Daily dietary selenium intake of selected Brazilian population groups

Due to its essential characteristics, the daily dietary selenium intake of individuals should be monitored accurately. In the current work, daily selenium intake of different Brazilian population groups based on duplicate portion diet analysis was evaluated and compared with the new estimated average requirement values (EAR), to assess if selenium deficiency or excess could be observed in these groups. Selenium content was determined by neutron activation analysis (NAA). The average daily dietary selenium intake found was 26.3 (±8.3) ·g/day for children from the city of São Paulo, 37.4 (±16.0) ·g/day for children from Belém, 107 (±107) ·g/day for children from Macapá, 28.4 (±7.5) ·g/day for institutionalized elderly, 32 (±6) ·g/day for non-institutionalized elderly and 37 (±17) ·g/day for university students from São Paulo. Most daily dietary selenium intake range observed were below the EAR values. The values obtained for children groups from Belém and Macapá cities, whose intake levels were much higher than the recommendation, were an exception.     

Selenium and vitamin E concentrations in human milk and formula milk from Hungary

The metabolic roles of vitamin E and selenium are closely related, and to a very great extent, each can compensate for the deficiency of the other. The aim of the study was to determine and compare the Se and vitamin E (α- and γ-tocopherol) contents of breast milk and commercially available infant formulas in Hungary. The Se content was measured by instrumental neutron activation analysis (INAA), while the α-, and γ-tocopherol concentrations were determined by high performance liquid chromatography (HPLC). The mean Se concentration was 17.4±2.8 μg/L in transitional and 13.8±2.3 μg/L in mature milk. It was found that, all of the starter (ST), the follow-on (FO) and the specialized formulas (SF) had lower Se content than breast milk. Transitional breast milk resulted in a higher Se intake (14 μg/day) than mature milk (11 μg/day). The daily Se intakes in Hungarian infants were within the Recommended Dietary Allowance (RDA) range. The natural vitamin E contents of human milk were similar during the early and late lactation. Mature breast milk had 3.30±1.13 mg/L α-TE concentration and this was significantly higher than that of in ST (1.98±1.57), and FO (1.77±0.78), or in SF ready to feed preparations (1.03±0.74). The present study suggests that the formulas for the optimal development of young infants, should contain concentrations of these antioxidants on a level which is comparable to that of the human milk.     

Appraisal of venomous metals in selected crops and vegetables from industrial areas of the Punjab Province

Due to the inadequate water sources, usually sewerage water and industrial effluents are being use for irrigation of the agricultural land around the industrial areas in Pakistan wherein crops and vegetables are cultivated. As untreated effluents contain heavy elements, toxic metals and organic pollutants that may find its way through food chain to general public and may cause health hazards. It is, therefore, mandatory to assess the toxic metals in such crops and vegetables. In this regard, samples of corn, millet, cabbage, spinach and potato were collected within the vicinity of industrial areas of the Faisalabad and Gujranwala regions. The food samples were analyzed using neutron activation analysis (NAA) technique. The highest concentration values of Arsenic (1.9 ± 0.1 μg/g) and Cobalt (0.85 ± 0.01 μg/g) were found in cabbage whereas Manganese (91.6 ± 0.2 μg/g), Antimony (0.15 ± 0.03 μg/g) and Selenium (1.1 ± 0.1 μg/g) were observed in spinach and Chromium (9.63 ± 1.3 μg/g) was found in millet crop. The observed concentrations of all the toxic and heavy metals in crops and vegetables are higher than those reported in the literature.     

Selenium and mercury determination in biological samples using gamma–gamma coincidence and Compton suppression

Biological materials containing trace amounts of mercury and selenium were examined using neutron activation analysis. They were analyzed using Compton suppression and γ–γ coincidence counting. The 279 keV photopeak of activated mercury (²⁰³Hg) was analyzed in order to observe the mercury content in these samples. Selenium, an element found in many biological samples, interferes with the analysis of ²⁰³Hg when activated (⁷⁵Se). Because the selenium interference comes from a cascading emission, Compton suppression was utilized to reduce this interference. In order to fully characterize the selenium content in the samples, γ–γ coincidence was used which reduced the background and eliminated bremsstrahlung interference produced from neutron activated phosphorous through the ³¹P(n, γ)³²P reaction which is a pure beta emitter. As a result, we determined the mercury and selenium concentrations in three standard reference materials, which contain varying ratios of mercury to selenium concentrations. This study also showed that these types of concentrations can be determined from small (<500 mg) sample masses. Further work needs to be done on wet samples that require dehydration, as mercury can be lost through this process.     

Selenomethionine content of candidate reference materials

Selenium has been identified as an antioxidant of importance in the diet. Accurate determination of its chemical forms depends on the availability of suitable reference materials (RMs). Two candidate reference materials for determination of selenomethionine (Semet) in food-related materials, a standard wheat gluten sample (NIST RM 8418 Wheat Gluten) and a commercial selenium enriched yeast, have been examined by use of a gas chromatography–isotope dilution mass spectrometry (IDMS) procedure, after treatment of the matrix with 0.1 mol L^–1 hydrochloric acid containing stannous chloride, addition of CNBr, and extraction with chloroform. This procedure results in cleavage of the CH₃Se group to form volatile CH₃SeCN. Addition of isotopically enriched ⁷⁴Semet to an analytical sample enables estimation of the naturally occurring protein-bound ⁸⁰Semet by IDMS without a protein-digestion process. We found that the Wheat Gluten RM contains a significant amount of Semet as a portion of its assigned value of 2.58 μg Se_total g^–1. Commercial selenium yeast tablets are labeled as containing an elevated level of “organic selenium”, usually as Semet. The sample we investigated contained 210 μg Se_total g^–1 sample as determined separately by IDMS, measuring elemental selenium after digestion. 73% of this total (153±21 μg Se_Semet g^–1; n = 23) was present as Semet. Thus, these two materials contain significant amounts of their total selenium content as Semet and would be good candidates for further study and characterization as reference materials for determining this important food component. The CNBr reaction used will also enable the determination of Se-(methyl)selenocysteine, the biological role of which is of recent interest. In addition to matrix RMs for Semet, it is important to have standard materials of the pure substance. We have examined a sample of a candidate standard material of selenomethionine being prepared by the USP. It was confirmed that this material is pure selenomethionine. Received: 13 December 2000 / Revised: 5 March 2001 / Accepted: 12 March 2001     

Estimation of dietary iodine and bromine intakes of Ukrainians

Total diet samples for Ukrainians and Japanese were collected by duplicate portion studies in relation to the Chernobyl accident. Stable iodine and bromine in the diet samples were determined by using ICP-MS after chemical separation. Daily intakes of iodine for Ukrainians and Japanese were found to be 110±104 and 1770±1610 μg per person per day, respectively. Daily intakes of Br were as follows: Ukrainians 3.47±2.12 mg; Japanese 11.4±2.4 mg. Daily iodine intake in Ukrainians was lower than the recommended dietary allowance (150 μg) and showed the relationship of high prevalence of goiter in this area.     

Determination of selenium status using the nail biologic monitor in a canine model

Toenails and fingernails are routinely used to estimate selenium status in epidemiological studies; however, literature validating nail selenium concentration as a surrogate for critical organs is limited. In this study diets of intact male dogs were selenium supplemented at two physiological levels (3 and 6 μg/kg/day) in two different forms, selenomethionine and selenium-enriched bioformed yeast. The selenium-adequate basal diet consumed by the treatment and control groups during the 4-week run-in period and throughout the trial contained 0.3 ppm selenium. After 7 months the dogs in the two treatment groups and the control group were euthanized. Representative tissue samples from prostate, brain, liver, heart and skeletal muscle were collected, rinsed and frozen. Toenail clippings from multiple toes were also collected. Selenium was determined by neutron activation analysis using Se77m (half life = 17.4 s) at the University of Missouri Research Reactor Center. NIST SRM 1577, Bovine Liver was analyzed as a quality control. The analysts were blinded to control and treatment group assignments. As expected, tissue selenium levels increased proportionally with supplementation. A slightly greater increase in tissue selenium was observed for the purified selenomethionine compared to the bioformed yeast; however this trend was significant only for brain tissue. Toenail selenium concentrations and tissue selenium were highly correlated (p < 0.003) with Pearson coefficients of 0.759 (skeletal muscle), 0.745 (heart), 0.729 (brain), 0.723 (prostate), and 0.632 (liver). The toenail biologic monitor accurately assesses selenium status in skeletal muscle, heart, brain, prostate, and liver in the canine model.     

Determination of cyanide in blood by electrospray ionization tandem mass spectrometry after direct injection of dicyanogold

An electrospray ionization tandem mass spectrometric (ESI-MS-MS) method has been developed for the determination of cyanide (CN^–) in blood. Five microliters of blood was hemolyzed with 50 μL of water, then 5 μL of 1 M tetramethylammonium hydroxide solution was added to raise the pH of the hemolysate and to liberate CN^– from methemoglobin. CN^– was then reacted with NaAuCl₄ to produce dicyanogold, Au(CN)₂^–, that was extracted with 75 μL of methyl isobutyl ketone. Ten microliters of the extract was injected directly into an ESI-MS-MS instrument and quantification of CN^– was performed by selected reaction monitoring of the product ion CN^– at m/z 26, derived from the precursor ion Au(CN)₂^– at m/z 249. CN^– could be measured in the quantification range of 2.60 to 260 μg/L with the limit of detection at 0.56 μg/L in blood. This method was applied to the analysis of clinical samples and the concentrations of CN^– in the blood were as follows: 7.13 ± 2.41 μg/L for six healthy non-smokers, 3.08 ± 1.12 μg/L for six CO gas victims, 730 ± 867 μg for 21 house fire victims, and 3,030 ± 97 μg/L for a victim who ingested NaCN. The increase of CN^– in the blood of a victim who ingested NaN₃ was confirmed using MS-MS for the first time, and the concentrations of CN^– in the blood, gastric content and urine were 78.5 ± 5.5, 11.8 ± 0.5, and 11.4 ± 0.8 μg/L, respectively.     

Investigation of the appearance of supplemental enriched Se-76 using the human nail as a dietary monitor

The principal objective of this study was to determine if the use of a stable enriched tracer of Se-76 could be used to determine the delay time between a dietary intake of selenium and its appearance in fingernails and toenails. Selenium is an essential trace element in human nutrition. It has been studied at the Missouri University Research Reactor (MURR) for the past 15 years using an Instrumental Neutron Activation Analysis (INAA) technique. The principal route of human exposure to selenium is through the diet. Selenium concentrations of nails, blood, hair, and urine have been used as indicators of dietary selenium intake. In this study, a cohort consisting of seven men and five women ingested three selenium supplements of 150 g each over a three day period. The selenium was enriched in Se-76 (96.48%) and ingested as selenite in orange juice following an overnight fast. Fingernails and toenails were collected prior to the selenium supplementation and for several months afterward to be used as biochemical indicators. The peak⁷⁶Se concentration in the fingernails and toenails occurred at 19–23 and 16–32 weeks after supplementation, respectively.     

Fully automatic flow method for the determination of scavenging capacity against nitric oxide radicals

In the present work, a fluorimetric automatic method based on multisyringe flow injection analysis (MSFIA) was developed for in vitro evaluation of scavenging capacity against nitric oxide (NO) using 4,5-diaminofluorescein (DAF-2) as an NO-selective fluorogenic probe. The MSFIA manifold was assembled to perform the in-line generation of NO and the competitive reaction of putative scavenger molecules and DAF-2 with NO at conditions close to those found in vivo regarding temperature (37°C), pH (7.4), and concentration of NO (less than 1 μM). This approach allowed the evaluation of scavenging capacity against NO by endogenous antioxidant molecules, pharmaceutical compounds, and human plasma. IC₅₀ values were calculated for rutin (1.30 ± 0.02 μM, positive control), cysteine (321 ± 8 μM), reduced glutathione (1106 ± 93 μM), uric acid (134 ± 12 μM), dipyrone (1.36 ± 0.06 μM), and captopril (363 ± 28 μM). A high degree of automation was attained as the successive dilution of antioxidant standard solutions required for IC₅₀ assessment was performed automatically, in a dilution chamber placed in the flow system. A determination throughput of 16 h^-1 and a good precision were attained (relative standard deviation between 1.6 and 9.0%), fostering the application of the proposed method to routine/screening analysis of scavenging capacity against NO.     

Determination of enzyme activity inhibition by FTIR spectroscopy on the example of fructose bisphosphatase

A mid-infrared enzymatic assay for label-free monitoring of the enzymatic reaction of fructose-1,6-bisphosphatase with fructose 1,6-bisphosphate has been proposed. The whole procedure was done in an automated way operating in the stopped flow mode by incorporating a temperature-controlled flow cell in a sequential injection manifold. Fourier transform infrared difference spectra were evaluated for kinetic parameters, like the Michaelis–Menten constant (K _M) of the enzyme and V _max of the reaction. The obtained K _M of the reaction was 14 ± 3 g L⁻¹ (41 μM). Furthermore, inhibition by adenosine 5′-monophosphate (AMP) was evaluated, and the K _M^App value was determined to be 12 ± 2 g L⁻¹ (35 μM) for 7.5 and 15 μM AMP, respectively, with V _max decreasing from 0.1 ± 0.03 to 0.05 ± 0.01 g L⁻¹ min⁻¹. Therefore, AMP exerted a non-competitive inhibition.     

Assessment of annual effective dose from natural radioactivity intake through wheat grain produced in Faisalabad,Pakistan

Wheat is staple food of the people of Pakistan. Phosphate fertilizers, used to increase the yield of wheat, enhance the natural radioactivity in the agricultural fields from where radionuclides are transferred to wheat grain. A study was, therefore, carried out to investigate the uptake of radioactivity by wheat grain and to determine radiation doses received by human beings from the intake of foodstuffs made of wheat grain. Wheat was grown in a highly fertilized agricultural research farm at the Nuclear Institute of Agriculture and Biology (NIAB), Faisalabad, Pakistan. The activity concentration of ⁴⁰K, ²²⁶Ra and ²³²Th was measured in soil, single superphosphate (SSP) fertilizer, and wheat grain using an HPGe-based gamma-ray spectrometer. Soil to wheat grain transfer factors determined for ⁴⁰K, ²²⁶Ra and ²³²Th were 0.118 ± 0.021, 0.022 ± 0.004 and 0.036 ± 0.007, respectively, and the annual effective dose received by an adult person from the intake of wheat products was estimated to be 217 μSv.     

Separation and determination of selenium in water samples by the combination of APDC coprecipitation: X-ray fluorescence spectrometry

A sensitive and non chromatographic analytical procedure for the separation of inorganic selenium species in natural water has been performed. A combination of APDC coprecipitation and determination by an absolute thin layer Energy dispersive X-ray fluorescence spectrometry method was used. The influence of various analytical parameters such as element concentration, oxidation states and pH on the recoveries of Se (IV) was examined. The presence of organic matter and bicarbonate anions, typical components in Cuban groundwater samples, was also tested. Negligible matrix effects were observed. At pH 4 a 100% recovery was found for Se (IV). The coprecipitation recovery of the oxidized selenium species (Se (VI)) was null for the selected concentration range (5–100 μg L⁻¹). When the Se (VI) was reduced by heating the solution with 4 mol L⁻¹ HCl, quantitative recovery was also obtained. The determination of total selenium was conducted by the application of the oxidation–reduction process and the analytical procedure for Se (IV). Se (VI) content was calculated as the difference between total selenium and Se (IV). The detection limit was 0.13 μg L⁻¹. The relative standard deviation was lower than 3.5% for 5 μg L⁻¹ of Se (IV). The trueness of the method was verified by using standardized hydride generation-atomic absorption spectrometry technique. The results obtained using the EDXRF technique were in good agreement with the ones determined by HG-AAS. The proposed method was applied to the determination of Se (IV) in surface water and groundwater samples.     

Kinetics of cordierite crystallization from diphasic gels

Diphasic cordierite gels were prepared from colloidal silica, aluminum and magnesium nitrates and citric acid. The mechanism of xerogel decomposition was studied by infrared spectroscopy (FT-IR) and thermal gravimetric analysis (TGA). The thermal decomposition of the xerogel forms a solid mixture of MgO, Al₂O₃ and SiO₂ at around 250 °C. Cordierite crystallization was studied by X-ray diffraction (XRD) and differential thermal analysis (DTA). Xerogels were initially thermally treated, and this sample crystallized to μ-cordierite at 850 °C, at 900 °C α-cordierite crystallizes and at 1150 °C α-cordierite is the major phase and μ-cordierite is totally consumed. The apparent activation energy for cordierite crystallization process was determined based on the Johnson-Mehl-Avrami-Kolmogorov (JMAK) theory, Ligero methods and the Arrhenius law for dependence of activation energy with temperature. The apparent activation energy was (466.8 ± 34.3) kJ/mol, the exponent of Avrami was (1.9 ± 0.2) and the frequency factor was (1.55 × 10²⁰) s⁻¹. The Avrami value indicates a nucleation controlled process, which can be a consequence of the high xerogel homogeneity, a consequence of the early and simultaneous formation of the MgO, Al₂O₃ and SiO₂ mixture.     

Analysis of mono-phosphate nucleotides as a potential method for quantification of DNA using high performance liquid chromatography–inductively coupled plasma-mass spectrometry

The determination of total deoxyribonucleic acid (DNA) concentration is of great importance in many biological and bio-medical analyses. The quantification of DNA is traditionally performed by UV spectroscopy; however the results can be affected greatly by the sample matrix. The proposed method quantifies phosphorus in digested calf thymus DNA and human DNA by high performance liquid chromatography (HPLC) combined with inductively coupled plasma mass spectrometry (ICP-MS). The method presented showed excellent baseline separation between all four DNA mono-nucleotides and 5′UMP. The ability of LC-ICP-MS to provide an internal check that only DNA derived phosphorus was counted in the assay was demonstrated by establishing a mass balance between the total phosphorous signal from undigested DNA and that from the speciated DNA. Column recoveries ranging from 95% to 99% for phosphorus resulted in a mass balance of 95% ± 0.5% for standard nucleotides, determined by LC-ICP-MS, compared to total DNA determined by flow injection coupled to ICP-MS (FI-ICP-MS). The method for quantification was validated by analysis of NIST SRM 2,372; a total speciated DNA recovery of 52.1 ng/μL, compared with an expected value of 53.6 ng/μL, was determined by external calibration. From repeat measurements, a mass balance of 97% ± 0.5% for NIST DNA was achieved. The method limits of detection for individual nucleotides were determined between 0.8 and 1.7 μg L⁻¹ (³¹P) for individual nucleotides by LC-ICP-MS, and 360 ng L⁻¹ for 5′AMP by direct nebulisation.     

Preparation of antibodies and development of a sensitive immunoassay with fluorescence detection for triazine herbicides

Specific polyclonal antibodies against s-triazine herbicides were obtained by preparing immunogens coupling home-synthesized haptens derivatives of simazine (6-chloro-N-ethyl-N′-ethyl-1,3,5-triazine-2,4-diamine) to lysine groups of hemocyanin from keyhole limpets and bovine serum albumin carrier proteins. Three highly sensitive rabbit antisera were obtained and evaluated with a battery of six enzyme tracers derived from triazine structures in an optimized ELISA format. The antiserum As8 and the HRP-2f tracer, which yield the best assay sensitivity for simazine (detection limit 0.11 ± 0.02 μg L⁻¹, IC₅₀ 0.88 ± 0.04 μg L⁻¹), were applied to the development of a sensitive flow-through immunoassay for the analysis of this herbicide. The automated assay was based on a direct competitive immunosorbent assay and fluorescence detection. The optimized method presents an IC₅₀ value of 0.35 ± 0.04 μg L⁻¹ with a detection limit of 1.3 ± 0.9 ng L⁻¹ and a dynamic range from 0.010 to 7.5 μg L⁻¹ simazine. The generic nature of the antiserum was shown by good relative cross-reactivities with other triazines such as atrazine (420%) or propazine (130%) and a lower response to terbutylazine (6.4%) and desethyl-atrazine (2.2%). No cross-reactivity was obtained for nonrelated pesticides such as 2,4-dichlorophenoxyacetic acid or linuron and the assay could be applied as a screening method for triazine herbicides. The total analysis time was 30 min per determination and the immunosensor could be reused for more than 150 cycles without significant loss of activity. The immunosensor has been successfully applied to the direct analysis of simazine in surface water samples at the nanogram per liter level. The results obtained by comparative analysis of the immunosensor with a chromatographic procedure for triazines showed a close correspondence.     

Selenium determination in tissue samples of Nile tilapia using ultrasound-assisted extraction

This paper proposes a method to determine selenium in samples of fish muscle and liver tissue using ultrasound assisted extraction process, and analysed by graphite furnace atomic absorption spectrometry (GFAAS). The selenium content was extracted by 0.10 M HCl at the optimal extraction conditions which were established as follows: sample mass of 100 mg; granulometry of the sample <60 μm; sonication time of five 40 s cycles; and sonication power of 136 W. The selenium determinations were performed by GFAAS, at a drying temperature of 120°C/250°C, pyrolysis temperature of 1300°C, atomization temperature of 2300°C, and cleaning temperature of 2800°C. Palladium nitrate was used as a chemical modifier coinjected with the samples, and tungsten as a permanent modifier. The concentration of selenium determined in the pool of fish muscle and liver tissue were 280.4±4.2 e 592.3±6.7 μg kg⁻¹, respectively. The accuracy and precision of the proposed extraction method were evaluated using certified standard Bovine Muscle — NIST 8414. The results obtained by the ultrasonic extraction method were equivalent to those obtained by the method of acid mineralization of samples in a microwave oven     

Speciation of four selenium compounds using high performance liquid chromatography with on-line detection by inductively coupled plasma mass spectrometry or flame atomic absorption spectrometry

An analytical method for the speciation of selenomethionine, selenocystine, selenite and selenate by high performance liquid chromatography (HPLC) with atomic spectrometric detection is presented. An organic polymeric strong anion exchange column was used as the stationary phase in combination with an aqueous solution of 6 mmol L^–1 of salicylate ion at pH 8.5 as the mobile phase which allowed the isocratic separation of the four selenium analytes within 8 minutes. The separated selenium species were detected on-line by flame atomic absorption spectrometry (FAAS) or inductively coupled plasma mass spectrometry (ICP-MS). The signal-to-noise ratio of the FAAS detector was optimized using a hydrogen-argon entrained-air flame and a slotted-tube atom trap (STAT) in the flame. The limit of detection (3 σ) achieved by the HPLC-FAAS system was 1 mg L^–1 of selenium (100 μL injections) for each of the four selenium species. More powerful selenium detection was achieved using an ELAN 5000 ICP-MS instrument. Selenium was measured at m/z = 82. The ICP-MS signal intensity was enhanced by a factor of 3–4 after addition of 3% methanol to the chromatographic mobile phase and by using an increased plasma power input of 1300 W. The limit of detection achieved under these conditions was 1 μg L^–1 (100 μL injections). The HPLC-ICP-MS system was used for selenium speciation of selenite and selenate in aqueous solutions during a BCR certification exercise and for selenium speciation in the certified reference material, BCR No. 402 White Clover. Extraction experiments revealed that the selenium species in the biological material were extractable only in the presence of water in the extraction medium. The results indicated that selenate and a compound of unknown identity U were present in the plant sample. Received: 4 September 1996 / Accepted: 12 December 1996     

Simultaneous ion chromatographic determination of selenium and metal ions in waters at trace level

Summary An ion-chromatographic procedure is described for the determination of selenium (VI) at μg L⁻¹ level in the presence of anions and heavy metal ions. Maximum permissible concentrations and effects from each interfering substance were investigated for the Se concentration range 12.5–1,000 μg L⁻¹. The method, optimized for the detection of SeO ₄ ²⁻ , gives results suitable for speciation analysis. Total selenium can be determined after complete conversion to selenate ion by oxidation with KMnO₄. The detection limit of selenium is 4.8 μg L⁻¹ (0.96 ng for 200 μL sample). Paper presented at the 41st Pittsburgh Conference, New York, March 5–9, 1990.     

Ultra-fast two-dimensional microchip electrophoresis using SDS μ-CGE and microemulsion electrokinetic chromatography for protein separations

A poly(methyl methacrylate) microfluidic chip was used to perform a two-dimensional (2-D) separation of a complex protein mixture in short development times. The separation was performed by combining sodium dodecyl sulfate micro-capillary gel electrophoresis (SDS μ-CGE) with microemulsion electrokinetic chromatography (μ-MEEKC), which were used for the first and second dimensions, respectively. Fluorescently labeled Escherichia coli cytosolic proteins were profiled by this 2-D approach with the results compared to a similar 2-D separation using SDS μ-CGE × μ-MEKC (micelle electrokinetic chromatography). The relatively short column lengths (effective length = 10 mm) for both dimensions were used to achieve separations requiring only 220 s of development time. High spot production rates (131 ± 11 spots min⁻¹) and reasonable peak capacities (481 ± 18) were generated despite the fact that short columns were used. In addition, the use of μ-MEEKC in the second dimension was found to produce higher peak capacities compared to μ-MEKC (481 ± 18 for μ-MEEKC and 332 ± 17 for μ-MEKC) due to the higher plate numbers associated with μ-MEEKC.