强碱阴离子交换树脂及碱性洗脱剂的离子排阻/阴离子交换色谱法同时测定NH4+ ，NO2-和NO3-（英文）

Ion-exclusion/anion-exchange chromatography(IEC/AEC) on a combination of a strongly basic anion-exchange resin in the OH——form with basic eluent has been developed.The separation mechanism is based on the ion-exclusion/penetration effect for cations and the anion-exchange effect for anions to anion-exchange resin phase.This system is useful for simultaneous separation and determination of ammonium ion(NH+4),nitrite ion(NO-2),and nitrate ion(NO-3) in water samples.The resolution of analyte ions can be manipulated by changing the concentration of base in eluent on a polystyrene-divinylbenzene based strongly basic anion-exchange resin column.In this study,several separation columns,which consisted of different particle sizes,different functional groups and different anion-exchange capacities,were compared.As the results,the separation column with the smaller anion-exchange capacity(TSKgel Super IC-Anion) showed well-resolved separation of cations and anions.In the optimization of the basic eluent,lithium hydroxide(LiOH) was used as the eluent and the optimal concentration was concluded to be 2 mmol/L,considering the resolution of analyte ions and the whole retention times.In the optimal conditions,the relative standard deviations of the peak areas and the retention times of NH+4,NO-2,and NO-3 ranged 1.28%-3.57% and 0.54%-1.55%,respectively.The limits of detection at signal-to-noise of 3 were 4.10 μmol/L for NH+4,1.87 μmol/L for NO-2 and 2.83 μmol/L for NO-3.     

Vacancy ion-exclusion chromatography of carboxylic acids on a weakly acidic cation-exchange resin

In this preliminary study, a new approach to ion-exclusion chromatography is proposed to overcome the relatively poor conductivity detection response which occurs in ion-exclusion chromatography when acids are added to the eluent in order to improve peak shape. This approach, termed vacancy ion-exclusion chromatography, requires the sample to be used as eluent and a sample of water to be injected onto a weakly acidic cation-exchange column (TSKgel OApak-A). Vacancy peaks for each of the analytes appear at the retention times of these analytes. Highly sensitive conductivity detection is possible and sharp, well-shaped peaks are produced, leading to efficient separations. Retention times were found to be affected by the concentration of the analytes in the eluent, and also by the presence of an organic modifier such as methanol in the eluent. Detection limits for oxalic, formic, acetic, propionic, butyric and valeric acids were 0.1, 0.2, 0.3, 0.3, 0.4 and 0.5 microM, respectively, and linear ranges for some acids extended over two orders of magnitude. Precision values for retention times were 0.21% and for peak areas were <1.90%. The vacancy ion-exclusion chromatography method was found to give detection responses four to 10 times higher than conventional ion-exclusion chromatography using sulfuric acid eluent and two to five times higher than conventional ion-exclusion chromatography using benzoic acid eluent.     

Prediction of the effects of methanol and competing ion concentration on retention in the ion chromatographic separation of anionic and cationic pharmaceutically related compounds

The mixed-mode separation of a selection of anionic and cationic pharmaceutically related compounds is studied using ion-exchange columns and eluents consisting of ionic salts (potassium hydroxide or methanesulfonic acid) and an organic modifier (methanol). All separations were performed using commercially available ion-exchange columns and an ion chromatography instrument modified to allow introduction of methanol into the eluent without introducing compatibility problems with the eluent generation system. Isocratic retention prediction was undertaken over the two-dimensional space defined by the concentration of the competing ion and the percentage of organic modifier in the eluent. Various empirical models describing the observed relationships between analyte retention and both the competing ion concentration and the percentage of methanol were evaluated, with the resultant model being capable of describing the separation, including peak width, over the entire experimental space based on six initial experiments. Average errors in retention time and peak width were less than 6% and 27%, respectively, for runs taken from both inside and outside of the experimental space. Separations performed under methanol gradient conditions (while holding the competing ion concentration constant) were also modelled. The observed effect on retention of varying the methanol composition differed between analytes with several analytes exhibiting increased retention with increased percentage methanol in the eluent. An empirical model was derived based on integration of the observed t_R vs. %methanol plot for each analyte. A combination of the isocratic and gradient models allowed for the prediction of retention time using multi-step methanol gradient profiles with average errors in predicted retention times being less than 4% over 30 different 2- and 3-step gradient profiles for anions and less than 6% over 14 different 2- and 3-step gradient profiles for cations. A modified peak compression model was used to estimate peak widths under these conditions. This provided adequate width prediction with the average error between observed and predicted peak widths being less than 15% for 40 1-, 2- and 3-step gradients for anions and less than 13% over 14 1-, 2- and 3-step gradients for cations.     

Influence of acidic eluent for retention behaviors of common anions and cations by ion-exclusion/cation-exchange chromatography on a weakly acidic cation-exchange resin in the H+ -form

Influence of acidic eluent on retention behaviors of common anions and cations by ion-exclusion/cation-exchange chromatography (ion-exclusion/CEC) were investigated on a weakly acidic cation-exchange resin in the H(+)-form with conductivity. Sensitivities of analyte ions, especially weak acid anions (F(-) and HCOO(-)), were affected with degree of background conductivity level with pK(a1) (first dissociation constant) of acid in eluent. The retention behaviors of anions and cations were related to that of elution dip induced after eluting acid to separation column and injecting analyte sample. These results were largely dependent on the natures of acid as eluent. Through this study, succinic acid as the eluent was suitable for simultaneous separation of strong acid anions (SO(4)(2-), Cl(-), NO(3)(-) and I(-)), weak acid anions (F(-), HCOO(-) and CH(3)COO(-)), and cations (Na(+), K(+), NH(4)(+), Mg(2+) and Ca(2+)). The separation was achieved in 20 min under the optimum eluent condition, 20 mM succinic acid/2 mM 18-crown-6. Detection limits at S/N=3 ranged from 0.10 to 0.51 microM for strong acid anions, 0.20 to 5.04 microM for weak acid anions and 0.75 to 1.72 microM for cations. The relative standard deviations of peak areas in the repeated chromatographic runs (n=10) were in the range of 1.1-2.9% for anions and 1.8-4.5% for cations. This method was successfully applied to hot spring water containing strong acid anions, weak acid anions and cations, with satisfactory results.     

Anion-exchange chromatography on short reversed-phase columns modified with amphoteric (N-dodecyl-N,N-dimethylammonio)alcanoates

Short reversed-phase columns (50 mm x 4.6 mm Gemini C(18)) were dynamically coated with carboxybetaines of the general structure, C(12)H(25)N(+)(CH(3))(2)(CH(2))(n)COOH, namely (N-dodecyl-N,N-dimethylammonio)undecanoate, DDMAU (n=10) and (N-dodecyl-N,N-dimethylammonio)butyrate, DDMAB (n=3), and investigated for the separation of inorganic anions in ion chromatography. The role of the ionic strength of coating surfactant solutions on their adsorption and resultant column capacity was studied. The retention of inorganic anions was investigated with different eluents at various concentrations and pH. Interestingly, no retention for anions was found with pure water as the eluent, but the addition of small amounts of electrolytes, up to 0.1 mM, caused a sharp increase in the retention of analytes. The effect of increasing anion retention with an increase in eluent cation charge was also observed. Based on this effect a new cation charge gradient concept was proposed and applied to the separation of a standard mixture of anions.     

A study of retention and overloading of basic compounds with mixed-mode reversed-phase/cation-exchange columns in high performance liquid chromatography

The retention and overload of bases were studied on two new mixed-mode, silica based phases possessing ionic carboxylate functionalities of different acidity embedded within a hydrophobic ligand (SiELC Primesep). At low pH, good peak shapes were obtained for small solute mass, suggesting that the mere presence of a mixed-mode hydrophobic/ionic retention mechanism is not responsible for the poor peak shape that can occur on conventional reversed-phases with ionised silanols. Somewhat inferior, but still acceptable peak shape for bases was obtained on a column containing a mixture of discrete ion exchange and reversed-phase particles (Hypersil Duet). In both types of column, the ionic sites favourably increased the capacity for ionised bases, reducing considerably the deterioration of peak shape with load observed with conventional RP columns. The combined ionic and reversed-phase interaction can give strong retention of bases under certain conditions, necessitating careful choice of stationary and mobile phase.     

On-line concentration of s-triazine herbicides in micellar electrokinetic chromatography using a cationic surfactant

A new method for the simultaneous determination of anions (sulfate, nitrate, and chloride) and cations (sodium, ammonium, potassium, magnesium, and calcium) in acid rain waters was investigated using high-performance ion-exclusion/cation-exchange chromatography with conductimetric detection on a separation column packed with a polymethacrylate-based weakly acidic cation-exchange resin in the hydrogen-form and an eluent comprising 1.5 mM sulfosalicylic acid–6 mM 18-crown-6 at pH 2.6, operated at 1.5 ml/min. Effective separation and highly sensitive conductimetric detection for the anions and the cations was achieved in about 14 min. Since the ionic balance (equivalents of anions/equivalents of cations) of acid rain waters of different pH (4.40–4.67) ranged from 0.97 to 0.94, evaluation of the water quality of acid rain was possible. This method was successfully applied to the simultaneous determination of the anions and the cations in acid rain transported from mainland China and North Korea to central Japan monitored by a meteorological satellite data analyzer.     

High-performance liquid chromatographic analysis of basic drugs on silica columns using non-aqueous ionic eluents. I. Factors influencing retention, peak shape and detector response

The use of silica columns together with non-aqueous ionic eluents provides a stable yet flexible system for the high-performance liquid chromatographic analysis of basic drugs. At constant ionic strength, eluent pH influences retention via ionisation of surface silanols and protonation of basic analytes, pKa values indicating the pH of maximum retention. At constant pH, retention is proportional to the reciprocal of the eluent ionic strength for fully protonated analytes and quaternary ammonium compounds. The addition of water up to 10% (v/v) has little effect on retention if the protonation of the analytes is unaffected. Thus, it is likely that retention is mediated primarily via cation exchange with surface silanols. However, additional factors must play a part with compounds such as morphine which give tailing peaks at acidic or neutral eluent pHs.     

Separation of -amino acids using a series of zwitterionic sulfobetaine exchangers

A set of five new covalently bond sulfobetaine exchangers with inner quaternary amines and outer sulfonic acids have been prepared by attachment of a series of zwitterionic precursors to hyperporous divinylbenzene polymers using a grafting reaction. The series of zwitterionic exchangers have the same backbone and identical spacers to the polymeric backbone, as well as comparable capacities. The only difference is the chain length for one to five methylene groups between the charged functional groups. Chromatographic properties are examined by separation of α-amino acids using sodium acetate and nitric acid eluents. The separation mechanism is explored by varying eluent ionic strength and eluent pH, resulting in the conclusion that amino acids are separated due to cation exchange interactions. This is a behavior never before observed using zwitterionic exchangers. It contradicts the fact that sulfobetaine-type materials used in zwitterionic ion chromatography (ZIC) usually are well suited for anion separation and only poorly for cation separation. Contrary to anion separations using the identical set of exchangers, the materials with three and four methylene groups between the charges give the highest retention factors. Materials showing the high potential in ZIC separations of inorganic anions give low retention factors for amino acids and vice versa.     

Zwitterionic ion chromatography with carboxybetaine surfactant-coated particle packed and monolithic type columns

Both particle packed (25 cm x 0.46 cm I.D. SUPELCOSIL 5 microm C18) and monolithic type (10 cm x 0.46 cm I.D. Merck Chromolith Performance C18) reversed-phase substrates were dynamically coated with a carboxybetaine type zwitterionic surfactant ((dodecyldimethyl-amino) acetic acid) and investigated as stationary phases for use in zwitterionic ion chromatography (ZIC). Investigations into eluent concentration and pH were carried out using KCl eluents containing 0.2 mM of the carboxybetaine surfactant to stabilise the column coatings. It was found that eluent concentration decreased anion retention whilst simultaneously increasing peak efficiencies, which may be due to the dissociation of intra- and inter-molecular salts of the carboxybetaine surfactant under higher ionic strength conditions. The Effect of eluent pH was an increase in anion retention with decreased eluent pH due to the increased protonation of the weak acid terminal group of the carboxybetaine, causing both a relative increase in the positive charge of the stationary phase and less repulsion of the anions by the dissociated weak acid group. The carboxybetaine-coated monolithic phase was applied to rapid anion separations using elevated flow rates and flow rate gradients.     

Simultaneous ion-exclusion/cation-exchange chromatography of anions and cations in acid rain waters on a weakly acidic cation-exchange resin by elution with sulfosalicylic acid

A simple, selective, and sensitive method for the simultaneous determination of anions (sulfate, nitrate, and chloride) and cations (sodium, ammonium, potassium, magnesium, and calcium) in acid rain waters was developed using ion-exclusion/ cation-exchange chromatography with conductimetric detection. A weakly acidic cation-exchange resin column (Tosho TSKgel OA-PAK-A) and a sulfosalicylic acid-methanol-water eluent was used. With a mobile phase comprising 1.25 mM sulfosalicylic acid in methanol-water (7.5:92.5) at 1.2 ml/min, simultaneous separation and detection of the above anions and cations was achieved in about 30 min. Linear calibration plots of peak area versus concentration were obtained over the concentration ranges 0-1.0 mM for anions (R=0.9991) and 0-0.5 mM for cations (R=0.9994). Detection limits calculated at S/N=3 ranged from 4.2 to 14.8 ppb for the anions and from 2.4 to 12.1 ppb for the cations. The reproducibility of retention times was 0.14-0.15% relative standard deviation (RSD) for anions and 0.18-0.31% for cations, and reproducibility of chromatographic peak areas was 1.22-1.75% RSD for anions and 1.81-2.10% for cations. The method was applied successfully to the simultaneous determination of anions and cations in aerosols transported from mainland China to central Japan, as determined by a meteorological satellite data analyzer.     

Separation of inorganic anions using a series of sulfobetaine exchangers

A set of five new sulfobetaine exchangers with inner quaternary amines and outer sulfonic acids have been prepared. A series of zwitterionic precursors was attached to highly porous divinylbenzene polymer using a grafting reaction, which allows a flexible adjustment of the degree of functionalisation. The resulting materials have identical spacers to the polymeric backbone and differ only in chain length between the charged functional groups. Capacities of the stationary phases were analysed by two different methods based on elemental analysis and the results obtained were found to correlate. The application of combustion elemental analysis proved an identical molar content of sulfur and nitrogen. The distance between the charged functional groups is varied from one to five methylene groups for a better understanding of the retention behaviour of inorganic anions on zwitterionic stationary phases. Inorganic anions were separated using sodium acetate eluents with varying ionic strength and pH and the behaviour of all columns has been compared to each other and to ZIC-HILIC and ZIC-pHILIC columns from Merck SeQuant. The exchangers with two and five methylene groups between the charges showed the highest retention factors. Polarity and accessibility of the anion exchange sites are dependent on the distance between the charged groups and the flexibility of the chains. These properties have a strong influence on anion separations. The exchanger with two methylene groups between the charged functional groups showed the biggest difference as compared to the commercially available ZIC-HILIC and ZIC-pHILIC exchangers.     

Chromatography of inorganic anions on poly(vinyl alcohol) gel columns with acidic eluents

Summary Poly(vinyl alcohol) (PVA) gel shows ionic retention properties for common inorganic anions when an acidic eluent is used. The ionic property of the PVA gel is due to the proton-acceptable nitrogen atoms of the cross-linking agent and the carboxylic residues being comprised in the gel matrix. The extent of the net charge on the gel surface depends on the pH of the eluent. At a pH ranging from 2.3 to 5.3, the PVA gel behaves as a weak anion exchanger with very low ion-exchange capacity. At these conditions four UV-absorbing inorganic anions (bromate, bromide, nitrate, and nitrite) are separated by eluting with aqueous sulfuric acid. Alkyl groups introduced on the gel surface hinder the ionized solute molecules from accessing to the positively charged functional groups on the gel surface. A neutral solute (HNO₂) is retained with non-ionic interactions.     

Vitamin U-bonded stationary phase in capillary ion chromatography

A vitamin U-bonded stationary phase was prepared and the retention behavior of inorganic anions was examined using ion chromatography. Inorganic anions were retained on the vitamin U-bonded stationary phase under acidic as well as neutral eluent conditions in the ion-exchange mode. The elution order of the examined anions under neutral eluent conditions was nearly the same as that observed in common ion exchange mode, while the elution order observed under acidic eluent conditions was completely different from that observed in common ion exchange mode. The retention of the analyte anions under the neutral eluent conditions was due to the sulfonium groups of the vitamin U, while protonated primary amino groups caused retention of the analyte anions with different selectivity under acidic conditions. The retention factor of the analyte anions increased with decreasing eluent concentration under both eluent conditions. The present system was applied to the determination of bromide and nitrate contained in seawater.     

Retention model for the separation of anionic metal-EDTA complexes in ion chromatography

A retention model is derived for complex anions eluted from an anion-exchange column with multiple ionic eluents containing hydrogencarbonate, carbonate, and hydroxyl species and the sample solution, containing transition metals, anions and complexing ligand. The theory is based on the generalized ion-exchange equilibrium, protonation and complex-formation equilibria. The unknown parameters of chromatographic ion-exchange equilibrium constants for sample and eluent species are determined from the experimental retention data by iterative minimization, using a non-linear regression algorithm. The model was utilized to predict the retention behaviour of CdEDTA²⁻, CoEDTA²⁻, MnEDTA²⁻ and NiEDTA²⁻ ions. The capacity factors of complex ions were determined for wide ranges of pH values and eluent concentrations. Good agreement was obtained between the observed and predicted retentions.     

Ion-exclusion chromatography with conductimetric detection of aliphatic carboxylic acids on a weakly acidic cation-exchange resin by elution with benzoic acid-beta-cyclodextrin

In this study, an aqueous solution consisting of benzoic acid with low background conductivity and beta-cyclodextrin (beta-CD) of hydrophilic nature and the inclusion effect to benzoic acid were used as eluent for the ion-exclusion chromatographic separation of aliphatic carboxylic acids with different pKa values and hydrophobicity on a polymethacrylate-based weakly acidic cation-exchange resin in the H+ form. With increasing concentration of beta-cyclodextrin in the eluent, the retention times of the carboxylic acids decreased due to the increased hydrophilicity of the polymethacrylate-based cation-exchange resin surface from the adsorption of OH groups of beta-cyclodextrin. Moreover, the eluent background conductivity decreased with increasing concentration of beta-cyclodextrin in 1 mM benzoic acid, which could result in higher sensitivity for conductimetric detection. The ion-exclusion chromatographic separation of carboxylic acids with high resolution and sensitivity was accomplished successfully by elution with a 1 mM benzoic acid-10 mM cyclodextrin solution without chemical suppression.     

阴离子交换树脂固相萃取分离全血中血红蛋白的研究

以330阴离子交换树脂为吸附材料建立了固相萃取分离血红蛋白的方法。利用血红蛋白和树脂之间的疏水作用力将血红蛋白吸附到树脂上,以Tris—HCl缓冲液（PH-8．9）为洗脱剂回收血红蛋白。考察了溶液PH值、吸附时间、离子强度、洗脱剂的种类及其酸度等对分离纯化效率的影响。在最优实验条件下,树脂对血红蛋白的吸附率和洗脱率分别为87％和70％,吸附容量为42．9μg／mg。吸收光谱和SDS—PAGE凝胶电泳证明,该方法可有效地从人全血中分离出纯度较高的血红蛋白。     

Retention of alkali,alkaline earth and transition metals on an itaconic acid cation-exchange column. Eluent pH,ionic strength and temperature effects upon selectivity

The unusual selectivity of a methylene succinic (itaconic) acid modified polymeric column was investigated for the separation of alkali, alkaline earth, transition and heavy metals employing non-chelating inorganic eluents. The retention of selected metal ions on the column was investigated with simple HNO3 eluents and eluents prepared from KNO3 and KCl salts of varying pH (adjusted using HNO3). From these studies both the effect of eluent ionic strength and pH upon retention was evaluated for the itaconic acid stationary phase. The results obtained showed that despite slow exchange kinetics causing poor efficiencies, acceptable baseline separations of selected alkaline earth and transitions could be obtained under optimum conditions (the baseline separation of Mg(II), Ca(II), Mn(II), Cd(II), Zn(II) and Co(II) was possible using a 15 mM KNO3-5 mM KCl eluent at pH 3.50 in under 25 min). The use of an simple ionic strength step gradient was shown that facilitated the addition of Pb(II) to the above group of metal ions. An investigation into the effect of temperature upon peak efficiency and retention showed increased column temperature could be used to improve the resolution of closely eluting metal ions such as Ca(II) and Sr(II) and Ca(II) and Mn(II).     

Double gradient ion chromatography using short monolithic columns modified with a long chained zwitterionic carboxybetaine surfactant

The rapid separation of inorganic anions on short monolithic columns permanently coated with a long chained zwitterionic carboxybetaine-type surfactant is shown. The surfactant, N-dodecyl-N,N-(dimethylammonio)undecanoate (DDMAU), was used to coat 2.5, 5.0 and 10 cm long reversed-phase silica monoliths, resulting in a permanent zwitterionic exchange surface when used with aqueous based eluents. The unique structure of the surfactant results in a charge double layer structure on the surface of the stationary phase, with strong internal anionic and weak external cationic exchange groups. The dissociation of the weak external carboxylic acid group acts to shield the inner anionic exchange site, resulting in substantial effective capacity changes with eluent pH. Utilising this effect with the application of an eluent pH gradient, simultaneously combined with eluent flow-rate gradients, very rapid simultaneous separations of both weakly retained anions and strongly retained polarisable anions was possible, with up to 10-fold decreases in overall run times. Coating stability and retention times under isocratic and isofluentic eluent conditions were shown to be reproducible over >450 repeat injections, with peak efficiency values averaging 29,000 N/m for the 2.5 cm column and 42,000 N/m for the 10 cm monolithic column, again under isocratic elution conditions.     

Evaluation of different electrolyte systems and on-line preconcentrations for the analysis of haloacetic acids by capillary zone electrophoresis

The separation of various carboxylic acids was performed on a polymethacrylate-based weakly acidic cation-exchange resin (TSKgel OApak-A) using ion-exclusion chromatography under the acidic elution conditions. When a diluted sulfuric acid solution was used as the eluent, highly sensitive conductimetric detection of carboxylic acids was achieved without increasing the background conductance of the eluent. This method was more sensitive than using benzoic acid eluent and enabled a good resolution of dicarboxylic as well as monocarboxylic acids. The addition of 5–20% methanol to the eluent considerably reduced the retention times of carboxylic acids with hydrophobic nature.