Analytical Studies of Iodine in Food Substances

Four species of edible brown marine algae were fractionated by trichloroacetic acid precipitation and subsequent chromatography; and the iodine content of each fraction was determined. Quantitative determination of iodide form iodine was made possible by elimination of interfereing substances in cell extract using a weakly basic anion exchange resin column without drastic procedure.

Although the iodine content of each algae was of much diversity owing to species, the iodide form iodine varied between 83~85% of total iodine independent on total iodine content. The rest of the iodine was mostly found in trichloroacetic acid precipitate probably in protein bound form; and each algae contained little trichloroacetic acid soluble non iodide form of iodine, probably a low molecular organic form iodine.     

Iodine distribution in the thyroid follicles of the hagfish,Eptatretus burgeri and lamprey,Lampetra japonica: Electron-probe X-ray microanalysis

Summary In the thyroid follicles of species of cyclostomes, a hagfish and a lamprey, the distribution of stable iodine was examined by electron-probe X-ray microanalysis. A high concentration of stable iodine, heterogeneously distributed, was observed in the follicular cells of hagfish thyroid follicles. In the lamprey a low concentration of iodine was seen in the follicular lumina. The relative values for stable iodine determined in this way corresponded to values obtained by a chemical analytical method.     

Iodine speciation in human serum by reversed-phase liquid chromatography-ICP-mass spectrometry

Reversed-phase liquid chromatography-inductively coupled plasma mass spectrometric hyphenation was used for iodine speciation in human serum. First investigations showed that iodine species nearly quantitatively were eluted in the void volume. The result indicated that protein-linked thryoid hormones were not interacting with the stationary phase, thus being not retained. Investigations were performed about T4-TBG (thyroxin-thyroxin-binding globulin) complex generation and its retention during chromatography. It was shown that T4-TBG was not retained on the column. Therefore, a protease treatment was introduced for serum sample preparation. The analysis of “normal” sera (after protease) gave reasonable results lying in the range published in literature: I-:11; di-iodothyrosine (DIT): 2.1; mono-iodothyrosine (MIT): 1.6; reversed tri-iodothyronine (rT3): 3.9; T3: 5.9; T4: 60; each micrograms iodine per liter. The method also proved to recognize abnormalities in a pathologic serum, having rT3 as the predominant species. In this case the method obviously was superior compared to standard immunoassay methods, as it is monitoring the iodine in the species (physiologically active iodine species), whereas immunoassay methods may sometimes detect deiodinated (inactive) compounds.     

Method developments for iodine speciation by reversed-phase liquid chromatography-ICP-mass spectrometry

Method developments are described for a iodine speciation method. This method is based on a reversed-phase liquid chromatography separation combined with inductively coupled plasma-mass spectrometry detection. Investigations are performed concerning the methanol tolerance/stability of inductively coupled plasma, post-column oxidation/volatilization of iodine from species, influences of buffer pH, and column temperature on separation efficiency. The experiments focused on six iodine species, namely iodine, mono-iodothyrosine, di-iodothyrosine, tri-iodothyronine, reversed tri-iodothyronine, and thyroxin. Detection limits were determined between 0.08 and 1.5 μg/L (iodine related), dependent on species. Appplications on a urine sample were performed, showing iodide as the predominant iodine species.     

Isoaccepting lysine transfer ribonucleic acid species of Pseudomonas aeruginosa.

Pseudomonas aeruginosa tRNA was treated with iodine, CNBr and N-ethylmaleimide, three thionucleotide-specific reagents. Reaction with iodine resulted in extensive loss of acceptor activity by lysine tRNA, glutamic acid tRNA, glutamine tRNA, serine tRNA and tyrosine tRNA. CNBr treatment resulted in high loss of acceptor ability by lysine tRNA, glutamic acid tRNA and glutamine tRNA. Only the acceptor ability of tyrosine tRNA was inhibited up to 66% by N-ethylmaleimide treatment, a reagent specific for 4-thiouridine. By the combined use of benzoylated DEAE-cellulose and DEAE-Sephadex columns, lysine tRNA of Ps. aeruginosa was resolved into two isoaccepting species, a major, tRNA Lys1 and a minor, tRNALys1. Co-chromatography of 14C-labelled tRNALys1 and 3H-labelled tRNALys2 on benzoylated DEAE-cellulose at pH 4.5 gave two distinct, non-superimposable profiles for the two activity peaks, suggesting that they were separate species. The acceptor activity of these two species was inhibited by about 95% by iodine and CNBr. Both the species showed equal response to codons AAA and AAG and also for poly(A) and poly(A1,G1) suggesting that the anticodon of these species was UUU. Chemical modification of these two species by iodine did not inhibit the coding response. The two species of lysine of Ps. aeruginosa are truly redundant in that they are indistinguishable either by chemical modification or by their coding response.     

Increment of Iodine Content in Vegetable Plants by Applying Iodized Fertilizer and the Residual Characteristics of Iodine in Soil

As a new attempt to control iodine deficiency disorder (IDD), we explored a method of iodine supplementation by raising the iodine content in vegetables. When grown in the soil supplemented with iodized fertilizer, the three experimental plant species (cucumber, aubergine, and radish) show increasing iodine levels in both leaf and fruit/rhizome tissues as the iodine content added in soil increases. Excessive iodine added to soil can be toxic to plants, whereas the tolerance limit to excessive iodine varies in the three plant species tested. The migration and volatilization of iodine in soil is correlated with the properties of the soil used. The residual iodine in soil increases as the iodine added to soil increases. The diatomite in the iodized fertilizer helps to increase the durability of the iodized fertilizer. This study potentially provides a safe and organic iodine supplementation method to control IDD.     

Microchemical imaging of iodine distribution in the brown alga Laminaria digitata suggests a new mechanism for its accumulation

Brown algal kelp species are the most efficient iodine accumulators among all living systems, with an average content of 1.0% of dry weight in Laminaria digitata. The iodine distributions in stipe and blade sections from L. digitata were investigated at tissue and subcellular levels. The quantitative tissue mapping of iodine and other trace elements (Cl, K, Ca, Fe, Zn, As and Br) was provided by the proton microprobe with spatial resolutions down to 2 μm. Chemical imaging at a subcellular resolution (below 100 nm) was performed using the secondary ion mass spectrometry microprobe. Sets of samples were prepared by both chemical fixation and cryofixation procedures. The latter prevented the diffusion and the leaching of labile inorganic iodine species, which were estimated at around 95% of the total content by neutron activation analysis. The distribution of iodine clearly shows a huge, decreasing gradient from the meristoderm to the medulla. The contents of iodine reach very high levels in the more external cell layers, up to 191 ± 5 mg g⁻¹ of dry weight in stipe sections. The peripheral tissue is consequently the main storage compartment of iodine. At the subcellular level, iodine is mainly stored in the apoplasm and not in an intracellular compartment as previously proposed. This unexpected distribution may provide an abundant and accessible source of labile iodine species which can be easily remobilized for potential chemical defense and antioxidative activities. According to these imaging data, we proposed new hypotheses for the mechanism of iodine storage in L. digitata tissues. In memory of Dr. Charles Mioskowski, “Miko,” who died on 2 June 2007.     

Preliminary investigation on iodine nutrition in captive giant pandas

Background/ObjectiveThe giant panda belongs to the family Ursidae and, as a species of bear, still retains the simple digestive system of a Carnivoran. However, under the pressure of a specific habitat they had to adapt to a plant mono-diet consisting of bamboo with different species and growth stages around the year. A plant-based diet has relatively low iodine content with risk of iodine deficiency. Furthermore, bamboo contains cyanogenic glycosides releasing cyanide whose detoxification metabolite the thiocyanate acts as antagonist against iodine uptake and storage in the thyroid. To date very little is known about the iodine nutritional status of the giant panda, thus this study was conducted to receive the first information about the iodine nutrition of captive giant panda.Subjects/MethodsHere we investigated the iodine content of bamboo with different plant parts/vegetation stage and species and further compounds of the captive giant panda diet. Next, the urinary iodine (UI) and urinary thiocyanate (UT) levels of infant, sub-adult, adult and geriatric captive giant pandas was measured during the periods when the pandas consume both bamboo leaves- and culm (bamboo leaf-culm stage). Afterwards, the UI of 19 adult giant pandas was measured again for the different iodine intake during bamboo shoot stage. Finally, in this study part also the fecal iodine concentration was analyzed for calculation of total iodine excretion in relation to the iodine intake.ResultsBamboo leaves had the highest iodine content (453 μg/kg dry matter (DM)), followed by the shoots (84 μg/kg DM, p < 0.05), while bamboo culm had the lowest value (12 μg/kg DM, p < 0.05). During bamboo leaf-culm stage, giant pandas of different age groups had different UI and UT levels (p < 0.05). Furthermore, UI and UT were positively correlated among sub-adult, adult and geriatric giant pandas (p < 0.05). In adult giant pandas during bamboo shoot stage, the iodine excretion in feces was not different from that in urine while their total iodine excretion was less than their iodine intake (p < 0.05). Moreover, during bamboo shoot stage, the UI level of adult giant pandas was much lower than noted during bamboo leaf-culm stage (p < 0.05).ConclusionsOur results indicate that UI of captive giant pandas was related to their age as well as to the vegetation stage/part of bamboo they consumed reflecting a different periodic iodine supply. Thiocyanate and fecal excretion should be emphasized when considering the iodine nutrition of giant pandas.     

Relationship between iodination and the polypeptide chain composition of thyroglobulin.

Thyroglobulin was isolated from thyroid glands of normal guinea pigs and from animals treated with thiouracil. These preparations were fractionated by isopyknic centrifugation in RbCl into proteins of varying iodine content. When the disulfide bonds of these protein fractions were reduced and analyzed by polyacrylamide gel electrophoresis in Na dodecyl-SO4, t hree species were observed with molecular weights of 295,000 (A), 210,000 (B), and 110,000 (C). Species A comprised 80% of the protein in thyroglobulin of 0.04% iodine and 13% in thyroglobulin of 0.68% iodine content. Species C showed the opposite relationship, comprising 10% of the low and 70% of the high iodine thyroglobulin. Species B was relatively independent of the iodine content and represented approximately 20% of the protein. Iodine analysis of these proteins showed species A to be lowest and species C highest. It appears that the subunit composition of thyroglobulin depends on the degree of iodination and that species A should be the only one present in the absence of iodination.     

Presence and Function of Sulfur-containing Transfer Ribonucleic Acid of Bacillus subtilis

Bacillus subtilis transfer ribonucleic acid (tRNA) was analyzed for the occurrence of thionucleotides by in vivo labeling with (35)S and fractionation by methylated albumin kieselguhr column chromatography. Alkaline hydrolysates of tRNA were also examined by column chromatography and paper electrophoresis, and the amino acid-accepting ability of thionucleotide-containing tRNA was tested after iodine oxidation. The results showed that B. subtilis tRNA contains 4-thiouridylate, a second nucleotide with properties similar to 2-thiopyrimidine, and a third unidentified thionucleotide. The amino acid-accepting ability for serine, tyrosine, lysine, and glutamic acid was markedly inhibited after oxidation of the tRNA with iodine, suggesting the presence of thionucleotides in these tRNA species. This inhibition could be reversed by thiosulfate reduction. The iodine treatment totally inactivated all lysine tRNA species, partially inactivated the serine tRNA species, and did not affect the accepting ability for valine. A comparison of tRNA from cells in the log and stationary phases and from spores revealed similar iodine inactivation patterns in all cases. The thionucleotide content in B. subtilis tRNA differed from that in Escherichia coli, both in extent and in distribution. A possible function of the thionucleotides in tRNA is discussed.     

Effects of cold (+4°C) on thyroid metabolism of iodine (I and I) in quail (Coturnix coturnix japonica)—A comparison with the rat

1. 1.|A kinetic study of the absolute specific activity (SA) of iodine in intrathyroidal iodotyrosines and iodothyronines, and in plasma PBI, was performed comparatively in adult male rats and quail kept at +25°C (controls) or exposed to +4°C (for 2 days or 30 days before the iodine measurements).

2. 2.|In both species, 2 days of cold exposure induced a rapid increase (20 to 50%) in the SA of intrathyroidal iodoaminoacids and plasma PBI, leading to a rapid equilibration between the SA measured. After 30 days an acceleration in iodine turnover was still measurable in rats, but the stimulation then appeared more important in quail. In addition, the acceleration of iodine turnover led to a characteristic bimodal pattern of this turnover which was apparent only in quail, and could reflect an important production of non-hormonal iodinated proteins. Although the bird thyroid has been claimed to be less reactive to cold than its mammalian homologues, the present study indicates that, at least in bird species whose thyroidal stores of iodine are not excessive, the thyroid is even more responsive to cold than the rat thyroid.

Reduction of iodate in iodated salt to iodide during cooking with iodine as measured by an improved HPLC/ICP–MS method

Background: Iodate is a strong oxidant, and some animal studies indicate that iodate intake may cause adverse effects. A key focus of the safety assessment of potassium iodate as a salt additive is determining whether iodate is safely reduced to iodide in food. Objective: To study the reduction of iodate in table salt to iodide and molecular iodine during cooking. Materials and Methods: Fifteen food samples cooked with and without iodated salt were prepared in duplicate. The iodine in the cooked food was extracted with deionized water. The iodine species in the extracts were determined by using an improved high-performance liquid chromatography/inductively coupled plasma–mass spectrometry (HPLC/ICP–MS). The cooking temperature and the pH of the food were determined. Results: The conversion rate of iodate in iodated salt to iodide and molecular iodine was 96.4%±14.7% during cooking, with 86.8%±14.5% of the iodate converted to iodide ions and 9.6% ±6.2% converted to molecular iodine to lose. The limit of detection, limit of quantification, relative standard deviation and recovery rate of the method HPLC/ICP–MS were 0.70 μg/L for I⁻ (0.69 μg/L for IO₃⁻), 2.10 μg/L for I⁻ (2.06 μg/L for IO₃⁻), 2.6% and 101.6%±2.6%, respectively. Conclusion: Almost all iodate added to food was converted into iodide and molecular iodine during cooking. The improved HPLC/ICP–MS was reliable in the determination of iodine species in food extracts.     

Uptake of Different Species of Iodine by Water Spinach and Its Effect to Growth

A hydroponic experiment has been carried out to study the influence of iodine species [iodide (I(-)), iodate ([Formula: see text]), and iodoacetic acid (CH(2)ICOO(-))] and concentrations on iodine uptake by water spinach. Results show that low levels of iodine in the nutrient solution can effectively stimulate the growth of biomass of water spinach. When iodine levels in the nutrient solution are from 0 to 1.0 mg/l, increases in iodine levels can linearly augment iodine uptake rate by the leafy vegetables from all three species of iodine, and the uptake effects are in the following order: [Formula: see text]. In addition, linear correlation was observed between iodine content in the roots and shoots of water spinach, and their proportion is 1:1. By uptake of I(-), vitamin C (Vit C) content in water spinach increased, whereas uptake of [Formula: see text] and CH(2)ICOO(-) decreased water spinach Vit C content. Furthermore, through uptake of I(-) and [Formula: see text], the nitrate content in water spinach was increased by different degrees.     

A newly discovered oxidant defence system and its involvement in the development of Aurelia aurita (Scyphozoa, Cnidaria): reactive oxygen species and elemental iodine control medusa formation

In Aurelia aurita, applied iodine induces medusa formation (strobilation). This process also occurs when the temperature is lowered. This was found to increase oxidative stress resulting in an increased production of iodine from iodide. One polyp produces several medusae (initially termed ephyrae) starting at the polyp's oral end. The spreading of strobilation down the body column is controlled by a feedback loop: ephyra anlagen decrease the tyrosine content in adjacent polyp tissue by producing melanin from tyrosine. Endogenous tyrosine is able to remove iodine by forming iodiferous tyrosine compounds. The reduced level of tyrosine causes the ephyra-polyp-border to move towards the basal end of the former polyp. We argue that an oxidant defence system may exist which makes use of iodide and tyrosine. Like other marine invertebrates, polyps of Aurelia contain iodide ions. Inevitably produced peroxides oxidise iodide into iodine. The danger to be harmed by iodine is strongly decreased by endogenous tyrosine which reacts with iodine to form iodiferous tyrosine compounds including thyroxin. Both substances together, iodide and tyrosine, form an efficient oxidant defence system which shields the tissue against damage by reactive oxygen species. In the course of evolution (from a species at the basis of the animal kingdom like Aurelia to a highly evolved species like man) the waste product thyroxin (indicating a high metabolic rate) has developed into a hormone which controls the metabolic rate.     

Availability of iodide and iodate to spinach (Spinacia oleracea L.) in relation to total iodine in soil solution

A greenhouse pot experiment was carried out to investigate the availability of iodide and iodate to soil-grown spinach (Spinacia oleracea L.) in relation to total iodine concentration in soil solution. Four iodine concentrations (0, 0.5, 1, 2 mg kg⁻¹) for iodide (I⁻) and iodate (IO₃⁻) were used. Results showed that the biomass productions of spinach were not significantly affected by the addition of iodate and iodide to the soil, and that iodine concentrations in spinach plants on the basis of fresh weights increased with increasing addition of iodine. Iodine concentrations in tissues were much greater for plants grown with iodate than with iodide. In contrast to the iodide treatments, in iodate treatment leaves accounted for a larger fraction of the total plant iodine. The soil-to-leaf transfer factors (TF_leaf) for plants grown with iodate were about tenfold higher than those grown with iodide. Iodine concentrations in soil solution increased with increasing iodine additions to the soil irrespective of iodine species. However, total iodine in soil solution was generally higher for iodate treatments than iodide both in pots with and without spinach. According to these results, iodate can be considered as potential iodine fertilizer to increase iodine content in vegetables.     

Effects of lodine in Various Formulations on the Growth of Barley and Pea Plants in Nutrient Solution Culture

The forms of iodine added to cultures of barley were potassiumiodide, potassium iodate, potassium periodate, and iodoaceticacid at iodine concentrations of 1.0 ppm and 10.0 ppm. Withpea, only iodide and iodate at 1.0 ppm iodine concentrationwere used. For both species, comparisons were made with culturesto which no iodine was added. In barley, growth was increased by 1.0 ppm iodine, the relativeeffectiveness of the different formulations being in the order:iodoacetic acid > iodide > iodate > periodate. With10.0 ppm, iodide and iodoacetic acid treatments gave reducedgrowth, iodate was without effect, and periodate enhanced growth. In pea, 1.0 ppm iodine was inhibitory, iodide being more toxicthan iodate. Analysis of dry matter showed iodine content according to treatmentto be in the order: iodide > iodoacetic acid > iodate> periodate     

Insulin amyloid fibrils form an inclusion complex with molecular iodine: a misfolded protein as a nanoscale scaffold

The study describes formation of an intensely violet inclusion complex of insulin amyloid fibrils and molecular iodine. Resonance Raman spectra of complexes formed by staining mature insulin fibrils with iodine and by seeding fibrils in the presence of iodine imply similar topologies of entrapped iodine and oligoiodide species. Iodine captured by growing fibrils remains accessible to a bulk chemical reagent. In light of its small size and the fact that iodine can partition into polar as well as nonpolar media, the data suggest that intrafibrillar structure of insulin amyloid is densely packed with no appreciable void volumes capable of accommodating iodine atoms. The complex is stable: only drastic perturbation of the beta-pleated fibrous scaffold by dimethyl sulfoxide (rather than of the beta-sheet conformation) leads to the release of iodine atoms from surface moieties. While the reaction between iodine and in vivo amyloid deposits was first described by Virchow in the 19th Century [Virchow, R. (1854) Virchows Arch. 6, 268-271], the underlying molecular mechanism has not been thoroughly explored since then. This work shows how the long-forgotten concept can be utilized as a probe of void volumes in protein fibrils, providing a new tool for structural studies on amyloids, and a model for design of protein-based drug delivery media.     

Study on chemical species of iodine in human liver

The distribution and chemical species of iodine in various subcellular fractions of human liver were studied by using epithermal neutron activation analysis combined with chemical and biochemical separation techniques, such as gradient centrifugation and gel chromatography. It was found that the total iodine content orders in various subcellular fractions is as follows: nuclei > cytosol > mitochondria > lysosome > microsome. In the lysosomal fraction, iodine is mainly bound to macromolecules, whereas in the nuclei and mitochondrial fractions, mainly with lower-molecular-weight organic compounds. In the cytosol fraction, iodine is combined with three proteins, in which iodine is chiefly bound with mid- and high-molecular-weight proteins.     

The effects of some halogen-containing compounds on Bacillus subtilis endospores

Sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were more active against Bacillus subtilis 8236 spores in both viability and in germination and outgrowth studies than were polyvinylpyrrolidone-iodine (PVP-I) and Lugol's solution. Of the two chlorine compounds studied NaOCl proved to be the more active. The two iodine-containing compounds gave contrasting results with the Lugol's solution demonstrating increased antibacterial activity with increasing available iodine concentration. The antibacterial behaviour of PVP-I, however, reflected the more complex nature of aqueous iodine-surfactant mixtures. Initially, non-complexed iodine concentration (the active species) increased with increasing total available iodine concentration, resulting in increasing antibacterial activity. However, due to changes in the physical properties of the mixture, a maximum concentration of non-complexed iodine was reached so that a further increase in total available iodine resulted in a decrease in non-complexed iodine concentration and consequently a decrease in the antibacterial activity of the solution was observed. A greater inhibitory effect was observed in subsequent germination and outgrowth studies when spores were pre-treated with respective biocide than when untreated spores were added to germination media containing biocide at t = 0. This may reflect a combination of different contact times plus the neutralizing effect of the germination media on such halogen compounds.     

The effects of some halogen-containing compounds on Bacillus subtilis endospores

Sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were more active against Bacillus subtilis 8236 spores in both viability and in germination and outgrowth studies than were polyvinylpyrrolidone-iodine (PVP-I) and Lugol's solution. Of the two chlorine compounds studied NaOCl proved to be the more active. The two iodine-containing compounds gave contrasting results with the Lugol's solution demonstrating increased antibacterial activity with increasing available iodine concentration. The antibacterial behaviour of PVP-I, however, reflected the more complex nature of aqueous iodine—surfactant mixtures. Initially, non-complexed iodine concentration (the active species) increased with increasing total available iodine concentration, resulting in increasing antibacterial activity. However, due to changes in the physical properties of the mixture, a maximum concentration of non-complexed iodine was reached so that a further increase in total available iodine resulted in a decrease in non-complexed iodine concentration and consequently a decrease in the antibacterial activity of the solution was observed.
A greater inhibitory effect was observed in subsequent germination and outgrowth studies when spores were pre-treated with respective biocide than when untreated spores were added to germination media containing biocide at t = 0. This may reflect a combination of different contact times plus the neutralizing effect of the germination media on such halogen compounds.