Acceptance of ion chromatography for anion analysis was very rapid, mainly due to the lack of alternative methods that could quickly and accurately determine several anions in a single analysis. The situation regarding analysis of cations was quite different.
The total ionic composition is known using a single analytical technique. Modern ion chromatography was first reported in in a landmark paper by Small, Stevens, and Bauman.
In September ion chromatography was publicly presented at a meeting of the American Chemical Society, where the first commercially available instrument for performing ion chromatography was exhibited. Several years later Gjerde et al developed a variety of ion chromatography, a non-suppressed ion chromatography technique.
At present there are two main types of ion chromatography: suppressed ion chromatography and non-suppressed ion chromatography. Ion exchange remains the primary separation mode used in ion chromatography today, although the apparatus used for the separation of the ionic species includes ion pairing, ion exclusion and chelation chromatography.
Ion chromatography can be used for the determination of ionic solutes such as inorganic anions, inorganic cations including alkali metals, alkaline earth metals, transition metals, and rare earth metals , carboxylic, phosphonic and sulfonic acids, detergents, carbohydrates, low molecular weight organic bases, and ionic metal complexes.
Until only a small range of analytical parameters could be measured automatically. It was necessary therefore to develop and validate new methods to extend the scope of such parameters. An alternative that has almost replaced most of the wet chemical methods used in water analysis, is ion chromatography. For laboratories that need to determine numerous anions and cations in several thousand samples, ion chromatography is an attractive technique. It is especially attractive when the laboratories do not have the throughput to justify the purchase of large automatic analysers, which are usually based on colorimetric procedures.
Ion chromatography eliminates the need to use the hazardous reagents that are often integral to wet chemical methods. The separation and detection methods employed in ion chromatography, as well as the selection of ionic species that may be analysed with the ion chromatography method, are summarised in table one and table two. A survey of the detection methods used is given in table three.
Regulators and clients expect to receive accurate and comparable results from a laboratory. Legislators generally define which of various validated methods should be applied to analyse selected samples. In general, standard methods can be chosen to serve as reference methods; however, the laboratory serving a public client should apply official reference methods.
Ion chromatography methods meet these requirements and can be used for routine applications in laboratories. Standardisation on an international level is the responsibility of the International Standardization Organization ISO. Standard methods can be adopted as recommended on a voluntary basis by any laboratory around the world.
In addition, governments can decide to incorporate existing standards into their national standards. After the publication of US Environmental Protection Authority EPA and particularly ISO standards concerning ion chromatography, the number of laboratories applying this technique has increased dramatically. For those laboratories, ion chromatography is a reliable and economical supplement to existing wet chemical methods. More than 20 years ago, capillary electrophoresis appeared as a promising substitute for ion chromatography, mainly because of its higher speed of separation.
Comparison of ion chromatography and capillary electrophoresis has shown that these techniques can be considered as being complementary rather than competitive. Until now there were no international standards applied for capillary electrophoretic methods. Application of ion chromatography for the determination of inorganic and organic ions mostly concerns the following areas: environmental analysis, power plant chemistry, semiconductor industry, metal processing, pharmaceutics, biotechnology, mining, agriculture, food and beverages, electroplating, and the pulp and paper industry.
Environmental analysis is one of the most important fields of application for ion chromatography: it can be divided into water, soil and air analysis.
The main focus of the application of ion chromatography in environmental analytical chemistry is the qualitative and quantitative analysis of anions and cations in all kinds of waters. Ion chromatography can be also used for the analysis of ions in natural brine waters, which include seawaters, subsurface brines, geothermal brines, and high salinity ground waters.
The determination of cyanide in various samples is very important environmentally because of its large scale industrial uses and its extreme toxicity. In all of the analytical methods developed so far for cyanide and sulfide, a necessary first step is removal of interferences. With the ion chromatography method, cyanide and sulfide are separated and are thus determined simultaneously.
Polyphosphates are widely used in industrial water treatment applications on account of their sequestering and dispersing properties. Chelating agents such as nitriltriacetate acid NTA and ethylenediaminetetraacetic acid EDTA can also be determined rapidly by the same approach used for polyphosphonates. Besides the common inorganic anions and cations, chromate and arsenite are of primary concern because of their greater toxicities as compared to chromium III and arsenate respectively.
Hexavalent chromium is a toxic form of chromium that must be monitored in manufacturing wastes. Ion chromatography with postcolumn addition of diphenylcarbazide is probably one of the most specific and sensitive methods available for the determination of hexavalent chromium.
Water treatment by disinfection processes is considered a major public health achievement of the Twentieth Century. Consequently, there has been a shift in the identification methods of water contaminants from microbiological to chemical. Table 1. Weak and Strong type anion and cation exchangers. Mobile phase Eluent In ion exchange chromatography generally eluents which consist of an aqueous solution of a suitable salt or mixtures of salts with a small percentage of an organic solvent are used in which most of the ionic compounds are dissolved better than in others in.
Commonly used eluent additives which have been successfully used in ion exchange chromatography can be given as follow; EDTA; Ethylenediamine tetraacetic acid Polyols; Glycerol, glucose, and saccharose Detergents; Urea and guanidinium chloride Lipids Organic solvents Zwitterions Sulfhydryl reagents Ligands Protease inhibitors [ 14 ].
Buffer In ion exchange chromatography, pH value is an important parameter for separation and can be controlled and adjusted carefully by means of buffer substances [ 18 ].
Substance pK a Working pH Citric acid 3. Table 2. Commonly used buffers for cation-exchange chromatography. Table 3. Commonly used buffers for anion-exchange chromatography. Detection Conductivity detector is the most common and useful detector in ion exchange chromatography. Sample 1: Source: Nigella sativa Linn. Extraction procedure: Water extract of N. Powder was dissolved in phosphate buffer saline pH 6.
The supernatant was collected as the soluble extract by removing the oily layer and unsoluble pellet. Protein concentration of the soluble extract was determined with Bradford method. Then proteins dialyzed against 0. Eluent: 0. Fractions of each were collected with an increasing concentration of NaCl Detection: UV detector at nm Analyte s : Number of protein bands ranging from kDa molecular mass [19].
Sample 2: Source: Olea europea L. Extraction procedure: Extract was prepared from the leaves and roots of two years old olive plants with water at room temperature. Internal standard as D O- methylglucopyranose MeGlu was used and added in appropriate volume. Extraction was accomplished by shaking for 15 min and finally the suspension was centrifuged at rpm for 10 min.
Before the injection the aqueous phase was filtered and passed on a cartridge OnGuard A Dionex to remove anion contaminants.
Detection: Pulsed amperometric detection Analyte s : myo -inositol, galactinol, mannitol, galactose, glucose, fructose, sucrose, raffinose and stachyose [20]. Sample 3: Source: Soybean Extraction procedure: Soybeans were defatted with petroleum ether for 30 min and centrifuged repeating the procedure twice.
Then proteins were extracted with 0. The supernatant was adjusted to pH 6. The precipitate was dissolved in Tris-HCl buffer and the process was repeated in order to obtain purified precipitated fraction containing the 11S globulin.
The supernatant obtained after the first precipitation of the 11S fraction was adjusted to pH 4. The supernatant was stored at low temperature and the precipitate was dissolved in Tris-HCl buffer pH 8.
The process was repeated to obtain a purified precipitated fraction containing the 7S globulin. In all cases, the buffer concentration was 20 mM. For every buffer, different gradients were tried. Sample 4: Source: Cochlospermum tinctorium A. Extraction procedure: The powdered roots of C. Extraction procedures continue until no color could be observed in the ethanol.
The acidic fractions were obtained by elution of linear NaCl gradient The carbohydrate elution profile was determined using the phenol-sulphiric acid method. Finally two column volumes of a 2 M sodium chloride solution in water were eluted to obtain the most acidic polysaccharide fraction. The relevant fractions based on the carbohydrate profile were collected, dialysed and lyophilized.
Detection: UV detector, nm Analyte s : Glucose, galactose, arabinose in neutral fraction Uronic acids Both galacturonic and glucuronic acid , rhamnose, galactose, arabinose and glucose in acidic fraction [22]. Sample 5: Source: Hen egg Extraction procedure: Fresh eggs were collected and the same day extract was obtained.
Ovomucin was obtained using isoelectric precipitation of egg white in the presence of mM NaCl solution. After centrifugation at The supernatants obtained during the first step with mM NaCl solution and the second step with mM NaCl solution was further used for ion exchange chromatography to separate other egg white proteins.
Separation proteins from mM supernatant were allowed to pass through an anion exchange chromatographic column to separate different fractions. The unbound fractions were then passed through a cation exchange chromatographic column to separate further. Finally the bound fraction was eluted using gradient elution 0. The unbound fraction was collected and used as starting material for cation exchange chromatography.
The column was equilibrated with 10 mM citrate buffer, which was used as the starting buffer. After sample injection the column was eluted by isocratic elution using 0. The fractions were collected and freeze dried-Cation Exchange Chromatography.
The precipitate was removed by centrifugation and the supernatant was extensively dialysed against distiled water. The dialysed protein extract was freeze dried and used for chromatographic separation. Elution of the bound fraction was carried out by using 1 M NaCl in the equilibration buffer.
Sample 7: Source: Sweet dairy whey Extraction procedure: After the cheese making process the sweet whey is produced, it is further processed by reverse osmosis to increase the solids content from approximately 5. Stationary phase: Pharmacia's Q- and S-Sepharose anion- and cation-exchange resins Eluent 1: For the anion-exchange process; it was found that two step changes, simultaneous in pH and salt concentration were necessary to carry out the anion-exchange separation.
After the whey feed was loaded onto the column, one column volume of this buffer was passed through to wash out any material that did not bind to the resin, including the IgG.
Next, two column volumes of 0. This was then followed by two column volumes of 0. After this second step change, the cleaning cycle was then implemented to prepare the column for the next run. Eluent 2: For the cation-exchange process, it was found that one step change in pH was appropriate to carry out the cation-exchange separation. The buffer used was 0.
One column volume loading of the anion-exchange breakthrough curve fraction was optimum for loading onto the cation-exchange column. After the anion-exchange breakthrough curve fraction was loaded onto the column, one column volume of the initial buffer was passed through to wash out any material that did not bind to the resin.
Next a step change in pH was implemented to elute the bound IgG. This was accomplished by passing two column volumes of the buffer, 0.
As the pH wave of this buffer passed through the cation bed it initiated the elution of the IgG because the upper value of its p I range is 8.
After this pH step change the cleaning cycle was then implemented. The buffer used was 3 ml g -1 of the fresh leaves. An aliquot of the dialysed ammonium sulfate fraction containing protein was applied to the affinity chromatography on the N -acetylgalactosamine-agarose column. And then further separation was performed on Sephacryl S column followed by anion exchange chromatography.
Extraction: Fruits of the plant extracted with hot water yielded a crude polysaccharide sample, CLRP. The carbohydrate of CLRP was CLRP was a black Polysaccharide sample in which the pigment could not be removed by colum chromatography.
After decoloration, the carbohydrate content of decolored CLRP was Crude polysaccharide material was dissolved in mL 0. Sample Source: Physalisalkekengi var.
The precipitate was dissolved in distilled water and the solution was then washed with sevag reagent isoamyl alcohol and chloroform in ratio , which were centrifuged at rpm for 15 min and the protein was removed. Total sugars were determined by the phenol—sulfuric acid assay using glucose as standard.
Stationary Phase: DEAE anion-exchange column Eluent: The column was eluted first with distilled water, and then with gradient solutions 0. The column was eluted with 0. The major fraction was collected and then freeze dried. All of these fractions were assayed for sugar content by the phenol—sulfuric acid method using glucose as standard Detection: UV Detector, nm Analyte s : Polysaccharides [29].
Sample Source: Ornithogalum caudatum Ait. The polysaccharide pellets were obtained by centrifugation at rpm for 15 min, and completely dissolved in appropriate volume of distilled water followed by intensive dialysis for 2 days against distilled water cut-off M w Da. The retentate portion was then concentrated, and centrifuged to remove insoluble material.
Finally the supernatant was lyophilized to give crude extract. The crude extract was dissolved in 0. The solution was passed through an anion-exchange chromatography column. After ion exchange chromatography other chromatographic methods was used for further separations. Detection: UV Detector, nm phenol—sulfuric acid method Analyte s : Water soluble polysaccharides [30]. The separated proteins were then re-suspended in a minimum amount of distilled water and the solution dialyzed using cellulose dialysis tubing for 24 hrs against distilled water and concentrated by freeze-drying.
The partially purified enzyme was dissolved in acetate buffer 20 mM - pH 6. The solution was passed through the column at a flow rate of 1 mL. The eluted fractions were collected in an automated fraction collector Pharmacia Biotech and the absorbance of the fractions was measured at nm. The major peak fractions were then assayed for tannase activity, and only the fractions possessing tannase activity were pooled.
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More statistics for editors and authors Login to your personal dashboard for more detailed statistics on your publications. Access personal reporting. More About Us. Exchange Type. Ion exchange group. Buffer counter ions. Commercial samples. Strong cation. Sulfonic acid SP. Weak cation. Carboxylic acid. CM Cellulose. Strong anion. Quaternary amine Q. Weak anion. DEAE Cellulose. Working pH. N - 2-acetamido iminodiacetic acid. N,N-bis 2-hydroxyethyl glycine. Bis-Tris propane. N -Methyl-diethanolamine.
Sample Nigella sativa Linn. Extraction procedure:. Water extract of N. Stationary Phase:. Fractions of each were collected with an increasing concentration of NaCl. UV detector at nm. Analyte s :. Number of protein bands ranging from kDa molecular mass [19].
Olea europea L. Extract was prepared from the leaves and roots of two years old olive plants with water at room temperature. Pulsed amperometric detection. Soybeans were defatted with petroleum ether for 30 min and centrifuged repeating the procedure twice. Cochlospermum tinctorium A.
The powdered roots of C. For obtaining neutral fraction the column was eluted with water firstly. UV detector, nm. Glucose, galactose, arabinose in neutral fraction Uronic acids Both galacturonic and glucuronic acid , rhamnose, galactose, arabinose and glucose in acidic fraction [22]. Fresh eggs were collected and the same day extract was obtained. The column was equilibrated with water and the pH was adjusted to 8.
Ovalbumin, ovotransferrin, lysozyme, ovomucin [23]. Q-Sepharose Column 3 cm x 7 cm , anion-exchange. A Da antifungal peptide [24]. After the cheese making process the sweet whey is produced, it is further processed by reverse osmosis to increase the solids content from approximately 5. Stationary phase:. Pharmacia's Q- and S-Sepharose anion- and cation-exchange resins. Eluent IC with suppressed conductivity is often the method of choice. Health Canada : Established a maximum level of benzoic acid of 1, ppm in fruit juice, jams, and packaged fish and meat products.
FDA : Established a maximum level of acetic acid of 0. CMD SchemaApp code. Sign in. Account Check Order Status. Oxalic acid Propionic acid Tartaric acid Valeric acid.
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