EDTA In Aqueous Solution Research Articles

Dystrophic calcinosis: structural and morphological composition, and evaluation of ethylenediaminetetraacetic acid (‘EDTA’) for potential local treatment

BackgroundTo perform a detailed morphological analysis of the inorganic portion of two different clinical presentations of calcium-based deposits retrieved from subjects with SSc and identify a chemical dissolution of these deposits suitable for clinical use.MethodsChemical analysis using Fourier Transform IR spectroscopy (‘FTIR’), Raman microscopy, Powder X-Ray Diffraction (‘PXRD’), and Transmission Electron Microscopy (‘TEM’) was undertaken of two distinct types of calcinosis deposits: paste and stone. Calcinosis sample titration with ethylenediaminetetraacetic acid (‘EDTA’) assessed the concentration at which the EDTA dissolved the calcinosis deposits in vitro.ResultsFTIR spectra of the samples displayed peaks characteristic of hydroxyapatite, where signals attributable to the phosphate and carbonate ions were all identified. Polymorph characterization using Raman spectra were identical to a hydroxyapatite reference while the PXRD and electron diffraction patterns conclusively identified the mineral present as hydroxyapatite. TEM analysis showed differences of morphology between the samples. Rounded particles from stone samples were up to a few micron in size, while needle-like crystals from paste samples reached up to 0.5 µm in length.Calcium phosphate deposits were effectively dissolved with 3% aqueous solutions of EDTA, in vitro. Complete dissolution of both types of deposit was achieved in approximately 30 min using a molar ratio of EDTA/HAp of ≈ 300.ConclusionsStone and paste calcium-based deposits both comprise hydroxyapatite, but the constituent crystals vary in size and morphology. Hydroxyapatite is the only crystalline polymorph present in the SSc-related calcinosis deposits. Hydroxyapatite can be dissolved in vitro using a dosage of EDTA considered safe for clinical application. Further research is required to establish the optimal medium to develop the medical product, determine the protocol for clinical application, and to assess the effectiveness of EDTA for local treatment of dystrophic calcinosis.

Volumetric and acoustic characteristics of a temperature-dependent aqueous solution of trisodium EDTA with EG, DEG and TEG

The apparent molar volume(Vϕ) and apparent molar isentropic compressibility (Kϕ,s) of ethylene glycol (EG), diethylene glycol (DEG) and triethylene glycol (TEG) in trisodium ethylenediaminetetraacetic acid solution (aqueous) have been estimated based on measured experiments values for both the density (ρ) and sound speed (s) at (288.15, 298.15, 308.15 and 318.15) K and 0.01 MPa as well. From the foundation in infinite dilution limiting apparent molar volumes (Vϕ∘) and isentropic compressibility(Kϕ,s∘) its transfer properties (ΔVϕ∘, ΔKϕ,s∘), expansibility coefficients(∂Eϕ∘/∂T)P, expansion coefficient (α) and interactions coefficients (VAB,KAB,VABB,KABB) the various interactions among glycols and EDTA in aqueous solution were investigated and addressed. Fourier transform infrared spectroscopy study had also been carried in binary solution.

Volumetric-acoustic study of Disodium Ethylenediaminetetraacetic acid with Propylene and Hexylene glycol at different temperatures

The present study shows the molecular interactions at constant pressure of 0.1 megapascals (MPa) and a range of temperatures from (288.15, 298.15, 308.15, 318.15 K) in aqueous solution of Disodium EDTA in (0.01, 0.02, 0.03) mol.kg-1 along with Propylene glycol and Hexylene glycol. Speed of sound and density data is determined using Anton Paar DSA 5000 M. Various thermo-acoustic parameters are calculated using the obtained data. Experimental density values determined apparent Molar Volume (Vϕ), Partial Molar Volume (Vϕ°) as well as Partial Molar Volume of Transfer (ΔVϕ°) along with Partial Molar Expansibility (Eϕ°) and thermal expansion coefficient (α). While speed of sound determined the Apparent Molar Isentropic Compression (Kϕ,S), Partial Molar Isentropic Compression (Kϕ,S°) as well as Partial Molar Isentropic Compression of Transfer (ΔKϕ.S°). Pair and Triplet Coefficients along with Empirical constants have also been estimated. FTIR spectroscopic research had been carried out in the binary mixture. From propylene through hexylene glycols, these factors provide data related to significant interactions between solute and solvent.

Mercury removal from aqueous media by aminated polyacrylonitrile beads impregnated with nano-urchin bismuth sulfide particles: Experimental and theoretical analysis under batch and continuous mode

The present study encompasses a thorough analysis of aqueous Hg (II) adsorption using aminated polyacrylonitrile beads incorporated with high-capacity surfactant assisted bismuth sulfide nanoparticles. The porous beads provided effective contact between aqueous Hg and the inorganic nano-adsorbent (PBS2) that was encapsulated in the matrix. The macro-sized beads served as suitable packing material for designing a fixed bed adsorption column for continuous mode analysis without significant pressure build up and offered an easy solid–liquid separation after treatment. A marginal negative zeta potential of the adsorbent beads improved the interaction with Hg2+ facilitating the sorption. The incorporation of amine groups into the polymeric bead surface via amination increased the adsorption capacity further from 83.30 mg/g to 130.00 mg/g (∼56.00%) at 303 K. The associated parameters of batch kinetics (effective pore diffusivity and mass transfer coefficient) were determined using a first principle-based model available in literature. Mercury was chemisorbed via synergistic lone pair interactions of S and N atoms with Hg, leading to formation of coordination bonds (Hg-S and Hg-N). An established mass transfer model was utilized to generate simulated breakthrough curves for the continuous analysis using synthetic feed. Additionally, the adsorption column showed comparable performance using both synthetic and real water samples (synthetically spiked) maintaining high selectivity. The optimized values of axial dispersion coefficient (Ez) and fixed bed mass transfer coefficient (kfb) were estimated and subsequently used for the scale up studies to predict column life. The adsorbent beads were regenerated using 0.05 M EDTA aqueous solution and showed good reusability (>95.00%) after 5 cycles of operation. Overall, this work highlights a reusable Hg adsorbent possessing good selectivity and capacity and simultaneously demonstrating real-field applicability.

Critical Evaluation of Two Qualitative Analytical Approaches for Multiclass Determination of Veterinary Drugs in Bovine Muscle Using UHPLC-Q-Orbitrap: The Wind of Change in Brazilian Monitoring.

Food safety is recognized as a main requirement for consumers, food industries, and official laboratories. Here, we present the optimization and screening qualitative validation of two multianalyte methods in bovine muscle tissues by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry with an Orbitrap-type analyzer, operated with a heated ionization source in positive and negative mode. This aims for not only the simultaneous detection of veterinary drugs regulated in Brazil but also the prospection of antimicrobials not yet monitored. Two different sample preparation procedures were applied: method A-generic solid-liquid extraction with 0.1% formic acid (v/v) in an aqueous solution of EDTA 0.1% (w/v)-acetonitrile-methanol (1:1:1, v/v/v), followed by an additional ultrasound-assisted extraction and method B-QuEChERS. In both procedures, selectivity showed satisfactory conformity. From a detection capability (CCβ) equivalent to ½ the maximum residue limit, >34% of the analyte resulted in a false positive rate of <5%, preponderant by the QuEChERS method, which exhibited a higher yield of the sample. The results showed the potential application of both procedures in the routine analysis of foods by official laboratories, enabling the expansion of this methodological portfolio as well as its analytical scopes, thus optimizing the control of residues of veterinary drugs in the country.

Development of an Automatable Affinity Purification Process for DNA-Encoded Chemistry.

DNA-encoded library technologies require high-throughput, compatible, and well automatable platforms for chemistry development, building block rehearsal, and library synthesis. An affinity-based process using Watson–Crick interactions was developed that enables purification of DNA-tagged compounds from complex reaction mixtures. The purification relies on a single-stranded DNA-oligonucleotide, called capture strand, which was covalently coupled to an agarose matrix and to which a DNA-compound conjugate from a DNA-encoded library (DEL) reaction can be reversibly annealed to. The thus-formed DNA duplex tolerated surprisingly stringent washing conditions with multiple solvents to remove excess reactants and reagents. The tolerated solvents included aqueous buffers, aqueous EDTA solutions to remove metal ions, aqueous mixtures of organic solvents, and even pure organic solvents. The purified DNA-conjugate was eluted with aqueous ammonia and could be used for reaction analysis or for instance in DNA-encoded library synthesis. The lab equipment for purification was tailored for automation with open-source hardware and constructed by 3D printing.

Chelating Silicone Dendrons: Trying to Impact Organisms by Disrupting Ions at Interfaces.

The viability of pathogens at interfaces can be disrupted by the presence of (cationic) charge and chelating groups. We report on the synthesis of silicone dendrimers and linear polymers based on a motif of hexadentate ligands with the ability to capture and deliver metal ions. Mono-, di- or trialkoxysilanes are converted in G1 to analogous vinylsilicones and then, iteratively using the Piers-Rubinsztajn reaction and hydrosilylation, each vinyl group is transformed into a trivinyl cluster at G2. The thiol-ene reaction with cysteamine or 3-mercaptopropionic acid and the trivinyl cluster leads to hexadentate ligands 3 × N–S or 3 × HOOC–S. The compounds were shown to effectively capture a variety of metals ions. Copper ion chelation was pursued in more detail, because of its toxicity. On average, metal ions form chelates with 2.4 of the three ligands in a cluster. Upon chelation, viscous oils are converted to (very) soft elastomers. Most of the ions could be stripped from the elastomers using aqueous EDTA solutions, demonstrating the ability of the silicones to both sequester and deliver ions. However, complete ion removal is not observed; at equilibrium, the silicones remain ionically crosslinked.

A novel polymeric hybrid sensory smart material for the prompt recognition of mercury ions in water

An acrylamide-xanthene hybrid polymeric probe was synthesized for the selective and sensitive detection of mercury ions (Hg2+) in 100% water. The non-polymeric form of probe displayed extraordinary selectivity above other additional metal cations via both a visual color change and fluorescence change with exceptional reversibility and responds rapidly to changes in the environment. The 1H NMR titration study revealed the binding of an Hg2+ ion on the imine hydrogen of the probe by the opening of the spiro-lactam ring, which instigates an instant color change. Reversible behavior of the polymeric probe using an aqueous solution of EDTA was confirmed by both absorption and fluorescence studies. The prepared polymeric probe detects Hg2+ in 100% water and is a promising material for the mercury recognition in real-time.

An ion chromatography tandem mass spectrometry (IC-MS/MS) method for glyphosate and amino methyl phosphoric acid in serum of occupational workers

A non-suppressed ion chromatography tandem mass spectrometry (IC-MS/MS) has been established to measure glyphosate (GLY) and amino methyl phosphoric acid (AMPA) in serum of the occupational workers. After the inhibition of metal-chelating with EDTA aqueous solution and protein precipitation with acetonitrile, the samples were purified with a modified liquid–liquid extraction method and separated on an ion chromatographic column (250 mm × 2.1 mm) by using a gradient program of 0.4 mol/L methylamine aqueous solution and acetonitrile as mobile phase. Multiple reactions monitoring (MRM) model was used to obtain the sensitivity of the method, and the isotopically labelled internal standards were applied in the calibration process. Good linearities and recoveries were obtained in the validation process. The recovery experiment provided satisfied results with a range of 91%~101% for GLY and 92%~99% for AMPA in human serum and RSD values for both two analytes were <4%. The limits of detection for GLY and AMPA were 0.6 ng/mL and 1.2 ng/mL, respectively. The association between the posts in workplace and the concentration of two contaminants in serum samples was investigated throughout the analysis of the 307 worker serum samples. The data analysis results indicated the distribution of compounds in human serum was significantly related to the exposure probability and duration during the manufacture process of GLY.

Study of Ho doped Ag2S thin films prepared by CBD method

Abstract Ho doped Ag2S thin films were grown on the glass substrate by chemical bath deposition (CBD) method at room temperature. The bath contained aqueous solution of silver nitrate, thiourea, EDTA, ammonia and holmium nitrate. Silver nitrate was used as a silver (Ag+) ion source; thiourea as sulfur (S−) ion source, EDTA was a complexing agent while ammonia was used to maintain pH, Ho(NO3)3 was taken as a source of Ho ions. The optical absorption edge of undoped (pure) and Ho doped silver sulfide films was determined between 324 nm and 298 nm showing blue shift as compared to bulk Ag2S. Band gaps calculated from Tauc plot also showed an increase in values for doped samples. The increase in band gap indicates reduced particle size in the prepared Ho doped films. The photoluminescence emission peaks were observed at around 578 nm to 601 nm wavelength and excitation peaks were found at 351 nm to 294 nm for undoped and doped films. The SEM micrograph consists of globular ball type and flower type structures observed in the prepared films of Ho doped Ag2S.

Isotopic fingerprints indicate distinct strategies of Fe uptake in rice

Plants typically take up Fe through either strategy I or strategy II, whereas rice uses both. Stable Fe isotopes potentially reveal pathways of Fe uptake in rice plants. In this study we measured Fe isotopic compositions of rice grown under different conditions, i.e. paddy soil with deficient Fe supply and Fe3+-EDTA aqueous solution with sufficient Fe supply, to investigate whether Fe isotope fractionation is distinct under Fe-deficient and Fe-sufficient conditions as well as their possible controlling mechanisms. Our results show that rice grown in the Fe3+-EDTA aqueous solutions with sufficient Fe supply preferentially takes up light Fe isotopes (Δ56Febulk plant-nutrient = −1.36‰) and accumulates Fe in different parts of plant (roots, stems, leaves, husks, and grains) with large Fe isotope fractionation. Under such a growing condition, rice takes up Fe through strategy I and the reduction of Fe3+ to Fe2+ results in plants enriching isotopically light Fe. Within the plant, the transportation of Fe is accompanied with changes in redox state, which thus causes significant Fe isotope fractionation among different plant tissues. In contrast, rice plants grown in soils with deficient Fe supply are slightly enriched in heavy Fe isotopes (Δ56Febulk plant-soil solution = 0.27‰) and accumulate Fe in different plant tissues with little isotope fractionation. Under this growing condition, the rice plant takes up Fe through strategy II. Plants take up Fe from soils as Fe3+-phytosiderophores (Fe3+-PS) complex and transport Fe as Fe3+-nicotianamine (Fe3+-NA) complex throughout the plant, which does not involve changes in redox state and thus results in very limited isotope fractionation. The observed difference in Fe isotope fractionation indicates two distinct Fe uptake and translocation strategies for rice grown under the two different conditions: strategy I under Fe-sufficient conditions, and strategy II under Fe-deficient conditions. Our results demonstrate that Fe isotope ratios may be used to distinguish Fe-sufficient versus Fe-deficient conditions and to elucidate Fe biogeochemical processes during rice growth.

Determination of p-Phenylene diamine in workplace air by elution solution-liquid chromatography

Objective: To establish a elution solution-liquid chromatography method for determination of p-Phenylene diamine (PPD) in workplace air. Methods: p-Phenylene diamine (PPD) in the air of workplace was collected with glass fiber filters coated with dilute sulfuric acid and extracted with an aqueous EDTA solution. The target toxicant was separated with the C(18) column and analyzed with UV detector, identified by retention time, and quantified by peak area. Results: The linear range of PPD in the air of workplace was 2.00~10.00 μg/ml, with a correlation coefficient of 0.999 96. The limit of detection was 0.07 μg/ml. The lower limit of quantification was 0.23 μg/ml. The minimum detectable concentration was 0.003 1 mg/m(3) under 45.0 L sampling volume and 2.0 ml extraction solution volume. The within-run precision of different PPD concentrations was 0.15%~2.3% and the between-run precisions was 1.4%~2.6%; The extraction efficiencies was 91.4%~95.4%; The average collection efficiencies was 96.6%; The samples could be stored for 7 days isolation of air. The potential coexistence of m-Phenylene diamine and o-Phenylene diamine with p-Phenylene diamine (PPD) in the air did not interfere with the results of determination. Conclusion: This method has high sensitivity, precision, accuracy and lower limit of detection and it is applicable for determination of p-Phenylene diamine (PPD) in workplace air.

Sonocatalytic oxidation of EDTA in aqueous solutions over noble metal-free Co3O4/TiO2 catalyst

The sonocatalytic degradation of EDTA in aqueous solution was studied under ultrasound irradiation (345 kHz, 73 W, acoustic power 0.20 W·mL−1, Ar and Ar/O2 saturating gases, T = 20–50 °C) in the presence of Co3O4/TiO2 and Pt/TiO2 nanocatalysts. About 90% of EDTA (C0 = 5 10-3 M) was oxidized during ultrasonic treatment at 40 °C in the presence of the Co3O4/TiO2 catalyst and Ar/O2 gas mixture. By contrast, Pt/TiO2 catalyst exhibited much lower sonocatalytic activity in this system. Suggested mechanism of EDTA oxidation in the presence of Co3O4/TiO2 catalyst involved the generation of oxidizing radicals by acoustic cavitation and Co(II)-Co(III) redox process. Quite low apparent activation energy of the sonocatalytic process (Ea = 19 kJ mol−1) was attributed to diffusion of reagents in the vicinity of the active sites of catalyst. Sonocatalytic degradation of EDTA is accompanied by formation of iminodiacetic acid, formic acid, oxalic acid, glycolic acid and acetic acid as intermediate products in an agreement with radical-driven mechanism.

Novel Task Specific Ionic Liquids to Remove Heavy Metals from Aqueous Effluents

Task Specific Ionic Liquids (ILs) were generated by association between a cationic ester derivative of betaine and coordinating inorganic anions such as dicyanamide (Dca−), chlorosalycilate (ClSal) and saccharinate (sac). Extraction of Cu(II), Ni(II), Co.(II), Pb(II) and Cd(II) from water was performed with these ILs at room temperature. Our results show that ionic liquid with Clsal anions have a high extraction efficiency towards Cu(II), Ni(II), Cd(II), and Pb(II), whereas dicyanamide ionic liquid may extract efficiently Cu(II), Ni(II) Co.(II) and Cd(II). Ionic liquids with saccharinate anions are selective of Cd(II) ions. The extraction mechanism has been studied by the determination of the coextraction of the counter ion of the metal salt. Our results show that the extraction mechanism proceeds via a mixed process involving both cation exchange and ion-pairing. The proportion of which depends on the nature of the cation. The coordination of Cu(II), Ni(II) and Co.(II) in ionic liquid phase was followed by UV-vis spectroscopies. The metal could be back-extracted from the ionic liquid phase with aqueous EDTA solutions. The metal extractability of the ionic liquid after the back-extraction is equivalent to that of the fresh mixture showing that ionic liquid can be reused for several extraction and back-extraction cycles.

Amphiphilic mesoporous graphene mediated efficient photoionic cell

Photoionic cell is a facile concept for direct solar energy conversion and storage based on dye-quencher couples. We propose a system based on dye modified amphiphilic mesoporous graphene (APG) and cobalt EDTA as a photoionic cell. Upon light irradiation, the dyes are excited and quenched by cobalt EDTA in aqueous solution to form oxidized cobalt EDTA and reduced dyes that are hydrophobic. Solar energy is then converted into electrochemical energy in the form of the oxidized cobalt EDTA in aqueous solutions and reduced dyes in APG. Compared to organic phase usually used for photoionic cells, the usage of APG can lead to increased amount of dye molecules in the photoionic cells, and can achieve approximately 100% separation of the reduced dyes from the oxidized cobalt EDTA for long term energy storage.

Multiclass method for the determination of 62 antibiotics in milk.

A multiclass method for screening and confirmatory analysis of antimicrobial residues in milk has been developed and validated. Sixty-two antibiotics belonging to ten different drug families (amphenicols, cephalosporins, lincosamides, macrolides, penicillin, pleuromutilins, quinolones, rifamycins, sulfonamides and tetracyclines) have been included. After the addition of an aqueous solution of EDTA, the milk samples were extracted twice with acetonitrile, evaporated and dissolved in ammonium acetate. After centrifugation, 10 µl were analysed using LC-Q-Orbitrap operating in positive electrospray ionization mode. The method was validated in bovine milk in the range 2-150 µg kg(-1) for all antibiotics; for four compounds with maximum residue limits higher than 100 µg kg(-1) , the validation interval has been extended until 333 µg kg(-1) . The estimated performance characteristics were satisfactory complying with the requirements of Commission Decision 2002/657/EC. Good accuracies were obtained also taking advantage from the versatility of the hybrid mass analyser. Identification criteria were achieved verifying the mass accuracy and ion ratio of two ions, including the pseudomolecular one, where possible. Finally, the developed procedure was applied to 13 real cases of suspect milk samples (microbiological assay) confirming the presence of one or more antibiotics, although frequently, the maximum residue limits were not exceeded. The availability of rapid multiclass confirmatory methods can avoid wastes of suspect, but compliant, raw milk samples. Copyright © 2016 John Wiley & Sons, Ltd.

고온화학세정환경에서 20 % EDTA 용액이 결함 전열관 (Alloy600)에 미치는 영향

The transport and deposition of corrosion products in pressurized water nuclear reactor (PWR) steam generators have led to corrosion (SCC, denting etc.) problems. Lancing, mechanical cleaning and chemical cleaning have been used to reduce these problems. The methods of lancing and mechanical cleaning have limitations in removing corrosion products due to the structure of steam generator tubes. But high temperature chemical cleaning (HTCC) with EDTA is the most effective method to remove corrosion products regardless of the structure. However, EDTA in chemical cleaning aqueous solution and chemical cleaning environments affects the integrity of materials used in steam generators. The nuclear power plants have to perform the pre-test (also called as qualification test (QT)) that confirms the effect on the integrity of materials after HTCC. This is one of the series studies that assess the effect, and this study determines the effects of 20 % EDTA aqueous solution on defective tubes in high temperature chemical cleaning environments. The depth and magnitude of defects in steam generator (SG) tubes were measured by eddy current test (ECT) signals. Surface analysis and magnitude of defects were performed by using SEM/EDS. Corrosion rate was assessed by weight loss of specimens. The ECT signals (potential and depth %) of defective tubes increased marginally. But the lengths of defects, oxides on the surface and weights of specimens did not change. The average corrosion rate of standard corrosion specimens was negligible. But the surfaces on specimens showed traces of etching. The depth of etching showed a range on the nanometer. After comprehensive evaluation of all the results, it is concluded that 20 % EDTA aqueous solution in high temperature chemical cleaning environments does not have a negative effect on defective tubes.

Screening and confirmatory method for multiclass determination of 62 antibiotics in meat

A multiclass method for screening and confirmatory analysis of antimicrobial residues in muscle has been developed and validated, according to Commission Decision 2002/657/EC. Sixty-two antibiotics belonging to ten different drug families (amphenicols, beta-lactams, diamino-pyrimidine, lincosamides, macrolides, pleuromutilins, quinolones, rifamycins, sulfonamides and tetracyclines) have been included in the method. After the addition of an aqueous solution of EDTA, the minced muscle was extracted with acetonitrile/water mixture and, later, with pure acetonitrile. The extract was evaporated and redissolved in ammonium acetate buffer prior to LC injection. Instrumental determination was performed by liquid chromatography coupled to hybrid high resolution mass analyser (LC-HRMS/MS) operating in positive electrospray ionization mode. Chromatographic separation was optimized on a Poroshell 120 EC-C18 column (100×3.0mm, 2.7μm) with gradient using methanol and water containing 0.1% of formic acid as mobile phases. The method was validated in bovine muscle in the range 3.3–150μgkg−1 for all antibiotics; for some compounds with MRL higher than 100μgkg−1, the validation interval has been extended until 1500μgkg−1. The studied performance characteristics were selectivity, linearity, precision, trueness (recovery), decision limits, detection capabilities, detection and quantification limits. Satisfactory quantitative performances were obtained for all the analytes. Ruggedness tests demonstrated the applicability to swine and poultry muscle, too. Finally the wide participation in proficiency tests allowed to investigate in deep the method performances.

Utilization of milk protein as an environmental material: accumulation of metal ions using a protein–inorganic hybrid material

A casein–inorganic hybrid material was prepared by mixing casein, one of phosphoproteins, and a silane coupling reagent. This casein–inorganic hybrid material was stable in an aqueous solution owing to the electrostatic interactions between the phosphate group in casein and the amino group in the silane coupling reagent. The hybrid materials could selectively accumulate heavy and light metal ions and did not interact with the rare-earth metal ions. In addition, the metal ion-accumulated hybrid materials could be recycled by washing with an aqueous EDTA solution or hydrochloric acid.

Short-column anion-exchange chromatography for soil and peat humic substances profiling by step-wise gradient of high pH aqueous sodium ethylenediaminetetraacetate

Novel anion-exchange liquid chromatographic method with step gradient of aqueous EDTA4− based mobile phase elution has been developed to profile available Slovak soil humic substances and alkaline extracts of various soils. The method utilize short glass column (30mm×3mm) filled in with hydrolytically stable particles (60μm diameter) Separon HEMA-BIO 1000 having (diethylamino)ethyl functional groups. Step gradient was programmed by mixing mobile phase composed of aqueous solution of sodium EDTA (pH 12.0; 5mmolL−1) and mobile phase constituted of aqueous solution of sodium EDTA (pH 12.0, 500mmolL−1). The FLD of HSs was set to excitation wavelength 480nm and emission wavelength 530nm (λem). Separation mechanism was studied by use of selected aromatic acids related to humic acids with the aid of UV spectrophotometric detection at 280nm. The proposed method benefits from high ionic strength (I=5molL−1) of the end mobile phase buffer and provides high recovery of humic acids (98%). Accurate and reproducible profiling of studied humic substances, alkaline extracts of various types of soils enables straightforward characterization and differentiation of HSs in arable and forest soils. Selected model aromatic acids were used for separation mechanism elucidation.