Inductively Coupled Plasma-Mass Spectrometric Determination of Iodine in Food Using Tetramethyl Ammonium Hydroxide Extraction-Results from a U.S. Food and Drug Administration Level 3 Interlaboratory Study.

Background: The performance of U.S. Food and Drug Administration (FDA) Elemental Analysis Manual (EAM) 4.13 method (Inductively Coupled Plasma-Mass Spectrometric Determination of Iodine in Food Using Tetramethyl Ammonium Hydroxide Extraction) was tested in an interlaboratory study. Objective: The aim of the study is to demonstrate that the FDA EAM method 4.13 is applicable for the analysis of food and multivitamins. Methods: Six collaborators participated in the study using four different models of inductively coupled plasma-mass spectrometry instruments. The method evaluation included determination of the limits of detection and quantification, analysis of National Institute of Standards and Technology standard reference materials (SRMs), unknown samples, blinded SRMs, and fortified analytical portions by all six collaborators. The samples were chosen to represent all sectors of the AOAC food triangle and additionally included pet food and multivitamin tablets. Results: The repeatability and reproducibility ranges were 1.8-11.4% and 3.6-13.7%, respectively; the calculated HorRat values were in the 0.17-1.18 range; and 174 of 175 SRM analyses had z-scores <2 and fortified analytical portion samples with recoveries of 102-105%, indicating acceptable method performance. Conclusions: The study supports a Level Three Multilaboratory Validation according to FDA Food and Veterinary Program Guidelines performed by six collaborators using six certified reference materials and nine unknown samples. Highlights: The method is applicable for quantification of the total extractable iodine in food and multivitamin dietary supplements.

Semen Uranium Concentrations in Depleted Uranium Exposed Gulf War Veterans: Correlations with Other Body Fluid Matrices.

Environmental metal exposure, as well as dietary metals, may adversely affect semen quality even as others play an essential role in normal spermatogenesis and fertility. Measures of seminal fluid metals have therefore been of high interest in the last several decades but have shown inconsistent results in correlations with some semen quality parameters. As well, environmental metal measures across various body fluid matrices have not been consistently correlated contrary to what one might hypothesize based on a systemic body burden of metal. This may be due to the body fluid matrices assessed and to other differences in laboratory methods and sample preparation. Measures of uranium, a potentially toxic metal in humans, have not previously been reported in the semen of environmentally metal-exposed populations. We report here uranium seminal fluid results and the high correlation of uranium concentrations across several body fluid matrices in a cohort of military veterans exposed to depleted uranium in combat events during the Iraqi Gulf War. These results inform the risk communication conversation for exposed populations and broaden the public health assessments from various exposure scenarios.

Uranium quantification in semen by inductively coupled plasma mass spectrometry.

In this study we report uranium analysis for human semen samples. Uranium quantification was performed by inductively coupled plasma mass spectrometry. No additives, such as chymotrypsin or bovine serum albumin, were used for semen liquefaction, as they showed significant uranium content. For method validation we spiked 2g aliquots of pooled control semen at three different levels of uranium: low at 5 pg/g, medium at 50 pg/g, and high at 1000 pg/g. The detection limit was determined to be 0.8 pg/g uranium in human semen. The data reproduced within 1.4-7% RSD and spike recoveries were 97-100%. The uranium level of the unspiked, pooled control semen was 2.9 pg/g of semen (n=10). In addition six semen samples from a cohort of Veterans exposed to depleted uranium (DU) in the 1991 Gulf War were analyzed with no knowledge of their exposure history. Uranium levels in the Veterans' semen samples ranged from undetectable (<0.8 pg/g) to 3350 pg/g. This wide concentration range for uranium in semen is consistent with known differences in current DU body burdens in these individuals, some of whom have retained embedded DU fragments.

Spatial mapping of mineralization with manganese-enhanced magnetic resonance imaging.

Paramagnetic manganese can be employed as a calcium surrogate to sensitize the magnetic resonance imaging (MRI) technique to the processing of calcium during the bone formation process. At low doses, after just 48h of exposure, osteoblasts take up sufficient quantities of manganese to cause marked reductions in the water proton T1 values compared with untreated cells. After just 24h of exposure, 25µM MnCl(2) had no significant effect on cell viability. However, for mineralization studies 100µM MnCl(2) was used to avoid issues of manganese depletion in calvarial organ cultures and a post-treatment delay of 48h was implemented to ensure that manganese ions taken up by osteoblasts is deposited as mineral. All specimens were identified by their days in vitro (DIV). Using inductively coupled plasma optical emission spectroscopy (ICP-OES), we confirmed that Mn-treated calvariae continued to deposit mineral in culture and that the mineral composition was similar to that of age-matched controls. Notably there was a significant decrease in the manganese content of DIV18 compared with DIV11 specimens, possibly relating to less manganese sequestration as a result of mineral maturation. More importantly, quantitative T1 maps of Mn-treated calvariae showed localized reductions in T1 values over the calvarial surface, indicative of local variations in the surface manganese content. This result was verified with laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). We also found that ΔR1 values, calculated by subtracting the relaxation rate of Mn-treated specimens from the relaxation rate of age-matched controls, were proportional to the surface manganese content and thus mineralizing activity. From this analysis, we established that mineralization of DIV4 and DIV11 specimens occurred in all tissue zones, but was reduced for DIV18 specimens because of mineral maturation with less manganese sequestration. In DIV25 specimens, active mineralization was observed for the expanding superficial surface and ΔR1 values were increased due to the mineralization of small, previously unmineralized areas. Our findings support the use of manganese-enhanced MRI (MEMRI) to study well-orchestrated mineralizing events that occur during embryonic development. In conclusion, MEMRI is more sensitive to the study of mineralization than traditional imaging approaches.

Prostate cancer outcome and tissue levels of metal ions.

BACKGROUND: There are several studies examining prostate cancer and exposure to cadmium, iron, selenium, and zinc. Less data are available on the possible influence of these metal ions on prostate cancer outcome. This study measured levels of these ions in prostatectomy samples in order to examine possible associations between metal concentrations and disease outcome. METHODS: We obtained formalin fixed paraffin embedded tissue blocks of prostatectomy samples of 40 patients with PSA recurrence, matched 1:1 (for year of surgery, race, age, Gleason grading, and pathology TNM classification) with tissue blocks from 40 patients without recurrence (n = 80). Case-control pairs were compared for the levels of metals in areas adjacent to tumors. Inductively coupled plasma-mass spectrometry (ICP-MS) was used for quantification of Cd, Fe, Zn, and Se. RESULTS: Patients with biochemical (PSA) recurrence of disease had 12% lower median iron (95 µg/g vs. 111 µg/g; P = 0.04) and 21% lower zinc (279 µg/g vs. 346 µg/g; P = 0.04) concentrations in the normal-appearing tissue immediately adjacent to cancer areas. Differences in cadmium (0.489 µg/g vs. 0.439 µg/g; 4% higher) and selenium (1.68 µg/g vs. 1.58 µg/g; 5% higher) levels were not statistically significant in recurrence cases, when compared to non-recurrences (P = 0.40 and 0.21, respectively). CONCLUSIONS: There is an association between low zinc and low iron prostate tissue levels and biochemical recurrence in prostate cancer. Whether these novel findings are a cause or effect of more aggressive tumors, or whether low zinc and iron prostatic levels raise implications for therapy, remains to be investigated.

Enzyme stabilization by glass-derived silicates in glass-exposed aqueous solutions.

OBJECTIVES: To analyze the solutes leaching from glass containers into aqueous solutions, and to show that these solutes have enzyme activity stabilizing effects in very dilute solutions. METHODS: Enzyme assays with acetylcholine esterase were used to analyze serially succussed and diluted (SSD) solutions prepared in glass and plastic containers. Aqueous SSD preparations starting with various solutes, or water alone, were prepared under several conditions, and tested for their solute content and their ability to affect enzyme stability in dilute solution. RESULTS: We confirm that water acts to dissolve constituents from glass vials, and show that the solutes derived from the glass have effects on enzymes in the resultant solutions. Enzyme assays demonstrated that enzyme stability in purified and deionized water was enhanced in SSD solutions that were prepared in glass containers, but not those prepared in plastic. The increased enzyme stability could be mimicked in a dose-dependent manner by the addition of silicates to the purified, deionized water that enzymes were dissolved in. Elemental analyses of SSD water preparations made in glass vials showed that boron, silicon, and sodium were present at micromolar concentrations. CONCLUSIONS: These results show that silicates and other solutes are present at micromolar levels in all glass-exposed solutions, whether pharmaceutical or homeopathic in nature. Even though silicates are known to have biological activity at higher concentrations, the silicate concentrations we measured in homeopathic preparations were too low to account for any purported in vivo efficacy, but could potentially influence in vitro biological assays reporting homeopathic effects.

Analysis of iron, zinc, selenium and cadmium in paraffin-embedded prostate tissue specimens using inductively coupled plasma mass-spectrometry.

Formalin-fixed paraffin-embedded (FFPE) tissue specimens represent a valuable and abundant resource of pathologic material for various biomedical studies. In the present study, we report the application of high-resolution inductively coupled mass-spectrometry (ICP-MS) for quantification of Fe, Zn, Se and Cd in FFPE prostate tissue. These elements have a possible role in the development of prostate diseases: while Zn and Se are needed for a healthy prostate, Cd shows multiple toxic and carcinogenic effects. Excessive accumulation of Fe induces the production of highly reactive hydroxyl radical species, which may play a role in cancer etiopathogenesis. To assess whether the levels of these metals in the FFPE prostate tissue represent their original content, we compared their levels with those in the fresh tissue (on dry weight basis) in samples obtained from 15 patients. We found that in FFPE tissue, the recoveries of Se, Fe, Cd and Zn were progressively decreased, 97+/-11% (r=0.88), 82+/-22% (r=0.86), 59+/-23% (r=0.69) and 24+/-11% (r=0.38), respectively. Thus, the use of correction factors, determined as k=0.16 for Se, k=0.20 for Fe, k=0.27 for Cd and k=0.67 for Zn, is required to estimate the retrospective levels of these elements in the parental non-processed fresh (wet) prostate tissue. The technique used in this study enables the analysis of archival FFPE prostate tissue for the concentrations of Fe, Zn, Se and Cd to study association between the levels of these metals and prostate disease.

Clinical and analytical toxicology of dietary supplements: a case study and a review of the literature.

The use of dietary supplements has grown dramatically in the last decade. A large number of dietary and herbal supplements escape regulatory and quality control; components of these preparations are poisonous and may contain, among other toxins, heavy metals. Uncontrolled use of dietary and herbal supplements by special populations, such as the military, may therefore pose a health risk. Clinical symptoms are not always properly attributed to dietary supplements; patients often do not mention supplement use to their health care provider. Therefore, a health risk estimate is hard to make on either the individual or the population level. The literature on this issue was reviewed and discussed in the light of a representative clinical-chemical case study. This case study was performed on a host of preparations that were used by one single individual in the military. Both essential (chromium, copper, zinc, and iron) and poisonous (arsenic, lead, and nickel) trace elements were determined using inductively coupled plasma combined with optical emission spectrometry (ICP-OES) or with mass spectrometry (ICP-MS). Arsenic and lead were detected at exposure levels associated with health risks. These health risks were detected predominantly in hormone-containing supplements and the herbs and botanicals used for performance enhancement. To the extent that this is a representative sample, there is an underestimation of supplement use and supplement risk in the US military, if not in the general population. Since clinical symptoms may be attributed to other causes and, unless patients are specifically asked, health care providers may not be aware of their patients' use of dietary supplements, a strong support of laboratory diagnostics, such as a toxicological screening of blood or urine, is required. In addition, screening of the preparations themselves may be advised.

Manganese-enhanced magnetic resonance microscopy of mineralization.

Paramagnetic manganese (II) can be employed as a calcium surrogate to sensitize magnetic resonance microscopy (MRM) to the processing of calcium during bone formation. At high doses, osteoblasts can take up sufficient quantities of manganese, resulting in marked changes in water proton T(1), T(2) and magnetization transfer ratio values compared to those for untreated cells. Accordingly, inductively coupled plasma mass spectrometry (ICP-MS) results confirm that the manganese content of treated cell pellets was 10-fold higher than that for untreated cell pellets. To establish that manganese is processed like calcium and deposited as bone, calvaria from the skull of embryonic chicks were grown in culture medium supplemented with 1 mM MnCl(2) and 3 mM CaCl(2). A banding pattern of high and low T(2) values, consistent with mineral deposits with high and low levels of manganese, was observed radiating from the calvarial ridge. The results of ICP-MS studies confirm that manganese-treated calvaria take up increasing amounts of manganese with time in culture. Finally, elemental mapping studies with electron probe microanalysis confirmed local variations in the manganese content of bone newly deposited on the calvarial surface. This is the first reported use of manganese-enhanced MRM to study the process whereby calcium is taken up by osteoblasts cells and deposited as bone.

Distribution of urinary selenium and arsenic among pregnant women exposed to arsenic in drinking water.

Inorganic arsenic (In-As) is a well-known toxicant and carcinogen found naturally in surface and groundwater around the world. Exposure can cause skin lesions, adverse reproductive outcomes, and cancer. There are two main pathways of arsenic (As) metabolism in humans: the reduction reactions, and the oxidative methylation reactions, where methyl groups are attached to As compounds to form monomethylarsenate (MMA) and dimethylarsenate (DMA). MMA, DMA, and In-As are excreted in urine. Urinary levels of another metalloid, selenium (Se), have recently been shown to be associated with increased As excretion and altered metabolite distribution. This study investigates this association, using data collected in a larger prospective study of arsenic and reproductive effects in Chile. This analysis included 93 pregnant women from Antofagasta. Data on demographic, behavioral, and other characteristics were obtained via interviews conducted by trained midwives, and spot urine samples were analyzed for As and Se concentration using inductively coupled plasma-mass spectrometry (ICP-MS). Urinary Se levels were found to be correlated with urinary As levels in bivariate analysis (r = 0.68, P < 0.01). Multiple linear regression analyses revealed that higher urinary Se levels were associated with increased urinary As excretion, increased %DMA, and decreased %In-As. The results of this study suggest that in populations exposed to arsenic, Se intake may be correlated with urinary As excretion, and may alter As methylation.

Uranium analysis in urine by inductively coupled plasma dynamic reaction cell mass spectrometry.

Urine uranium concentrations are the best biological indicator for identifying exposure to depleted uranium (DU). Internal exposure to DU causes an increased amount of urine uranium and a decreased ratio of 235U/238U in urine samples, resulting in measurements that vary between 0.00725 and 0.002 (i.e., natural and depleted uranium's 235U/238U ratios, respectively). A method based on inductively coupled plasma dynamic reaction cell mass spectrometry (ICP-DRC-MS) was utilized to identify DU in urine by measuring the quantity of total U and the 235U/238U ratio. The quantitative analysis was achieved using 233U as an internal standard. The analysis was performed both with and without the reaction gas oxygen. The reaction gas converted ionized 235U+ and 238U+ into 235UO2+ (m/z = 267) and 238UO2+ (m/z = 270). This conversion was determined to be over 90% efficient. A polyatomic interference at m/z 234.8 was successfully removed from the 235U signal under either DRC operating conditions (with or without oxygen as a reaction gas). The method was validated with 15 urine samples of known uranium compositions. The method detection limit for quantification was determined to be 0.1 pg U mL(-1) urine and an average coefficient of variation (CV) of 1-2% within the sample measurements. The method detection limit for determining 235U/238U ratio was 3.0 pg U mL(-1) urine. An additional 21 patient samples were analyzed with no information about medical history. The measured 235U/238U ratio within the urine samples correctly identified the presence or absence of internal DU exposure in all 21 patients.

Environmental exposure to trace elements and prostate cancer in three New Zealand ethnic groups.

A stratified random sample of 176 men was taken from a larger community prostate study group of 1405 eligible subjects from three ethnic groups in the Wellington region of New Zealand, in order to examine ethnic differences in exposure to cadmium (Cd), selenium (Se) and zinc (Zn) and possible associations of blood levels of Cd, Se and Zn with the prevalence of elevated serum Prostate Specific Antigen (PSA); a marker of prostate cancer. Maori and Pacific Islands men were found likely to have higher Cd exposure than New Zealand Europeans through diet, occupation and smoking. However, there was no significant difference between ethnic groups in mean blood Cd levels. Pacific Islands men had significantly higher levels of blood Se than both New Zealand European men and Maori men. Maori men had significantly higher levels of blood Zn than both New Zealand European men and Pacific Islands men. A positive association was found between blood Cd and total serum PSA. Se and Zn levels were not associated with elevated PSA. Maori and Pacific Islands men have higher prostate cancer mortality rates than New Zealand European men. Ethnic differences in mortality could be contributed to by differences in rates of disease progression, influenced by exposure and/or deficiency to trace elements. However, results did not reflect a consistent ethnic trend and highlight the complexity of the risk/protective mechanisms conferred by exposure factors. Further research is needed to ascertain whether the associations found between Cd and PSA levels are biologically important or are merely factors to be considered when interpreting PSA results clinically.