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.     

Direct determination of selenium and other trace elements in serum samples by ICP-MS.

Selenium belongs to a group of trace elements of special interest in biological samples for clinical diagnosis. Selenium has antioxidizing functions and is essential for providing the organism with triiodothyronine produced from thyroxine. Among several analytical techniques used to determine the Se concentration in serum, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) has been used in the past because of its high sensitivity. Interference problems originating from different ions on the major Se isotopes have been described to be a limiting factor for the direct determination of Se in these matrices. Standard addition calibration or isotope dilution is often required to overcome carbon-enhanced ionisation effects in biological sample matrices. In most cases, the typical serum sample volume which is available for the analysis is limited to 0.5 ml or less, making multiple sample preparation for standard addition calibration impractical. Isotope dilution requires enriched isotopes and substantial sample preparation. Furthermore, the approximate Se concentration in every sample has to be known to adjust the appropriate amount of spike to each sample. Matrix matching with methanol has been described to overcome ionisation effects but we found limiting factors of this application when other trace elements are also determined within one sample run. This paper describes an effective sample preparation method which allows the direct determination of Se in serum without limiting the analytical capabilities for the additional determination of Al, Cu, Ni, Co, Cd, Mn and Zn in a single sample run by ICP-MS. Optimization procedures are presented and results of the analysis of reference samples are discussed, with a comparison of more than 150 serum data with those obtained by the GF-AAS method.     

Zn concentration in thyroid tissue and whole blood of women with different diseases of thyroid

The Zn concentration in thyroid tissue and whole blood of patients with Graves’ disease, thyroid cancer, and nodular goiter disease was determined using the total-reflection X-ray fluorescence method. The dependence of obtained concentrations on the clinical stage of the examined disease, histopathological grading, and kind of analyzed material (thyroid tissue and blood) was studied. The determined concentration of Zn was the lowest in the thyroid tissue of patients with thyroid cancer (23.1 μg/g) and it was the highest in the case of Graves’ disease (41.7 μg/g), whereas in the blood samples, the reverse results were found (7.1 μg/g and 4.8 μg/g, respectively). The physical basis of the method used, the experimental setup, and the procedure of sample preparation are described.     

Reference values for trace and ultratrace elements in human serum determined by double-focusing ICP-MS

Reference values for trace and ultratrace elements concentrations in healthy human serum, measured by double-focusing inductively coupled plasma-mass spectrometry (ICP-MS), are presented. Blood donors from Asturias (Spain) were selected as the reference population (n=59). Blood samples were collected, after donation, taking the necessary precautions to avoid contamination. All subjects analyzed had normal renal function and nutritional status, as shown from their creatinine and albumin levels. A total number of 14 elements (Al, Ca, Cr, Mn, Fe, Co, Cu, Zn, Rb, Sr, Mo, Cd, Pb, and U) were monitored almost simultaneously. Serum samples were diluted 1+4 with ultrapure water and matrix interferences were corrected using Sc, Ga, Y, and Tl as internal standards. Fe, Cu, and Zn were also determined by isotope dilution analysis (IDA). Reference trace element concentrations intervals observed containing 95% of the reference distribution after excluding outliers are presented. Fourteen serum samples from hemodialysis patients were also analyzed for comparison. High levels of Al, Cr, Sr, Mo, Mn, Pb, U, Co, and Cu and low levels of Fe, Zn, and Rb were found in the serum samples from hemodialysis patients compared to the corresponding reference values observed in this work.     

Essential trace elements in humans

Serum arsenic concentrations of persons suffering from renal failure and undergoing hemodialysis treatment (n=85) and of healthy controls (n=25) were determined by hydride-generation AAS technique after microwave digestion. The results were evaluated by comparing the values of both groups, considering physiological factors and individual data, as well as comorbid conditions of the hemodialysis (HD) patients. Serum arsenic levels were diminished in the patient group compared with controls (mean values 8.5±1.8 ng/mL vs 10.6±1.3 ng/mL). Furthermore, additional diseases within the hemodialysis group, particularly injuries of the central nervous system (CNS), vascular diseases, and cancer, were correlated to occasionally markedly decreased serum arsenic concentrations. It was concluded that arsenic homeostasis is disturbed by HD treatment and certain additional diseases. Desirable arsenic concentrations in the body seem to be reasonable. This consideration results in the conclusion that arsenic could play an essential role in human health. Thus, reference arsenic concentrations in different human tissues and body fluids should be established in order to recognize not only arsenic intoxication, but also arsenic deficiency. Perhaps arsenic deficiency contributes to the increased death risk of HD patients, and therefore, arsenic supplementations for patients with extremely low serum arsenic concentrations should be taken into account.     

Heroin dependence effects on some major and trace elements

In this study, 56 (14 control and 42 addicts) adult human subjects of both sexes of different periods of heroin dependence were subjected to the measurement of whole blood, serum, and red blood cell levels of some trace elements (zinc, managanese, iron copper, and bromine), as well as some major elements (phosphorus, sulfur, calcium, potassium, and chlorine). This was done by the energy-dispersive X-ray fluorescence (EDXRF) technique, in which copper and bromine showed a significant rise in whole blood (male) (22 and 32%, respectively), while zinc, iron, manganese, calcium, sulfur phosphorus, potassium, and chlorine showed a significant drop (49, 8, 25, 34, 21, 51, 61 and 72%, respectively) in proportion to the period of heroin intake (6 yr) and in comparison with the control group. No significant sexual variation has been reported.     

Essential trace elements in milk and blood serum of lactating donkeys as affected by lactation stage and dietary supplementation with trace elements

The aim of this trial was to study the concentration of zinc (Zn), iron (Fe), copper (Cu), manganese (Mn), selenium (Se), cobalt (Co) and iodine (I) in milk and blood serum of lactating donkeys, taking into account the effects of lactation stage and dietary supplementation with trace elements. During a 3-month period, 16 clinically healthy lactating donkeys (Martina-Franca-derived population), randomly divided into two homogeneous groups (control (CTL) and trace elements (TE)), were used to provide milk and blood samples at 2-week intervals. Donkeys in both groups had continuous access to meadow hay and were fed 2.5 kg of mixed feed daily, divided into two meals. The mixed feed for the TE group had the same ingredients as the CTL, but was supplemented with a commercial premix providing 163 mg Zn, 185 mg Fe, 36 mg Cu, 216 mg Mn, 0.67 mg Se, 2.78 mg Co and 3.20 mg I/kg mixed feed. The concentrations of Zn, Fe, Cu, Mn, Se, Co and I were measured in feeds, milk and blood serum by inductively coupled plasma-MS. Data were processed by ANOVA for repeated measures. The milk concentrations of all the investigated elements were not significantly affected by the dietary supplementation with TE. Serum concentrations of Zn, Fe, Cu Mn and Se were not affected by dietary treatment, but TE-supplemented donkeys showed significantly higher concentrations of serum Co (1.34 v. 0.69 μg/l) and I (24.42 v. 21.43 μg/l) than unsupplemented donkeys. The effect of lactation stage was significant for all the investigated elements in milk and blood serum, except for serum manganese. A clear negative trend during lactation was observed for milk Cu and Se concentrations (−38%), whereas that of Mn tended to increase. The serum Cu concentration was generally constant and that of Co tended to increase. If compared with data reported in the literature for human milk, donkey milk showed similarities for Zn, Mn, Co and I. Furthermore, this study indicated that, in the current experimental conditions, the mineral profile of donkey milk was not dependent on dietary TE supply.     

Concentrations of trace elements in osteoarthritic knee-joint effusions

Concentrations of the 18 elements, barium (Ba), beryllium (Be), bismuth (Bi), calcium (Ca), cadmium (Cd), cesium (Cs), copper (Cu), lanthanum (La), lithium (Li), magnesium (Mg), molybdenum (Mo), lead (Pb), rubidium (Rb), antimony (Sb), tin (Sn), strontium (Sr), thallium (Tl), and zinc (Zn), were determined in the synovial fluids of osteoarthritic knee joints and in the corresponding sera of 16 patients by inductively coupled plasma-mass spectrometry. Knee-joint effusions have lower elemental concentrations than their corresponding sera. For the essential elements Ca, Cu, Mg, and Zn and for the nonessential and toxic elements Ba, Be, Bi, La, and Sb, this difference was highly significant. Strong positive correlations between concentrations in effusions and sera for the essential elements Cu and Mg and for the nonessential elements Cs, Li, Rb, and Sr could be established. The grade of localized hyperperfusion of the knee region in the blood pool phase of ^99mTc HDP bone scan indicating inflammation did not correlate with any elemental concentration determined. Deceased.     

Studies on some trace and minor elements in blood

Blood is one of the widely used specimens for biological trace element research because of its biological significance and ease of sampling. We have conducted a study of the blood of the Kalpakkam township population for trace and minor elements. For this purpose, analytical methods have been developed and standardized in our laboratory for the elemental analysis of blood plasma and red cells. Inductively coupled plasma-mass spectrometry (ICP-MS), a relatively new technique, has been applied for the analysis of trace elements. Details regarding spectral interference and matrix interference encountered in the analysis of blood and the methods of correcting them have been discussed. Flame atomic absorption spectrometry (AAS)/atomic emission spectrometry (AES) has been applied for the determination of minor elements. Precision and accuracy of these methods have also been discussed.     

Differentiation of serum levels of trace elements in normal and malignant breast patients

The objective of this study was to determine which elements in serum best differentiated breast cancer in a case-control study. Concentrations of 13 elements in serum of 68 breast tumor patients (25 malignant and 43 benign) and 26 healthy controls were measured. Logistic regression with different variable-selection procedures was used to determine a possible configuration of elements. Sensitivity and specificity of the model were calculated to obtain the optimal cutoff point for discriminating malignant breast cancers vs other individuals (including benign breast disease and normal ones). A combination of Cd, Mn, and Fe was found to have a specificity and sensitivity of 100% using forward-type logistic regression, when the cutoff value of the combination score was 52.71. Using stepwise-type logistic regression, a combination of Cr and Mn had a sensitivity of 100% and a specificity of 97.1% when the combination score of 17.4 was chosen as the cutoff. Similar analysis could be implemented to compare the malignant and control groups. Specificity and sensitivity were 100% for Mn (forward and stepwise type) with a cutoff point of 6.40. For the backward regression, specificity was 84.6% and sensitivity was 100% for Zn, with a cutoff point of 869.1. In conclusion, there was a significant difference in concentrations of all 13 elements in serum between breast cancer patients and controls. A combination among Cd, Mn, Fe, Cr, and Zn might be important to determine a differentiating reference for breast cancers if a long-term followed-up study is to be conducted.     

Minor and trace elemental determination in the Indian herbal and other medicinal preparations

Medicinal plants described in the Indian “Ayurvedic” literature viz. Tulsi (Ocimum sanctum), Gulvel (Tinospora cardifolia), bitter Neem (Azadirachta indica), Kanher (Nerium Åndicum), Vekhand (Acorus calamus), and Peacock's feather (ash) were analyzed for minor and trace elements by instrumental neutron activation analysis. The samples and the standards from the National Institute of Standards and Technology, USA and IAEA, Vienna were irradiated for 5 min, 1 h, 5 h, and 10 h with thermal neutrons at a flux of 10¹²–10¹³ n cm^?2s^?1 in APSARA and CIRUS reactor at BARC Bombay. High resolution γ ray spectrometry was performed using a 45 cm³ HPGe detector and a 4096 MCA system. Concentrations of 13 elements were determined. Zinc, manganese, and sodium were significantly higher in Tulsi leaves while zinc is higher in Neem leaves. Peacock's feathers were found to be rich in manganese, iron, copper, and zinc. A high concentration of mercury was also found in the peacock's feather ash. The therapeutic significance in restoring ionic balance is discussed.     

Serum concentrations of macro and trace elements in heroin addicts of the Canary islands

Na, K, Ca, Mg, P, Fe, Cu, Zn and Se concentrations were determined in the serum of 106 heroin addicts and were compared with the concentrations obtained in a control group formed of 186 apparently healthy individuals. Heroin addicts displayed K and Se mean concentrations lower (p < 0.05), and Na, Mg, P mean concentrations and a Cu/Zn ratio higher (p < 0.05) than those mean values observed in the control group. The Mg and P concentrations in the serum of heroin addicts tended to normalize when age increased. The heroin addicts included in the methadone maintenance treatment program had higher serum mean concentrations of K and Mg than the heroin addicts in the detoxification process. The Na, K and Mg concentrations displayed highly significant correlations, with a different behavior for the heroin addicts group and the control group. When applying factor analysis and representing the scores of the first and second factors, the heroin addicts tended to differentiation from the control group. However, methadone substitution treatment was not able to normalize these concentrations.     

Role of trace elements in primary arterial hypertension

A negative relationship between water hardness and cardiovascular mortality rate was demonstrated and became a source of interest regarding minerals and trace metals in the pathogenesis of atherosclerosis, cardiovascular diseases, and arterial hypertension. Higher incidences of sudden death, cerebrovascular diseases, arterial hypertension, and coronary heart disease have been reported in soft water areas. A major research effort has been devoted to the problem in an attempt to find a protective factor in hard water or a detrimental factor or element in soft water. The roles of calcium, magnesium, cobalt, lithium, vanadium, silicon, manganese, and copper have been considered potentially beneficial, whereas those of cadmium, lead, silver, zinc, and antimony have been considered potentially harmful. Cobalt and zinc have been attributed both roles. In the present article, the role of trace quantities of several elements in mineral water in the etiopathogenesis of primary arterial hypertension is reviewed. An erratum to this article is available at .     

氮磷肥对茶树锌硒等中微量元素吸收与分配的影响

锌（Zn）和硒（Se）及其他中微量元素（铝Al,钙Ca,铁Fe,铜Cu,锰Mn）是茶叶品质的重要指标,但茶树吸收Zn、Se能力及氮（N）磷（P）肥影响中微量元素吸收与分配的过程尚不清楚。以红壤丘陵区福鼎大白茶树为研究对象,开展Zn+Se、Zn+Se+N、Zn+Se+P、Zn+Se+N+P和对照共5种处理3次重复随机化区组试验,处理第3年春季分茶叶、成熟叶、吸收根、运输根和储藏根采集植物样品,测定其元素含量。结果表明,茶树地上和地下器官Zn和Se及其他中微量元素对N、P、Zn、Se添加的响应具分异性。与对照相比,茶树地上和地下器官Zn和Se含量均显著增加,与Zn+Se相比,施N和/或P肥仅显著提高茶叶和成熟叶Se含量（P<0.05）;与对照相比,施肥处理均显著提高吸收根和运输根Al、Fe含量以及储藏根Cu含量;运输根Mn含量表现为Zn+Se+N、Zn+Se+P、Zn+Se+N+P显著高于对照,储藏根Mn含量为Zn+Se+N+P显著高于其他处理;茶树各器官Ca含量对施肥处理无显著响应。此外,茶叶和成熟叶的Zn含量与吸收根显著正相关,而Se含量则与储藏根显著正相关。茶树具有吸收和积累Zn和Se的能力,而施N、P肥有助于提高茶叶Se含量,研究结果为红壤丘陵区培育高品质锌硒茶及营建生态高值茶园提供了依据。     

INAA of trace elements in colorectal cancer patients

The concentration of trace elements in samples of both colorectal cancer tumors and normal tissues of a Mexican population were irradiated for 30 s and 4 h and their elemental content were measured by instrumental neutron activation analysis. Ca, Cu, Co, I, Mg, Se, Fe, Zn, Hg, Ba, and Cr were analyzed. Alterations in Co, Fe, I, and Ba were found in tumors with respect to normal tissues.     

Major and trace elements in whole blood of phlebotomized patients with porphyria cutanea tarda

Porphyria cutanea tarda (PCT) is a disorder of hem biosynthesis resulting from a decreased activity of the uroporphyrinogen decarboxylase enzyme. Hem precursors are accumulated in the blood, liver and skin. Inherited and acquired factors also contribute to the pathogenesis of PCT. Hem precursors and porphyrins are excreted with urine and faeces. Whole blood of 8 PCT patients and 6 volunteers of Caucasian origin were analysed. In addition to routine laboratory measurements, 19 elements (Al, B, Ba, Ca, Cd, Co, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, P, S, V, Zn) were determined by means of inductively coupled plasma optical emission spectrometry (ICP-OES). Mg, P and S concentrations in whole blood were decreased significantly (p<0.05), whereas Ba was increased in PCT patients compared to controls. Metabolic alterations are reflected in the correlation of parameters. Positive correlations were found between the element pairs of Zn-Al, Zn-Mg, Zn-Mn, B-S, Fe-Mg, K-P, Mg-Mn for PCT patients, whereas in the control group Al-Mn, Ca-Cu, Ca-Na, Cu-Mg, Fe-K, Mg-Na, Zn-P showed positive correlations.     

Calcium,magnesium, and other elements in the red blood cells and hair of normals and patients with premenstrual syndrome

This study compared the levels of 18 red cell elements and 22 hair elements in 46 patients (median age: 36.2 yr) diagnosed with PMS (premenstrual syndrome) to 50 normals (median age: 37.7 yr). Significantly lower amounts of calcium, chromium, copper, and manganese were found in the blood of patients with PMS. The ratios of Mg/Ca and K/Na and toxic metals such as lead, arsenic, and germanium were significantly elevated in the PMS patients. In hair, mercury and the Zn/Cu ratio were significantly greater in the PMS patients than the controls, but iron, potassium, and the Mg/Ca ratio were lower. The highly significant Mg/Ca ratio in blood cells may be indicative of a more complex relationship between PMS and magnesium and calcium than either element alone. The significantly lower blood cell calcium level found in these studies may provide additional evidence that PMS may be related to a calcium-deficiency state or a metabolic defect involving calcium.     

Levels of some trace elements and rheumatoid factor in sheep with brucellosis

Brucellosis is a zoonotic disease that is encountered in sheep rather frequently. In this study, 100 sheep diagnosed with brucellosis that had aborts and 40 healthy sheep were used as materials. Analyses for Cu, Zn, Fe, Cr, Ca, Mg, and K were performed with the atomic absorption spectrophotometric method on blood collected from vena jugularis of all the sheep and rheumatoid factor levels were determined by the nephelometry method. Although it was found that Cu, Ca, and rheumatoid factor values in the sera of the sheep with brucellosis were high when compared to the control group (p<0.001, p<0.05, p<0.001, respectively), their serum Zn values were low (p<0.05). No significant changes in serum Cr, Fe, K, and Mg levels were found.     

Level of trace elements in serum and toenail samples of patients with onychocryptosis (ingrown toenail) and onychomycosis

BackgroundOnychocryptosis (ingrown toenail) and onychomycosis are common pathologies of the toenail and affecting many people. Since levels of trace elements have been shown to vary in certain diseases, in the presented work, chromium (Cr), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), selenium (Se), and zinc (Zn) levels of toenail and serum samples of healthy individuals and patients with onychocryptosis and onychomycosis were compared.MethodsSerum and toenail samples of 88 voluntary subjects (healthy n = 24; onychomycosis n = 24; onychocryptosis n = 40) aged between 19–80 years were collected. Levels of trace elements in the samples were analyzed by using an inductively coupled plasma-optical emission spectrophotometer (ICP-OES Thermo iCAP - 6000). The differences in medians between the groups for elements were evaluated with Kruskal -Wallis H test with post hoc for pairwise comparisons in SPSS 18.ResultsMg (p < 0.001) and Mn (p = 0.002) levels were significantly increased whereas Zn (p = 0.011) level was decreased in toenails of patients with onychomycosis compared to healthy subjects. Although Mg and Mn levels were higher in female subjects with onychomycosis (p = 0.001; p = 0.019), Mn was only increased in male subjects (p = 0.015). Mg was the only trace element found to be independent of sex, age, and smoking status in patients with onychomycosis. However, no significant difference has been found in serum trace element levels neither between any groups nor toenail trace element levels of patients with onychocryptosis and healthy subjects.ConclusionAs a response of the human body to pathogens like fungi in toenails, Mg, Mn and Zn levels vary. Especially the role of Mg ions in onychomycosis needs to be investigated more specifically.     

Direct determination of selenium in rat blood plasma by Zeeman atomic absorption spectrometry

The method was developed to be applied for direct determination of selenium in rat plasma by graphite-furnace atomic absorption spectrometry with Zeeman background correction. Blood was obtained from CD rats of both sexes 2h after dosing in weeks 7 and 13 in order to acquire data on the levels of selenium in these animals during 13-week gavage administration of l-seleno-methylselenocysteine (SeMC), a new candidate chemopreventive agent under development. Application of the commonly used method of standard addition was found to be unsuitable to calculate the selenium content in rat plasma (within-run and between-run accuracy and precision parameters were less than 85%). Therefore, a new analytical method was developed. In this method, samples of rat plasma (50 microL) were diluted 10-fold with a reducing agent containing l-ascorbic acid, a modifier solution containing palladium chloride and Triton X-100. Samples were atomized in pyrolytically coated graphite tubes and peak height signals were measured. Selenium concentrations were determined by linear least squares regression analysis based on the standard curve generated in pooled rat blank plasma. Since selenium is normally present in plasma, a three-step approach was used to calculate selenium plasma levels. Initially selenium levels were determined based on the standard curve with selenium-spiked pool plasma. In the second step, background selenium levels in the pooled plasma were determined based on the same standard curve. In the third step, background level was added to the previously derived number. The relative errors were in the range from -4.6 to 11.4% (intra-day assay) and from -0.4 to 8.8% (inter-day assay) which proved good accuracy. The relative standard deviations were in the range from 1.88 to 4.70% (intra-day precision) and from 3.28 to 5.38% (inter-day precision). In rat plasma, the following dose-dependent selenium levels (mean+/-S.D.) in males and females, respectively, were observed at 13 weeks: 655.5+/-48.8 and 595.8+/-43.9 ng/mL (control group), 927.9+/-85.3 and 859.3+/-164.3 ng/mL (0.4 mg/kg per day dose group), 1238.9+/-182.4 and 1169.9+/-112.6 ng/mL (0.8 mg/kg per day dose group), and 1476.5+/-138.1 and 1320.1+/-228.6 ng/mL (2.0mg/kg per day dose group). No significant sex differences in selenium plasma levels were seen in the SeMC-treated groups. No significant differences in selenium plasma levels were seen between mean plasma levels at 7 and 13 weeks. The described method is simple, rapid, accurate, precise and can be easily applied in other laboratories for a large number of samples.