Anion-exchange Chromatographic Separation Research Articles

The Efficiency Improvement of Electrodeposition of High-Purity Zn by a Proper Electrolyte Solution Preparation from Spent Pickling Liquor by Anion-Exchange Separation

A better procedure for recovering high-purity Zn from the SPL solution has been devised. To provide a suitable electrolyte for the efficient cathodic deposition of high-purity Zn, complete anion-exchange purification had been done by chromatographic technique. The process was then continued by galvanostatic electrodeposition to determine the efficiencies. As the SPL needs to be neutralized before any attempts of Zn electrodeposition, NaCl may provide the required source of Cl- ions for complex formation. The equilibrium studies of the anion-exchange distribution in such solutions showed similar tendencies to those already known in HCl media. The anion-exchange chromatographic separation was devised – with the finer tuning of the parameters – according to the equilibrium result, but other physical parameters were also examined for improving the efficiency of separation and the recovery of Zn. The devised anion-exchange separation in chloride media can purify the SPL, containing Zn as the valuable component and higher concentrations of Fe as the main impurity. The most important impurities (Fe, Mn, Ni, Al and Co) can be efficiently separated in a dissolved state. At the same time, Zn is sorbed in the strongly basic anion[1]exchange resin at a moderate Cl- ion concentration (~ 2 M). Stirring with Fe chips provides a reduction of iron to the Fe(II) state. The remaining problem is minor copper contamination in the same effluent fraction as Zn, which however can be eliminated by a simple additional step of cementation or directly by pre-electrolysis.

Liquid Anion Exchange Chromatographic Extraction and Separation of Osmium(VIII) with 4-(4-Methoxy-benzylideneimino)-5-methyl-4H-1,2,4-triazole-3-thiol in Hydrochloric Acid Medium

A novel method is proposed for the extraction of microgram level concentration of osmium(VIII) from hydrochloric acid medium with 4-(4-methoxybenzylideneimino)-5-methyl-4H- 1,2,4-triazole-3-thiol (MBIMTT) dissolved in chloroform as an extractant. The osmium(VIII) from the organic phase is stripped with mixture of thiourea and 2 M hydrochloric acid and determined spectrophotometric method. The method affords the binary separation and determination of osmium(VIII) from synthetic mixture. The method is successfully applied for the separation and determination of osmium from the alloys. The method is highly selective, simple and reproducible.

Identification of trace levels of selenomethionine and related organic selenium species in high-ionic-strength waters.

A new anion-exchange chromatographic separation method was used for the simultaneous speciation analysis of selenoamino acids and the more ubiquitous inorganic selenium oxyanions, selenite and selenate. For quantification, this separation was coupled to inductively coupled plasma-mass spectrometry to achieve an instrumental detection limit of 5 ng Se L(-1) for all species. This chromatographic method was also coupled to electrospray tandem mass spectrometry to observe the negative ion mode fragmentation of selenomethionine and one of its oxidation products. Low detection limits were achieved, which were similar to those obtained using inductively coupled plasma-mass spectrometry. An extensive preconcentration and cleanup procedure using cation-exchange solid-phase extraction was developed for the identification and quantification of trace levels of selenomethionine in environmental samples. Preconcentration factors of up to five were observed for selenomethionine, which in addition to the removal of high concentrations of sulphate and chloride from industrial process waters, allowed for an unambiguous analysis that would have been impossible otherwise. Following these methods, selenomethionine was identified at an original concentration of 3.2 ng Se L(-1) in samples of effluent collected at a coal-fired power plant's biological remediation site. It is the first time that this species has been identified in the environment, outside of a biological entity. Additionally, oxidation products of selenomethionine were identified in river water and laboratory algal culture samples. High-resolution mass spectrometry was employed to postulate the chemical structures of these species.

Determination of deoxyribonucleoside triphosphate concentrations in yeast cells by strong anion-exchange high-performance liquid chromatography coupled with ultraviolet detection.

DNA polymerase assays are commonly used for the detection of deoxyribonucleoside triphosphates (dNTPs) in biological samples. For better specificity and accuracy, high-performance liquid chromatography (HPLC) methods have been developed for the analysis of the four dNTPs in complex samples. Here we describe a simple method using isocratic strong anion-exchange (SAX) chromatographic separation coupled with ultraviolet detection (UV) for the analysis of the four dNTPs in budding yeast Saccharomyces cerevisiae. This method can be applied to other species of yeast or bacteria.

Absolute quantification of superoxide dismutase in cytosol and mitochondria of mice hepatic cells exposed to mercury by a novel metallomic approach

In the last years, the development of new methods for analyzing accurate and precise individual metalloproteins is of increasing importance, since numerous metalloproteins are excellent biomarkers of oxidative stress and diseases. In that way, methods based on the use of post column isotopic dilution analysis (IDA) or enriched protein standards are required to obtain a sufficient degree of accuracy, precision and high limits of detection. This paper reports the identification and absolute quantification of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) in cytosol and mitochondria from mice hepatic cells using a innovative column switching analytical approach. The method consisted of orthogonal chromatographic systems coupled to inductively coupling plasma-mass spectrometry equipped with a octopole reaction systems (ICP-ORS-MS) and UV detectors: size exclusion fractionation (SEC) of the cytosolic and mitochondrial extracts followed by online anion exchange chromatographic (AEC) separation of Cu/Zn containing species. After purification, Cu,Zn-SOD was identified after tryptic digestion by molecular mass spectrometry (MS). The MS/MS spectrum of a doubly charged peptide was used to obtain the sequence of the protein using the MASCOT searching engine. This optimized methodology reduces the time of analysis and avoids the use of sample preconcentration and clean-up procedures, such as cut-off centrifuged filters, solid phase extraction (SPE), precipitation procedures, off-line fractions insolates, etc. In this sense, the method is robust, reliable and fast with typical chromatographic run time less than 20min. Precision in terms of relative standard deviation (n=5) is of 3–5% and detection limits is 0.21ngCug−1.The application of the methodology to hepatic cells from mice exposed to inorganic mercury reveals decreased levels of Cu,Zn-SOD in cytosolic and mitochondrial extracts, as a consequence of the oxidative stress caused by this toxic metal. Additionally, the quantification of mitochondrial Cu,Zn-SOD in hepatic cells from Mus musculus has been carried out for the first time.

Expanding resolution of metalloprotein separations from soybean seeds using 2D-HPLC-ICP-MS and SDS-PAGE as a third dimension

This work reports on the use of a three dimensional separation system to enhance metalloprotein information when considering soybean seeds. Separations using size exclusion chromatography (SEC) allowed identification of three metal fractions. Following an anion exchange (AEX) chromatographic separation in the second dimension, the resultant sub-fractions were lyophilized and subjected to a third dimension of separation using a polyacrylamide gel electrophoresis (SDS-PAGE). After the separation, the bands were digested, and, in addition to others, the following proteins, previously associated with metals, were identified: 3-lipoxygenase A chain (soybean) complex with 13(S)-hydroperoxy-9(Z),11(E)-octadecadienoic acid, beta-amylase [Glycine max], seed lipoxygenase-1, lipoxygenase [G. max], seed lipoxygenase-2 (Pisum sativum) and beta-conglycinin. Techniques presenting high resolution are desired due to their capability in resolving great amount of signals (responses) generated from hundreds of proteins generally found in different samples. To the best of our knowledge, this is the first time that bidimensional chromatographic system which allied to another separation dimension is applied for improving protein identification, so that higher number and different proteins were found when comparing 2D dimension with 3D dimension. In fact, this strategy is welcoming in proteomics studies, in order to improve the comprehension of those systems that present large number of proteins. This article is part of a Special Issue entitled: Environmental and structural proteomics.

Rapid Multisample Analysis for Simultaneous Determination of Anthropogenic Radionuclides in Marine Environment

An automated multisample processing flow injection (FI) system was developed for simultaneous determination of technetium, neptunium, plutonium, and uranium in large volume (200 L) seawater. Ferrous hydroxide coprecipitation was used for the preliminary sample treatment providing the merit of simultaneous preconcentration of all target radionuclides. Technetium was separated from the actinides via valence control of technetium (as Tc(VII)) in a ferric hydroxide coprecipitation. A novel preseparation protocol between uranium and neptunium/plutonium fractions was developed based on the observation of nearly quantitative dissolution of uranium in 6 mol/L sodium hydroxide solution. Automated extraction (TEVA for technetium and UTEVA for uranium) and anion exchange (AGMP-1 M for plutonium and neptunium) chromatographic separations were performed for further purification of each analyte within the FI system where four samples were processed in parallel. Analytical results indicate that the proposed method is robust and straightforward, providing chemical yields of 50-70% and improved sample throughput (3-4 d/sample). Detection limits were 8 mBq/m(3) (0.013 pg/L), 0.26 μBq/m(3) (0.010 fg/L), 23 μBq/m(3) (0.010 fg/L), 84 μBq/m(3) (0.010 fg/L) and 0.6 mBq/m(3) (0.048 ng/L) for (99)Tc, (237)Np, (239)Pu, (240)Pu and (238)U for 200 L seawater, respectively. The unique feature of multiradionuclide and multisample simultaneous processing vitalizes the developed method as a powerful tool in obtaining reliable data with reduced analytical cost in both radioecology studies and nuclear emergency preparedness.

Determination of mercurial species in fish by inductively coupled plasma mass spectrometry with anion exchange chromatographic separation

This work demonstrated the feasibility of mercury speciation analysis by anion exchange chromatographic separation with inductively coupled plasma mass spectrometry detection. For the first time, by complexing with the mobile phase containing 3-mercapto-1-propanesulfonate into negatively charged complexes, fast separation of inorganic mercury (Hg2+), monomethylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg) was achieved within 5min on a 12.5-mm strong anion exchange column. The detection limits for Hg2+, MeHg, EtHg and PhHg were 0.008, 0.024, 0.029 and 0.034μgL−1, respectively. The relative standard deviations of peak height and peak area (5.0μgL−1 for each Hg species) were all below 3%. The determined contents of Hg2+, MeHg and total Hg in a certified reference material of fish tissue by the proposed method were in good accordance with the certified values with satisfactory recoveries. The relative errors for determining MeHg and total mercury were −2.4% and −1.2%, respectively, with an acceptable range for spike recoveries of 94–101%. Mercury speciation in 11 fish samples were then analyzed after the pretreated procedure. The mercury contents in all fish samples analyzed were found compliant with the criteria of the National Standards of China.

Centrifugal microfluidic platform for radiochemistry: Potentialities for the chemical analysis of nuclear spent fuels

The use of a centrifugal microfluidic platform is for the first time reported as an alternative to classical chromatographic procedures for radiochemistry. The original design of the microfluidic platform has been thought to fasten and simplify the prototyping process with the use of a circular platform integrating four rectangular microchips made of thermoplastic. The microchips, dedicated to anion-exchange chromatographic separations, integrate a localized monolithic stationary phase as well as injection and collection reservoirs. The results presented here were obtained with a simplified simulated nuclear spent fuel sample composed of non-radioactive isotopes of Europium and Uranium, in proportion usually found for uranium oxide nuclear spent fuel. While keeping the analytical results consistent with the conventional procedure (extraction yield for Europium of ≈97%), the use of the centrifugal microfluidic platform allowed to reduce the volume of liquid needed by a factor of ≈250. Thanks to their unique “easy-to-use” features, centrifugal microfluidic platforms are potential successful candidates for the downscaling of chromatographic separation of radioactive samples (automation, multiplexing, easy integration in glove-boxes environment and low cost of maintenance).

Liquid Anion Exchange Chromatographic Extraction and Separation of Platinum(IV) with n‐Octylaniline as a Metallurgical Reagent: Analysis of Real Samples

A simple and selective method was developed for the determination of platinum(IV) with n‐octylaniline in toluene. In present study, the use of n‐octylaniline in toluene for the extraction of platinum(IV) from ascorbate media was carried out. The effect of various parameters, such as pH, equilibrium time, extractant concentration, and organic solvent on the extraction has been discussed. The back extraction of platinum(IV) has been performed. On the basis of slope analysis, the composition of the extracted species was determined as [RR′NH2+ Pt(Succinate)2−](org). The interfering effects of various cations and anions were also studied, and the selectivity of the method is enhanced by using suitable masking agents. The proposed method is rapid, reproducible and successfully applied for the determination of platinum(IV) in binary and synthetic mixtures. The separation of pt(IV) from other associated metals has been studied. Comparison of the results with those obtained using an atomic absorption spectrophotometer also tested the validity of the method.

Liquid Anion Exchange Chromatographic Extraction and Separation of Platinum (IV) with N-Octylaniline as an Metallurgical Reagent: Analysis of Real Samples

A simple and selective method was developed for the determination of platinum (IV) with n-octylaniline in toluene. In present study, the use of n-octylaniline in toluene for the extraction of platinum (IV) from ascorbate media was carried out. The effect of various parameters, such as pH, equilibrium time, extractant concentration and organic solvent on the extraction has been discussed. The back extraction of has been performed. On the basis of slope analysis, the composition of the extracted species was determined as [RR’NH2 + Pt (Succinate)2 -](org). The interfering effects of various cations and anions were also studied, and use of suitable masking agents enhances the selectivity of the method. The proposed method is rapid, reproducible and successfully applied for the determination of platinum (IV) in binary and synthetic mixtures. The separation of platinum (IV) from other associated metals has been studied. Comparison of the results with those obtained using an atomic absorption spectrophotometer were also tested the validity of the method.

Fabrication of electrolytic cell for online post-column electrochemical derivatization in ion chromatography

An electrolytic cell (EC), composed of two ruthenium-plated titanium electrodes separated by cation-exchange membranes, was fabricated and evaluated for online postcolumn derivatization in ion chromatography (IC). Folic acid (FA) and methotrexate (MTX) were preliminarily used as prototype analytes to test the performance of EC. After separation by an anion exchange column, FA and MTX, which emit very weak fluorescence when excited, were electrochemically oxidized online in the anode chamber of the EC. The compounds with strong fluorescence, which are oxidation products, were detected by the fluorescence detector. The phosphate buffer solution (100mM KH2PO4) served as an optimal eluent for anion exchange chromatographic separation and a suitable supporting electrolyte for electro-oxidation, leading to ideal compatibility between IC separation and the postcolumn electrochemical derivatization. For the presently proposed method, the linear ranges were from 0.01mgL−1 to 5mgL−1 for both FA and MTX. The detection limits of FA and MTX were 1.8and 2.1μgL−1, and the relative standard deviations (RSD, n=7) were 2.9% and 3.6%, respectively. The method was applied for the simultaneous determination of FA and MTX in the plasma of patients being treated for rheumatoid arthritis. The determination of MTX in the urine of the patients of diffuse large B cell lymphoma was also demonstrated.

SIMULTANEOUS DETERMINATION OF 14 QUINOLONES IN ROYAL JELLY BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY USING ANION-EXCHANGE SOLID-PHASE EXTRACTION

A new method based on liquid chromatography-tandem mass spectrometry has been developed for simultaneous determination of 14 quinolones (QNs) residues, including ciprofloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, flumequine, lomefloxacin, marbofloxacin, norfloxacin, orbifloxacin, ofloxacin, pipemidic acid, pefloxacin, and sarafloxacin in royal jelly. The proposed analytical procedure involves extraction of the QNs from samples by 0.1 M sodium hydroxide aqueous solution, a step for clean-up and preconcentration of the analytes by anion-exchange solid-phase extraction (SPE) and liquid chromatographic separation with mass spectrometric detection. Internal standard calibration was applied with norfloxacin-D5 as internal standard (I.S.). Satisfactory recovery (from 64% to 113%), excellent repeatability precision (relative standard deviations, RSDs below 6%), reproducibility precision (RSDs below 10%), and low limits of quantifications (LOQs from 0.3 to 2.5 µg kg−1) were evaluated from spiked royal jelly samples at 2.5, 5.0, and 10.0 µg kg−1 three concentration levels. This protocol has been validated by five authoritative laboratories and successfully used for analysis of a large number of import–export samples (n > 2000). Norfloxacin and ciprofloxacin were found to be the most common contamination in royal jelly in the ranges of 3.5–30 µg kg−1 and 4.5–20 µg kg−1, respectively.

Molecular mass spectrometric identification of superoxide dismutase in the liver of mice Mus musculus and Mus spretus using a metallomics analytical approach

This paper reports the identification and quantification of superoxide dismutase in the liver of Mus musculus and Mus spretus mice using a metallomics analytical approach. The approach consisted of using orthogonal chromatographic systems coupled to ICP-MS and UV detectors. Size-exclusion fractionation of the cytosolic extracts was followed by anion-exchange chromatographic separation of Cu- and Zn-containing species. After purification then tryptic digestion, Cu- and Zn-containing superoxide dismutase was identified by nESI-QqTOF. The MS-MS spectra of doubly charged peptides, with the Mascot searching engine, were used to obtain the sequence of the protein.

Anion-Exchange Chromatographic Separation of Hg for Isotope Ratio Measurements by Multicollector ICPMS

A procedure is described for precise Hg isotope ratio measurements by solution nebulization multicollector inductively coupled plasma mass spectrometry (MC-ICPMS). Hg was released from geological samples using aqua regia extraction and then separated from other matrix elements with the aid of anion-exchange chromatography using strongly basic Dowex 1-X8 anion-exchange resin. Performance of the chromatographic procedure was evaluated using various types of replacement anions for elution of mercury, including l-cysteine, thiourea, NO3-, and SO42-. A solution of 0.15% l-cysteine in 0.06 M HCl was found to be the most convenient eluent for subsequent MC-ICPMS measurements. The optimized procedure provides separation of Hg from virtually all concomitant matrix elements while maintaining quantitative (>95%) recovery. In addition, band displacement chromatographic experiments were conducted to assess whether the anion-exchange purification can produce Hg isotope fractionation artifacts. No isotope fractionation between the Hg(II)-l-cysteine complex in aqueous solution and Hg ions in the anion-exchange resin was observed. Hg isotope ratio measurements were performed using the bracketing standards approach and on-line correction for instrumental mass discrimination using Tl spiking and normalization to the 205Tl/203Tl ratio. The absence of spectral interference during Hg isotope ratio measurements was verified using a three-isotope plot. Uncertainties of Hg isotope ratio measurements for replication of the entire procedure, expressed as two standard deviations, are better than +/-0.08 per thousand/amu. The described procedure facilitates study of variations in the isotopic composition of Hg in nature.

Stimulation of Mg 2+-independent form of Ca 2+-ATPase by a low molecular mass protein purified from goat testes cytosol

A low molecular mass protein purified from goat ( Capra hircus) testes cytosol following gel filtration and anion exchange chromatographic separation stimulates Mg 2+-independent Ca 2+-ATPase activity without any significant effect on Mg 2+-dependent Ca 2+-ATPase. Stimulation of the ATPase is due to an increase in the rate of dephosphorylation of the overall reaction step of the enzyme. Binding of the stimulator increases the affinity of Ca 2+-ATPase for Ca 2+. An analysis of enzyme kinetics reveals a reversible type of binding of the stimulator to the ATPase and non-competitive type of stimulation with respect to the substrate. Stimulation seems due to binding of the protein at a single site following Michaelis–Menten model. The protein can also counter the effect of calcium antagonists exerted on the ATPase. The pI of the protein is 6.2 and its molecular mass has been determined to be 13, 961 by Q-TOF-MS.

Comment on “On the use of anion exchange chromatography for the characterization of water soluble organic carbon” by H. Chang et al.

Comment on “On the use of anion exchange chromatography for the characterization of water soluble organic carbon” by H. Chang et al.

Determination of hexavalent chromium by using speciated isotope-dilution mass spectrometry after microwave speciated extraction of environmental and other solid materials

Precise and accurate determination of hexavalent chromium in different types of solid environmental sample is regarded as a technical challenge with significant potential error if historically accepted methods are used. Microwave-assisted alkaline extraction (0.5 mol L(-1) NaOH + 0.28 mol L(-1) Na2CO3) followed by anion-exchange chromatographic separation and inductively coupled plasma mass spectrophotometric detection has been shown to provide accurate and precise results. To obtain a better understanding of potential species conversion during and/or after extraction steps, speciated isotope-dilution mass spectrometry (SIDMS) (EPA Method 6800) metrology has been successfully applied as a diagnostic tool with the modified accompanying extraction version of EPA Method 3060A. In our study, aggregate materials distributed over a large area of a major western US state were found to contain a high concentration of total chromium (195 +/- 13 to 709 +/- 19 microg g(-1)) and significant amounts of Cr6+ (141 +/- 6 to 341 +/- 29 microg g(-1)) which are at least three orders of magnitude higher than the US EPA threshold limit (0.5 microg g(-1)). Sediment samples from a major western US state, studied independently, were found to contain less (1.77 +/- 0.34 microg g(-1)) or no Cr6+ in the presence of significant total chromium.

Sensitive quantification of iodide by ion-exchange chromatography with electrochemical detection at a modified platinum electrode

A rapid and very sensitive method for the accurate determination of free iodide in real samples is described. The method is based on anion-exchange chromatographic separation coupled with amperometric detection at a modified platinum electrode under constant applied potential (+0.85 V vs. Ag AgCl). An experimental setup with an in-line and very effective method of electrode modification is proposed using an amperometric thin-layer cross-flow detector and a flowing solution 300 mg/L of iodide; the working electrode is polarised to the limiting current for oxidation of iodide to iodine in acidic solutions with the consequent formation of an iodine-based film. The results indicated that the modified electrode exhibits high analytical response for iodide electrooxidation with good stability and long-life. The signal intensity of daily experimental sessions (8 h), during which standards and real samples were repeatedly injected, exhibits a moderate lowering (i.e. < 6%). Using a mixture of 25 mM HNO3 and 50 mM NaNO3 as an eluent phase in ion-exchange chromatography, the detection limit of iodide was estimated to be 0.5 microg/L (S/N = 3) with an injection volume of 50 microL. This method was applied successfully to quantify the iodide content of milk samples and in wastewaters as well as trace amounts in common vegetables and solutions containing high chloride levels.

Study of the feasibility of using a pellicular anion-exchange column for separation of transferrin isoforms in human serum by HPLC with UV detection.

A method has been optimised for the separation of glycoforms of human serum transferrin, using a high-performance pellicular anion-exchange chromatographic column. The effect of the eluent pH and of the column temperature on the separation of transferrin glycoforms was studied using a standard solution of commercially available human serum transferrin. An HPLC system equipped with an ultraviolet detector was used for the analysis. No immunoassay was used after the anion-exchange chromatographic separation of the glycoforms, in contrast with most currently used methods. The method was applied to the separation and quantification of transferrin glycoforms in serum from a healthy, non-pregnant woman, after saturation of transferrin with iron and further precipitation of lipoproteins. The whole chromatographic run, including re-equilibration of the column, took 35 min.