictive ability a cross-validation procedure was used.     

Differences in trace element concentrations between Alzheimer and “normal” human brain tissue using instrumental neutron activation analysis (INAA)

Brain samples obtained from the Netherlands Brain Bank were taken fromthe superior frontal gyrus, superior parietal gyrus and medial temporal gyrusof 'normal' and Alzheimer's disease subjects in order todetermine elemental concentrations and compare elemental composition. Brainsamples from the cortex were taken from 18 subjects, eight 'normals'(6 males and 2 females) and eleven with Alzheimer's disease, (1 maleand 10 females) and the following elemental concentrations, Na, K, Fe, Zn,Se, Br, Rb, Ag, Cs, Ba, and Eu were determined by instrumental neutron activationanalysis (INAA). The element which showed the greatest difference was Br,which was found to be significantly elevated in the cortex of Alzheimer'sdisease brains as compared to the 'normals' at significance (p<0.001).     

Assessing soil pollution from a municipal waste dump in Islamabad, Pakistan: a study by INAA and AAS

INAA and AAS techniques have been employed to determine 40 elements in soil of a municipal waste dump in sector H-11, Islamabad. Background soil was also analyzed to study the extent of contamination of the dump site soil. Most of the major elements in these soils represented the geochemical composition of the soil in this area. The enrichment factors for quantified elements identified high Sb and Mg contents that could be attributed to the presence of PET and food materials in the waste. Geo-accumulation Index (I _geo), Pollution Index (PI) and the Integrated Pollution Index (IPI) have also been calculated for all elements. The values for these indices show that municipal waste has distorted the soil ambiance and the soil of waste dump site is slightly to moderately polluted as compared to the background soil. The dump soil was found to be moderately polluted by the elements Ba, Br, Ga, Rb, Zn, Ni and Pb. Significantly high Cu, Mg and Sb contamination was observed for the waste soil that is likely to pose an environmental issue if current waste disposal procedures are continuously employed.     

Characterisation of biological RMs for potential use in human brain analysis

Summary In order to investigate the precision and accuracy obtainable with instrumental neutron activation analysis (INAA) and inductively coupled plasma atomic emission spectrometry (ICP-AES), five different biological reference materials were analyzed. Four of them originated from the US National Institute of Standards and Technology (NIST) (Bovine Liver (SRM 1577; 1577/a; 185/A), Orchard Leaves (SRM 1571)) and one of them came from the International Atomic Energy Agency (IAEA) (Animal muscle (H-4)). The decomposition efficiency of microwave acid digestion was examined in comparison to digestions carried out in a Parrbomb. Five to ten subsamples of each material were subjected to the two digestion procedures and the solutions were analyzed by ICP-AES. The dried samples were analyzed by INAA. Up to 16 elements (Al, B, Ca, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, S, Zn) were determined in various materials.     

Determination of Ca, Mg, Fe, Cu, and Zn in blood fractions and whole blood of humans by ICP-OES

Inductively coupled plasma – optical emission spectrometry (ICP-OES) was applied to the determination of the elements Ca, Mg, Fe, Cu, and Zn in blood plasma, erythrocytes, lymphocytes, and whole blood to obtain reliable data on their distribution in blood fractions. The samples were carefully collected to avoid contamination. Two different nebulizers (Babington and Meinhard) were tested and optimized for this analytical problem. Line selections for all elements of interest were performed (LODs were 0.8 μg/L for Ca, 1.7 μg/L for Cu, 3.0 μg/L for Fe, 1.1 μg/L for Mg, and 4.2 μg/L for Zn). Recoveries were determined as approx. 100%, and standard reference material was analyzed to obtain reliable data on element distribution. The optimized method was applied to the determination of Ca, Mg, Fe, Cu, and Zn in the course of a clinical study on blood and blood fractions of two groups of humans of differing health. The concentrations measured in blood fractions were verified by balancing with the values found in whole blood.     

Elemental composition of hair and bone density measurements at diagnosis in paediatric inflammatory bowel disease

A dual energy X-ray absorptiometry (DEXA) system has been employed to measure the bone mineral density (BMD) of children with inflammatory bowel disease (IBD); Crohn's disease (CD) and ulcerative colitis (UC) at diagnosis. The results of the study indicate that even at diagnosis, a significantly reduced bone mineral density measured in terms of the Z-score, was observed in the lumbar spine and whole body of the IBD subjects. This was found to be more pronounced in Crohn's disease than in the ulcerative colitis subjects. Furthermore, the study was extended to investigate the elemental composition of hair of IBD subjects and age matched controls using instrumental neutron activation analysis (INAA). The concentrations of 11 elements; Al, Br, Ca, Cl, Cu, I, K, Mg, Mn, Na and V were determined. A significantly reduced concentration of Ca, Cu, K, Mn, and V were observed in the hair of IBD subjects as compared to the control group. This revised version was published online in August 2006 with corrections to the Cover Date.     

Evaluation of concentrations of major and trace elements in human lung using INAA and PIXE

The elemental concentrations of Br, Ca, Ce, Cl, Co, Cr, Cs, F, Fe, Hf, K, Mg, Mn, Na, O, Rb, Sb, Sc, Se, V and Zn in 15 human lung autopsy samples, taken from subjects aged more than fifty years old, were determined by instrumental neutron activation analysis (INAA) using reactor neutrons in conjunction with a high resolution detection system. Two modes of irradiation and counting were applied; namely cyclic neutron activation analysis (CNAA) and conventional neutron activation analysis. Proton induced X-ray emission (PIXE) analysis, using a proton beam emerging from a 2 MV Van de Graff accelerator, was additionally employed and Ge, Ni, P and Ti were also identified in the lung tissue. Detection of the X-ray spectra was performed using a high resolution Si(Li) semiconductor. The relevance of these results, including a comparison between the concentrations of elements measured in a pig's lung using CNAA and those found in the human lung is discussed.     

Simultaneous determination of Fe, P, Ca, Mg, Zn, and Cu in whole blood by two-jet plasma atomic emission spectrometry

Availability of many biological samples in ample quantity for biomedical investigations sometimes is very restricted. Therefore, there is the need for the simple techniques allowing the analysis of small amount samples. In the present work the two-jet plasma atomic emission spectrometry techniques for the determination of Fe, P, Ca, Mg, Zn, and Cu in whole blood are proposed. The first technique is developed for direct analysis of freeze-dried blood. The sample preparation consisted in a dilution of blood powder (particle size 20 μm or less) with a spectroscopic buffer (graphite powder containing 15 wt.% NaCl). For the analysis of liquid whole blood, previous carbonization (not ashing) of blood evaporated on graphite powder was applied. Calibration samples based on graphite powder containing 15 wt.% NaCl were used. The validation of the techniques was confirmed by the use of different sample preparation procedures (wet acid digestion and dry carbonization), the analysis of IAEA A-13 reference material (freeze-dried bovine whole blood), and the comparison of the results obtained by the proposed technique with the results of the stripping voltammetry technique. Just 20-50 μL of whole blood is quite enough for all determinations. The proposed techniques were successfully applied for the simultaneous determination of Fe, P, Ca, Mg, Zn, and Cu in whole blood of living experimental rats and mice and human blood.