Katarzyna Wrobel

Capillary electrophoresis-inductively coupled plasma-mass spectrometry: an attractive complementary technique for elemental speciation analysis.

Some basic and practical aspects of interfacing capillary electrophoresis to inductively coupled plasma-mass spectrometry (CE-ICP-MS) are reviewed in this article with emphasis on the use of this hyphenated technique for elemental speciation analysis. The principles behind the techniques of both CE and ICP-MS are introduced. The interfacing of CE to ICP-MS is discussed including several devices and nebulizers reported in literature. A brief account of their advantages and limitations is given. The various CE-ICP-MS applications for elemental speciation analysis are also reviewed. Some issues concerning the future of CE-ICP-MS for the elemental speciation analyses are discussed.

Dietary Advanced Glycation End Products and Their Role in Health and Disease

Over the past 2 decades there has been increasing evidence supporting an important contribution from food-derived advanced glycation end products (AGEs) to the body pool of AGEs and therefore increased oxidative stress and inflammation, processes that play a major role in the causation of chronic diseases. A 3-d symposium (1st Latin American Symposium of AGEs) to discuss this subject took place in Guanajuato, Mexico, on 1-3 October 2014 with the participation of researchers from several countries. This review is a summary of the different presentations and subjects discussed, and it is divided into 4 sections. The first section deals with current general knowledge about AGEs. The second section dwells on mechanisms of action of AGEs, with special emphasis on the receptor for advanced glycation end products and the potential role of AGEs in neurodegenerative diseases. The third section discusses different approaches to decrease the AGE burden. The last section discusses current methodologic problems with measurement of AGEs in different samples. The subject under discussion is complex and extensive and cannot be completely covered in a short review. Therefore, some areas of interest have been left out because of space. However, we hope this review illustrates currently known facts about dietary AGEs as well as pointing out areas that require further research.

Determination of total aluminum, chromium, copper, iron, manganese, and nickel and their fractions leached to the infusions of black tea, green tea, Hibiscus sabdariffa, and Ilex paraguariensis (mate) by ETA-AAS.

Total aluminum, chromium, copper, iron, manganese, and nickel were determined in black tea, green tea, Hibiscus sabdariffa, and Ilex paraguariensis (mate) by electrothermal atomic absorption spectrometry after nitric/perchloric acid digestion. In each case, one ground sample of commercially available leafy material was prepared and three 0.5-g subsamples were run in parallel. The infusions were also analyzed and the percentage of each element leached into the liquor was evaluated. The obtained results indicated that hibiscus and mate contained lower levels of aluminum (272±19 µg/g and 369±22 µg/g, respectively) as referred to black tea (759±31 µg/g) or green tea (919±29 µg/g) and suggested that mate drinking could be a good dietary source of essential micronutrient manganese (total content 2223±110 µg/g, 48.1% leached to the infusion). It was also found that the infusion of hibiscus could supply greater amounts of iron (111±5 µg/g total, 40.5% leached) and copper (5.9±0.3 µg/g total, 93.4% leached) as compared to other infusions. Moreover, it was found that the percentage of element leached to the infusion was strongly related to the tannins content in the beverage (correlation coefficients >0.82 with the exception for nickel); for lower tannins level, better leaching was observed.

Aluminium and silicon speciation in human serum by ion-exchange high-performance liquid chromatography–electrothermal atomic absorption spectrometry and gel electrophoresis

Speciation of aluminium and silicon in serum was studied by a reliable and sensitive high-performance liquid chromatographic-electrothermal atomic absorption spectrometric (HPLC-ETAAS) hybrid method, based on the use of a polymeric anion-exchange column (Protein-Pak DEAE-5PW). This polymer-based column minimizes the risk of aluminium losses and of silicon contamination from the column during separation. The results obtained were compared with the results of previous studies carried out using different, complementary techniques including ultramicrofiltration, gel filtration and silica-based column for HPLC. In order to ascertain which protein(s) of serum actually bind(s) aluminium, gel electrophoresis was employed for the further separation of the column fractions obtained by HPLC and aluminium was determined in separate aliquots of the same fractions. From all the experiments, it appears that transferrin (Tf) is the only serum protein that binds aluminium and it contains about 90% of total serum aluminium. It was also confirmed that in the presence of desferrioxamine (DFO). aluminium is partly displaced from its complex with transferrin to a low molecular mass AL-DFO complex. Aluminum citrate seems to be the main low molecular mass aluminium species in serum, amounting to about (12 +/- 5% of the total aluminium in an aluminium-loaded serum sample. The proposed speciation procedure permits the simultaneous identification and determination of three aluminium species in metal-spiked serum (Al-Tf, Al-DFO and AI-citrate). The result for silicon suggest that it seems to be unspecifically adsorbed to several serum proteins and its speciation is not affected by the presence of DFO.(ABSTRACT TRUNCATED AT 250 WORDS)

Trace elements status in diabetes mellitus type 2: Possible role of the interaction between molybdenum and copper in the progress of typical complications

It is well established that both, the deficiency and possible overload of mineral micronutrients have adverse health effects. It is also generally accepted that non-essential xenobiotics contribute to oxidative damage, which is considered one of the principal factors in diabetes and its complications. The purpose of this work was to gain an insight on the global role of metal/metalloids in the progress of diabetes mellitus type 2. In such approach, aluminum, vanadium, chromium, manganese, cobalt, nickel, copper, zinc, arsenic, selenium, molybdenum, mercury, cadmium and lead were determined by inductively coupled plasma-mass spectrometry (ICP-MS) in serum and urine of 76 diabetic patients (age 52 ± 8 years, 5-16 years of DM2, 52 subjects with slight-to-moderate complications and 24 with severe complications). A series of anthropometric and clinical parameters usually evaluated in the follow-up of patients were assessed by standard methods. Statistical analysis (unpaired t-test, analysis of correlation and principal component analysis) was then carried out in search of possible relationships existing among metals/metalloids and these parameters. The results obtained suggest that antagonistic interaction between molybdenum and copper might be involved in the progress of diabetes complications.

Serum selenium and glutathione peroxidase concentrations in type 2 diabetes mellitus patients

AIMS Antioxidant selenium (Se) properties and, its protective role against oxidative damage play an important role in diabetic complications. Our objective was to gain further insight on a link between selenium status and diabetic nephropathy. METHODS We assessed glutathione peroxidase (GPx) and Se in type 2 diabetes mellitus patients with microalbuminuria (MA) (group 1), without microalbuminuria (group 2), and in control subjects (group 3). Glucose, urea, creatinine and glycated hemoglobin tests were tested in sera. A complete clinical record was elaborated. RESULTS For diabetic patients both, the time from diagnosis and plasma glucose concentration were higher in group 1 as compared to group 2. Control group showed higher serum Se concentrations as compared to the diabetic groups. The two groups of diabetic patients showed similar serum Se levels. Serum concentration of GPx was significantly lower in group 1 as compared to groups 2 and 3. Microalbuminuria (MA) test showed a positive correlation with glucose, and a negative relationship with serum Se and GPx. Multiple regression revealed an inverse relationship between selenium or GPx in serum and the results of the MA test. CONCLUSIONS Our results suggest that lower Se and GPx levels in diabetic patients may be implicated in the diabetic nephropathy.

Studying the distribution pattern of selenium in nut proteins with information obtained from SEC-UV-ICP-MS and CE-ICP-MS

In this work, size exclusion chromatography (SEC) with UV and inductively coupled plasma mass spectrometry (ICP-MS) detection was used to study the association of selenium to proteins present in Brazil nuts (Bertholletia excelsa) under five different extraction conditions. As expected, better solubilization of proteins was observed using 0.05molL(-1) sodium hydroxide and 1% sodium dodecylsulfate (SDS) in Tris/HCl buffer (0.05molL(-1), pH 8) as compared to 0.05molL(-1) HCl, 0.05molL(-1) Tris/HCl or hot water (60 degrees C). Due to non-destructive character of Tris-SDS treatment, this was applied for studying molecular weight (MW) distribution patterns of selenium-containing nut proteins. Three different SEC columns were used for obtaining complete MW distribution of selenium: Superdex 75, Superdex Peptide, and Superdex 200 were tested with 50mmolL(-1) Tris buffer (pH 8), 150mmolL(-1) ammonium bicarbonate buffer (pH 7.8), phosphate (pH 7.5), and CAPS (pH 10.0) mobile phases. Using Superdex 200 column, the elution of at least three MW fractions was observed with UV detection (200-10kDa) and ICP-MS chromatogram showed the co-elution of selenium with the two earlier fractions. The apparent MWs of these selenium-containing fractions were respectively about 107 and 50kDa, as evaluated from the column calibration. For further characterization of individual selenium species, the defatted nuts were hydrolyzed with proteinase K and analyzed by capillary electrophoresis (CE) with ICP-MS detection. The suitability of CE for the separation of selenite, selenate, selenocystine and selenomethionine in the presence of the nut sample matrix is demonstrated. Complete separation of the above mentioned selenium species was obtained within a migration time of 7min. In the analysis of nut extracts with CE-ICP-MS, selenium was found to be present mainly as selenomethionine.

Selenium speciation in low molecular weight fraction of Se-enriched yeasts by HPLC-ICP-MS: detection of selenoadenosylmethionine

In this study, selenium speciation in low molecular weight (LMW) fraction of Se-enriched yeasts was investigated assuming the presence of selenoadenosylmethionine (AdoSeMet). In order to minimize the risk of species decomposition, the pretreatment procedure consisted of homogenization of fresh cells and precipitation of proteins with perchloric acid at 0 °C. The species separation was achieved by ion pairing HPLC (hexanesulfonic acid) with UV (254 nm) and ICP-MS detection. The assignment of chromatographic peaks was by spiking the sample with both the commercial and synthesized standards. The synthesis of AdoSeMet and selenoadenosylhomocysteine (AdoSeHcy) was performed and their structures confirmed by ES-MS analysis. These two selenium species were identified in yeasts extract and the presence of Se-cystine was also observed. Significantly lower ratio between AdoSeMet and AdoSeHcy was observed in yeast extracts as compared to the ratio between sulfur analogs. This observation could contribute to further understanding of the selenium role in biomethylation processes.

Dietary advanced glycation end products restriction diminishes inflammation markers and oxidative stress in patients with type 2 diabetes mellitus.

The augmented consumption of dietary advanced glycation end products (dAGEs) has been associated with increased oxidative stress and inflammation, however, there is insufficient information over the effect on insulin resistance. The objective of the present study is to investigate the effect of dAGEs restriction on tumor necrosis factor-α (TNF-α), malondialdehyde, C-reactive protein (CRP), and insulin resistance in DM2 patients. We carried out a randomized 6 weeks prospective study in two groups of patients: subjects with a standard diet (n = 13), vs low dAGEs (n = 13). At the beginning and the end of study, we collected anthropometric measurements, and values of circulating glucose, HbA1c, lipids, insulin, serum AGEs, CRP, TNF-α and malondialdehyde. Anthropometric measurements, glucose, and lipids were similar in both groups at base line and at the end of the study. Estimation of basal dAGEs was similar in both groups; after 6 weeks it was unchanged in the standard group but in the low dAGEs group decreased by 44% (p<0.0002). Changes in TNF-α levels were different under standard diet (12.5 ± 14.7) as compared with low dAGEs (−18.36 ± 17.1, p<0.00001); changes in malondialdehyde were different in the respective groups (2.0 ± 2.61 and −0.83 ± 2.0, p<0.005) no changes were found for insulin levels or HOMA-IR. In conclusion, The dAGEs restriction decreased significantly TNF-α and malondialdehyde levels.

Analytical speciation of mercury in fish tissues by reversed phase liquid chromatography-inductively coupled plasma mass spectrometry with Bi3+ as internal standard.

In this work, the quantification of two mercury species (Hg(2+) and CH(3)Hg(+)) in fish tissues has been revisited. The originality of our approach relies on the use of Bi(3+) as internal standard (IS) and on the modification of typical extraction conditions. The IS (125 microl, 1000 microg l(-1) Bi(3+)) was added to the aliquot of fresh fish tissue (400-500 mg). A high-speed blender and ultrasound-assisted homogenization/extraction was carried out in the presence of perchloric acid (1.5 ml, 0.6 mol l(-1)), l-cysteine (500 microl, 0.75 mol l(-1)) and 500 microl toluene:methanol (1:1). Perchloric acid was used for protein denaturation and precipitation, toluene helped to destroy lipid structures potentially sequestering CH(3)Hg(+), L-cysteine was used to form water-soluble complexes with Bi(3+), Hg(2+) and CH(3)Hg(+). The excess of perchloric acid was eliminated by addition of potassium hydroxide (pH 5 with acetic acid). The obtained extract, was diluted with the mobile phase (1:1) and introduced (20 microl) to the reversed phase HPLC-ICP-MS system. The separation was achieved by isocratic elution (2.5 mmol l(-1) cysteine, 12.5 mmol l(-1) (NH(4))(2)HPO(4), 0.05% triethylamine, pH 7.0:methanol (96:4)) at a flow rate 0.6 ml min(-1). Column effluent was on-line introduced to ICP-MS for specific detection of (202)Hg, (200)Hg and (209)Bi. Analytical signal was defined as the ratio between (202)Hg/(209)Bi peak areas. The detection limits evaluated for Hg(2+) and CH(3)Hg(+) were 0.8 and 0.7 microg l(-1). Recovery of the procedure, calculated as the sum of species concentrations found in the sample with respect to total ICP-MS-determined Hg was 91.9% for king mackerel muscle and 89.5% for red snapper liver. In the standard addition experiments, the recovery results were 98.9% for Hg(2+) and 100.6% for CH(3)Hg(+). It should be stressed that the use of Bi(3+) as IS enabled to improve analytical performance by compensating for incomplete extraction and for imprecision of sample handling during relatively non-rigorous protocol.