Illicium verum (Star Anise) and Trans-Anethole as Valuable Raw Materials for Medicinal and Cosmetic Applications.

Illicium verum Hook f. (star anise) is considered an important species in Traditional Chinese Medicine and is also used in contemporary medicine in East Asian countries. It occurs in natural habitats in southeastern parts of China and Vietnam, and is cultivated in various regions in China. The raw materials-Anisi stellati fructus and Anisi stellati aetheroleum obtained from this species exhibit expectorant and spasmolytic activities. The European Pharmacopoeia (4th edition) indicates that these raw materials have been used in allopathy since 2002. The biological activities of the above-mentioned raw materials are determined by the presence of valuable secondary metabolites such as monoterpenoids, sesquiterpenoids, phenylpropanoids, and flavonoids. Recent pharmacological studies on fruit extracts and the essential oil of this species have confirmed their antibacterial, antifungal, anti-inflammatory, and antioxidant activities and thus their medicinal and cosmetic value. The aim of this review was to examine the progress of phytochemical and pharmacological studies that focused on possible cosmetic applications. In addition to fruit extracts and essential oil, the current consensus on the safety of trans-anethole, which is the main compound of essential oil used in cosmetology, is underlined here.

The effect of tree age, daily sap volume and date of sap collection on the content of minerals and heavy metals in silver birch (Betula pendula Roth) tree sap.

The aim of this study was to determine the effect of the age of trees, daily sap volume as well as the term of tapping birch sap collected in the forest environment on the content of selected minerals (zinc, copper and manganese) and heavy metals (lead, nickel, chromium and cadmium). The study was performed on material taken from two stands (aged 34 and 84 years) in a moist broadleaved forest habitat with a dominant share of silver birch (Betula pendula Roth). The research results confirmed the presence of both nutritional essential minerals and hazardous heavy metals in the birch sap. At the same time, the content of minerals and heavy metals was found to be very variable and the differences between their concentrations, recorded on the same day of collecting in several trees of the same age group, can be even several dozen times higher. Depending on the examined elements, the factors influencing their content vary. The age of the trees determines only the manganese content; daily sap volume significantly affects the content of manganese and copper, and date of collection differentiates the content of zinc, lead, nickel and cadmium. The results may be interesting in the context of developing procedures for collecting birch sap for the purpose of obtaining raw material with beneficial nutritional values and a high level of health safety. For this reason, our recommendation for guaranteeing the health safety and high nutritional value of birch sap is to combine batches of raw material taken from as many trees as possible, and at the same time to publicize the fact that collecting birch sap from just one single tree may result in a raw material that is both dangerous and has no nutritional benefits.

Polyaniline based material as a new SPE sorbent for pre-treatment of Chelidonium majus extracts before chromatographic analysis of alkaloids.

In the present study, deprotonated and protonated polyaniline based sorbent obtained by in situ polymerization of aniline on silica (Si-PANI) was applied for SPE pretreatment of Chelidonium majus extracts before determination of benzophenanthridine, protoberberine and protopine alkaloids. The experimentally optimized conditions, such as protonated polyaniline impregnated with methanol as a sorbent, 2 mL of water and methanol mixture (1/1, v/v) as a washing solution, and 5 mL of 0.1 M methanolic solution of ammonia as an elution solvent, yielded the highest values of analyte recovery (above 96.5%) with simultaneous removal of undesirable plant matrix. Virtually modeled structure of polyaniline revealed possible occurrence of the mixed mode binding mechanism, based both on π-π and ion interactions on protonated form of polyaniline. Moreover, lack of significant changes of the polyaniline film quality assessed by Raman spectroscopy after series of ten experiments proved the reusability of the sorbent.

Methodological approach to determine carlina oxide - a main volatile constituent of Carlina acaulis L. essential oil.

In this work, a fast and low-cost voltammetric methodology for determination of carlina oxide in plant extracts was developed. The best results were obtained using a boron-doped diamond electrode (BDDE). The voltammetric measurements of carlina oxide were performed in a 0.1 mol/L solution of sulphuric acid. After 30 s of stirring the solution, differential pulse voltammograms (DPVs) were recorded from 0.5 to 1.8 V. The amplitude was 75 mV and the scan rate was 175 mV/s. Measurements were recorded in non-deaerated solutions. The background current was subtracted from each registered voltammogram; then they were cut from 0.5 to 1.5 V. The detection and quantification limits were 0.28 and 0.93 µg/L, respectively, and repeatability expressed as the relative standard deviation of 0.1 mg/L of carlina oxide was 1.9% (n = 5). The results were compared with those obtained using gas chromatography with a flame ionization detector and high performance liquid chromatography with a photodiode array detector.

Systematic Evaluation of Chromatographic Parameters for Isoquinoline Alkaloids on XB-C18 Core-Shell Column Using Different Mobile Phase Compositions.

Chelidonium majus L. is a rich source of isoquinoline alkaloids with confirmed anti-inflammatory, choleretic, spasmolytic, antitumor, and antimicrobial activities. However, their chromatographic analysis is difficult because they may exist both in charged and uncharged forms and may result in the irregular peak shape and the decrease in chromatographic system efficacy. In the present work, the separation of main C. majus alkaloids was optimized using a new-generation XB-C18 endcapped core-shell column dedicated for analysis of alkaline compounds. The influence of organic modifier concentration, addition of salts, and pH of eluents on chromatographic parameters such as retention, resolution, chromatographic plate numbers, and peak asymmetry was investigated. The results were applied to elaborate the optimal chromatographic system for simultaneous quantification of seven alkaloids from the root, herb, and fruit of C. majus.

Proliferative and antioxidant activity of Symphytum officinale root extract.

The root of Symphytum officinale L. is commonly used in folk medicine to promote the wound healing, reduce the inflammation and in the treatment of broken bones. The objective of our investigation was to analyse the extract from S. officinale in term of its antioxidant activity and the effect on cell viability and proliferation of human skin fibroblast (HSF). Moreover, the quantification of main phenolics and allantoin was conducted using HPLC-DAD method. Five compounds were found: rosmarinic, p-hydroxybenzoic, caffeic, chlorogenic and p-coumaric acid. DPPH, FRAP and TPC assay showed the high antioxidant activity of the extract. MTT test proved the stimulatory effect on cell metabolism and viability of HSF cells. Moreover, no changes in cytoskeleton structure and cells shape were observed. The obtained results indicate that non-toxic extract from S. officinale root has strong antioxidant potential and a beneficial effect on human skin fibroblasts.

Carlina vulgaris L. as a Source of Phytochemicals with Antioxidant Activity.

The methanol extracts from three populations of Carlina vulgaris L. were examined for the chlorogenic acid content, mineral content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity. Two populations originated from natural nonmetallicolous habitats (NN (populations from Nasilów) and NP (populations from Pinczów)), and one metallicolous population (MB) was collected from Boleslaw waste heap localized at the place of former open-cast mining of Ag-Pb and Zn-Pb ores dating back to the 13th century and 18th century, respectively. The level of Zn, Pb, Cd, Fe, Ni, and Mn was significantly higher in the root and leaves of MB plants as a result of soil contaminations compared to those of the NN and NP ones. The highest antioxidant potency has been showed by the plants growing in a nonmetallicolous habitat. The flower head extracts obtained from the nonmetallicolous populations also contained the largest amount of chlorogenic acid, whereas the lowest was determined in the roots (ca. 2-3.5 mg/g and 0.2-0.4 mg/g of air-dry weight, resp.). These studies provide important information on the influence of a habitat on the quality of herbal materials and the content of the biologically active primary and secondary metabolites.

Application of Raman spectroscopy for direct analysis of Carlina acanthifolia subsp. utzka root essential oil.

Carlina genus plants e.g. Carlina acanthifolia subsp. utzka have been still used in folk medicine of many European countries and its biological activity is mostly associated with root essential oils. In the present paper, Raman spectroscopy (RS) was applied for the first time for evaluation of essential oil distribution in root of C. acnthifolia subsp. utzka and identification of root structures containing the essential oil. Furthermore, RS technique was applied to assess chemical stability of oil during drying of plant material or distillation process. Gas chromatography-mass spectrometry was used for qualitative and quantitative analysis of the essential oil. The identity of compounds was confirmed using Raman, ATR-IR and NMR spectroscopy. Carlina oxide was found to be the main component of the oil (98.96% ± 0.15). The spectroscopic study showed the high stability of essential oil and Raman distribution analysis indicated that the oil reservoirs were localized mostly in the structures of outer layer of the root while the inner part showed nearly no signal assigned to the oil. Raman spectroscopy technique enabled rapid, non-destructive direct analysis of plant material with minimal sample preparation and allowed straightforward, unambiguous identification of the essential oil in the sample.

In Vitro Antiproliferative Activity of Extracts of Carlina acaulis subsp. caulescens and Carlina acanthifolia subsp. utzka.

Various species of the Carlina genus have been used in traditional medicine in many countries to treat numerous skin disorders, including cancer. The objective of this work was to assess the anticancer properties of root and leaf extracts from Carlina acaulis subsp. caulescens and C. acanthifolia subsp. utzka. Anti-tumor properties of the extracts were explored using a tetrazolium-based cell viability assay and flow cytometric apoptosis analysis, followed by immunodetection of phosphoactive ERK1/2 in UACC-903, C32, and UACC-647 human melanoma cell lines. Normal human fibroblasts were used as a control. Leaf extracts inhibited the viability of all tested melanoma cell lines in a dose-dependent fashion while the fibroblasts were less sensitive to such extract. The root extracts inhibited the proliferation of UACC-903 and UACC-647 cells only at the highest doses (300 µg/mL). However, the C32 and fibroblast cells exhibited an increase in the cellular proliferation rate and no caspase activity was observed in response to the root extracts (100 µg/mL). An increase in caspase activity was observed in melanoma cells treated with the leaf extracts of both Carlina species. Leaf extracts from C. acaulis subsp. caulescens (100 µg/mL) inhibited proliferatory ERK1/2 in UACC-903 and C32 cells, as demonstrated by the decrease in ERK1/2 phosphorylation. No reduction in phospho-ERK1/2 was observed in the tested cell lines treated with the root extracts, apart from UACC-647 after incubation with the C. acanthifolia subsp. utzka root extract (100 µg/mL). There was no change in ERK1/2 phosphorylation in the fibroblasts. The extracts from the leaves and roots were analyzed by HPLC and the analysis showed the presence of triterpenes and phenolic acids as the main extract components. The research demonstrated that the extracts from the leaves of the plants were cytotoxic against the human melanoma line and induced apoptosis of the cells. The triterpene fraction present in the tested extracts may be responsible for this activity.

Alterations of Hair and Nail Content of Selected Trace Elements in Nonoccupationally Exposed Patients with Chronic Depression from Different Geographical Regions.

The aim of this study was to determine if altered levels of selected trace elements manifest themselves during chronic depression. To identify elements strongly associated with chronic depression, relationships between the elemental contents of hair and nails and the interelement correlations were checked. Inductively coupled plasma mass spectrometry and ion chromatography were used to evaluate the contents of Zn, Cu, Co, Pb, Mn, and Fe in hair and nail samples from a total of 415 subjects (295 patients and 120 healthy volunteers). The study included logistic regression models to predict the probability of chronic depression. To investigate possible intercorrelations among the studied elements, the scaled principal component analysis was used. The research has revealed differences in TE levels in the group of depressed men and women in comparison to the healthy subjects. Statistically significant differences in both hair and nails contents of several elements were observed. Our study also provides strong evidence that the intermediary metabolism of certain elements is age- and gender-dependent. Zn, Mn, Pb, and Fe contents in hair/nails seem to be strongly associated with chronic depression. We found no statistically significant residence-related differences in the contents of studied elements in nonoccupationally exposed patients and healthy subjects.

Optimization of high-resolution continuum source graphite furnace atomic absorption spectrometry for direct analysis of selected trace elements in whole blood samples.

Trace analysis plays an important role in medicine for diagnosis of various disorders; however, the appropriate sample preparation is required mostly including mineralization. Although graphite furnace atomic absorption spectrometry (GF AAS) allows the investigation of biological samples such as blood, serum, and plasma without this step, it is rarely used for direct analysis because the residues of the rich organic matrix inside the furnace are difficult to remove and this may cause spectral/matrix interferences and decrease the lifetime of the graphite tube. In our work, the procedure for determination of Se, Cr, Mn, Co, Ni, Cd and Pb with the use of the high resolution continuum source GF-AAS technique in whole blood samples with minimum sample pre-treatment was elaborated. The pyrolysis and atomization temperature as well as the time of signal integration were optimized to obtain the highest intensity and repeatability of the analytical signal. Moreover, due to the apparatus modification, an additional step was added in the for graphite furnace temperature program with minimal argon flow and maximal flow of air during pyrolysis stage to increase the oxidative condition for better matrix removal. The accuracy and precision of the optimized method was verified using certified reference material (CRM) Seronorm Trace Elements Whole Blood L-1 and the developed method was applied for trace analysis of blood samples from volunteer patients of the Orthopedics Department.

Evaluation of seasonal changes of triterpenic acid contents in Viscum album from different host trees.

CONTEXT: Viscum album L. (Loranthaceae) is a semi-parasitic plant used in pharmacy and medicine mostly for its hypotensive and anticancer activity. The effects may be related to the presence of triterpenic acids, such as betulinic (BA) and oleanolic (OA) acids. OBJECTIVES: In our investigations the content of triterpenic acids in V. album from different host trees depending on the season of harvest was determined. MATERIAL AND METHODS: V. album herb was dried and extracted with ethyl acetate using ultrasound energy. The reversed phase HPLC-PDA method was used for the analysis of triterpenic acids. The structure of the target components was confirmed by mass spectrometry with an electrospray ionization source. RESULTS: Diversity in the content of both compounds was noted; however, OA was the dominant triterpenic acid and the amount thereof was ∼10 times higher than that of BA. The analysis of changes in the amount of triterpenic acids during the spring-winter period revealed the highest content of OA in summer (from 6.84 to 13.65 mg/g). In turn, in the other seasons of harvest, the content was in the range of 4.41-9.83, 6.41-9.56 and 5.59-12.16 mg/g for spring, autumn and winter, respectively. In most cases, a similar tendency was observed for BA. DISCUSSION AND CONCLUSION: In most cases, the highest amount of the investigated compounds was found in summer; thus, this period seems to be optimal for acquisition of plant material rich in triterpenic acids.

Iodine in autism spectrum disorders.

OBJECTIVE: The aim of our study was to assess the iodine status of Polish boys with severe autism compared to their healthy peers and evaluate the relationship between urinary iodine, thyroid hormones, body mass index and Autism Spectrum Disorder (ASD) symptomatology. SUBJECTS AND METHODS: Tests were performed in 40 boys with ASD and 40 healthy boys, aged 2-17 from the same geographic region in Poland. Urinary iodine (UI), free triiodothyronine (fT3), free thyroxine (fT4), thyroid stimulating hormone (TSH), BMI, and individual symptoms measured by the Childhood Autism Rating Scale (CARS) were correlated. Validated ion chromatography method with pulsed amperometric detection was applied for the determination of urinary iodine after optimized alkaline digestion in a closed system assisted with microwaves. RESULTS: 19 out of 40 children with ASD had mild to moderate iodine deficiency. Statistically significant lower levels of UI, fT3 and fT4 and higher levels of TSH were found in the autistic group when compared with the control group. Concentration of iodine in urine was negatively associated with clinician's general impression for children between 11 and 17 years. Emotional response, adaptation to environmental change, near receptor responsiveness, verbal communication, activity level, and intellectual functioning are more associated with UI than other symptoms listed in CARS. CONCLUSION: The severity of certain symptoms in autism is associated with iodine status in maturing boys. Thyroid hormones were within normal reference ranges in both groups while urinary iodine was significantly lower in autistic boys suggesting that further studies into the nonhormonal role of iodine in autism are required.

The Stimulatory Effect of Strontium Ions on Phytoestrogens Content in Glycine max (L.) Merr.

The amount of secondary metabolites in plants can be enhanced or reduced by various external factors. In this study, the effect of strontium ions on the production of phytoestrogens in soybeans was investigated. The plants were treated with Hoagland's solution, modified with Sr(2+) with concentrations ranging from 0.5 to 3.0 mM, and were grown for 14 days in hydroponic cultivation. After harvest, soybean plants were separated into roots and shoots, dried, and pulverized. The plant material was extracted with methanol and hydrolyzed. Phytoestrogens were quantified by HPLC. The significant increase in the concentration of the compounds of interest was observed for all tested concentrations of strontium ions when compared to control. Sr(2+) at a concentration of 2 mM was the strongest elicitor, and the amount of phytoestrogens in plant increased ca. 2.70, 1.92, 3.77 and 2.88-fold, for daidzein, coumestrol, genistein and formononetin, respectively. Moreover, no cytotoxic effects were observed in HepG2 liver cell models after treatment with extracts from 2 mM Sr(2+)-stressed soybean plants when compared to extracts from non-stressed plants. Our results indicate that the addition of strontium ions to the culture media may be used to functionalize soybean plants with enhanced phytoestrogen content.

Adsorption Kinetics at Silica Gel/Ionic Liquid Solution Interface.

A series of imidazolium and pyridinium ionic liquids with different anions (Cl(-), Br(-), BF4(-), PF6(-)) has been evaluated for their adsorption activity on silica gel. Quantification of the ionic liquids has been performed by the use of RP-HPLC with organic-aqueous eluents containing an acidic buffer and a chaotropic salt. Pseudo-second order kinetic models were applied to the experimental data in order to investigate the kinetics of the adsorption process. The experimental data showed good fitting with this model, confirmed by considerably high correlation coefficients. The adsorption kinetic parameters were determined and analyzed. The relative error between the calculated and experimental amount of ionic liquid adsorbed at equilibrium was within 7%. The effect of various factors such as initial ionic liquid concentration, temperature, kind of solvent, kind of ionic liquid anion and cation on adsorption efficiency were all examined in a lab-scale study. Consequently, silica gel showed better adsorptive characteristics for imidazolium-based ionic liquids with chaotropic anions from aqueous solutions in comparison to pyridinium ionic liquids. The adsorption was found to decrease with the addition of organic solvents (methanol, acetonitrile) but it was not sensitive to the change of temperature in the range of 5-40 °C.

Concentration of Selected Metals in Whole Blood, Plasma, and Urine in Short Stature and Healthy Children.

The short stature in children is defined as height below the third percentile from the mean for age and gender. This problem affects about 3% of young people. More than 20,000 children in Poland have problems with short stature. There is not much information available in the literature on the study of metals in blood, plasma, and urine in children with short stature. The study was conducted on a group of 56 short stature Polish children and 35 healthy children. The content of metals was determined using high-performance ion chromatography and inductively coupled plasma mass spectrometry methods. The study revealed significant differences between the content of selected metals in body fluids between a short stature group and healthy children. There were significant differences in the Fe, Cu, and Ni concentrations between the groups with respect to the hormonal therapy. There were no significant differences between the groups with respect to the area where the children lived. The results showed no statistically significant differences between metal concentration and age, body weight, and height. The study demonstrated statistically significant differences between the content of metals in body fluids in short stature children compared with the healthy children. It seems that the difference in the concentration of certain elements may also be the result of growth hormone therapy and the interaction between various metals. Both the alterations in the content of metals and their mutual interactions may play an important role in the pathogenesis of short stature children.

Serum and urinary selenium levels in obese children: a cross-sectional study.

OBJECTIVE: To determine serum and urinary selenium (Se) levels in children with and without obesity, and to assess if Se influences the risk of obesity. SUBJECTS AND METHODS: High-resolution-continuum source-atomic absorption spectrometry (HR-CS-AAS) was used to determine the content of Se in 80 children (age 6-17; 40 boys, 40 girls). Correlations between variables were tested with the use of Spearman's correlation coefficient. U Mann-Whitney test was applied to assess the difference of Se contents in samples. Measured metabolic risk factors (blood pressure, glucose level, triglycerides (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and total cholesterol), age, gender, and BMI were correlated. Logistic regression models were fitted to identify predictors of obesity interacting with selenium content in serum and urine, separately. RESULTS: Obese children, regardless of gender, had lower Se content. Se level in serum (p=0.001, OR 0.74, 95%CI 0.62-0.88) and total cholesterol (p=0.001, OR 1.19, 95%CI 1.08-1.31) were the independent factors significantly influencing the risk of obesity in children. Two separate models were observed for Se in urine: (i) Se level (p<0. 0001, OR 0.70, 95%CI 0.58-0.84) and glucose level (p<0.0001, OR 1.22, 95%CI 1.10-1.35), and (ii) Se level (p=0.002, OR 0.60 95%CI 0.43-0.83) and total cholesterol level (p=0.003, OR 1.16, 95%CI 1.05-1.28). CONCLUSION: The current study suggests a possible role of Se in obesity. Further research needs to be performed to check if obese children are an at-risk group for Se deficiency.

Herbal preparation extract for skin after radiotherapy treatment. Part One--Preclinical tests.

Naran R is a herbal composition made of Plantago lanceolate folium, Malvae arboreae flos, Calendulae flos, Chamomillae inflorescentia, Lamii albi flos to prepare compresses or to wash skin with inflammations. The extract of this preparation is mixed to be applied as an ointment on patients' skin after radiotherapy. Experiments performed in vitro are part of pre-clinical tests with Naran R ointment. This study examined the impact of the plant composition for ethanol-water extract on human skin fibroblasts (HSF) culture. Samples of extract, prepared from patented amounts of herbs, were in the range of 25-225 µg/mL. Six methods were applied: standard spectrophotometric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, neutral red (NR) uptake assay, DPPH free radical scavenging test, labeling of cytoskeleton F-actin, staining of argyrophilic nucleolar organizer regions (AgNORs) and trypan blue coloration. The extract concentration 75 µg/mL was established as safe for application on human skin. In labeling of F-actin with rhodamine-phalloidin dye at this concentration the cytoskeleton was stable. The extract did not influence the membrane stability and had positive influence on the proliferation activity. It was confirmed in AgNOR test during incubation with extract, which led to formation of larger amount of smaller nucleolins. In DPPH scavenging activity test, the extract revealed over 8% higher free-radical scavenging activity in comparison to control. After trypan blue staining, the extract in concentration 125 µg/mL significantly lowered the cell viability. When the cytotoxic and anti-proliferative activity of the extracts were analyzed, MTT and Neutral Red (NR) methods were used. The cells' viability was maintained on a constant level (80-110%) after 24, 48 and 72 h of incubation. During all time of NR test (72 h) and even when 225 µg/mL of extract was applied, the viability of cells was in range 80-110% of control. Positive influence of the extract on investigated cells structure and proliferation, lack of toxicity and increasing anti-oxidant activity enable to consider this preparation as a natural remedy with potential application in skin therapy after radiation.

Analysis of compounds with phytoestrogenic activity in dietary supplements with use of HPTLC-densitometry method.

Soy (Glycine max L., Fabaceae) and soy products are becoming more popular because of their low toxicity and therapeutic effects. Soy possesses antioxidant, anti-inflammatory and anti-allergic properties, however, the most important is its estrogenic activity associated with occurrence of phytoestrogens. Isoflavones with phytoestrogenic effects were determined in four commercially available soya formulations. Analyses were performed with the use of high performance thin-layer chromatography (HPTLC) combined with densitometry. The compounds were extracted, hydrolyzed in order to obtain aglycone forms and separated on HPTLC silica gel 60 F254 plates with the use of mobile phase consisting of chloroform--ethyl acetate--formic acid 4:6:0.1 (v/v/v). After drying, the spots on the plates were determined in absorbance/reflectance mode at a wavelength of 260 nm using a computer-controlled densitometer Desaga CD 60.

Determination of total iodine in serum and urine samples by ion chromatography with pulsed amperometric detection - studies on analyte loss, optimization of sample preparation procedures, and validation of analytical method.

A fast, accurate and precise ion chromatography method with pulsed amperometric detection was applied to evaluate a variety of parameters affecting the determination of total iodine in serum and urine of 81 subjects, including 56 obese and 25 healthy Polish children. The sample pretreatment methods were carried out in a closed system and with the assistance of microwaves. Both alkaline and acidic digestion procedures were developed and optimized to find the simplest combination of reagents and the appropriate parameters for digestion that would allow for the fastest, least time consuming and most cost-effective way of analysis. A good correlation between the certified and the measured concentrations was achieved. The best recoveries (96.8% for urine and 98.8% for serum samples) were achieved using 1ml of 25% tetramethylammonium hydroxide solution within 6min for 0.1ml of serum/urine samples. Using 0.5ml of 65% nitric acid solution the best recovery (95.3%) was obtained when 7min of effective digestion time was used. Freeze-thaw stability and long-term stability were checked. After 24 weeks 14.7% loss of iodine in urine, and 10.9% in serum samples occurred. For urine samples, better correlation (R(2)=0.9891) of various sample preparation procedures (alkaline digestion and application of OnGuard RP cartidges) was obtained. Significantly lower iodide content was found in samples taken from obese children. Serum iodine content in obese children was markedly variable in comparison with the healthy group, whereas the difference was less evident when urine samples were analyzed. The mean content in serum was 59.12±8.86µg/L, and in urine 98.26±25.93 for obese children when samples were prepared by the use of optimized alkaline digestion reinforced by microwaves. In healthy children the mean content in serum was 82.58±6.01µg/L, and in urine 145.76±31.44µg/L.