High-effective IONIC CHROMATOGRAPHY
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This document discusses high-effective ionic chromatography. It defines it as a process that separates ionic compounds based on their charge and affinity for ion exchangers. There are two main types: cation exchange chromatography which uses a negatively charged stationary phase, and anion exchange chromatography which uses a positively charged stationary phase. The instrumentation includes a pump, injector, column, suppressor, detector and recorder. Samples are separated as they pass through the column based on differences in how strongly ions bind to it. Applications include water softening, demineralization, and separating substances like amino acids, proteins, ions and pharmaceutical compounds.
![References:
[1] Ion Exchange Chromatography & Chromatofocusing - Principles and Methods, “Ion Exchange
Chromatography & Chromatofocusing - Principles and Methods,” GE Healthc., pp. 1–188,
2004.
[2] GE Healthcare, “Ion Exchange Columns,” pp. 1–8, 2011.
[3] H. F. Walton, “Ion Exchange Chromatography,” Anal. Chem., vol. 36, no. 5, pp. 51–55, 1964,
doi: 10.1021/ac60211a004.
[4] G. E. Healthcare, “Ion Exchange Chromatography & Chromatofocusing: Principles and
Methods,” GE Heal. Handbooks, p. 170, 2016.
[5] O. Bobleter and G. Bonn, “Ion exchange chromatography,” Ion Exch., no. May, pp. 1187–
1242, 2011, doi: 10.1515/9783110862430.1187.
[6] &NA;, “Learning Objectives,” Contin. Lifelong Learn. Neurol., vol. 20, p. 519, 2014, doi:
10.1212/01.con.0000450959.48636.df.
[7] H. Ullah, “The Role of Ion Exchange Chromatography in Purification and Characterization of
Molecules,” Ion Exch. Technol., 2012, doi: 10.5772/52537.
[8] P. Stanton, “3,” vol. 251.
[9] M. Jaworski, “Ion-exchange chromatography of the product obtained in a reaction of
ethylene diamine with urea,” Chromatographia, vol. 13, no. 2, pp. 101–104, 1980, doi:
10.1007/BF02263061.
[10] S. Together, “Separation of proteins with anion exchange chromatography on Sepapure Q
and DEAE Separation of proteins with anion exchange chromatography on Sepapure Q and
DEAE,” pp. 1–4.
[11] P. Of and I. O. N. Exchange, “Instrumentation PRINIPLES OF ION EXCHANGE.”
[12] Mohammed and JS, “A brief review on Ion Exchange Chromatography,” PharmaTutor, vol. 5,
no. 2, pp. 30–38, 2017, [Online]. Available: https://www.pharmatutor.org/articles/breif-
review-on-ion-exchange-chromatography.
[13] O. Bahadir, “Ion-Exchange Chromatography and Its Applications,” Column Chromatogr.,
2013, doi: 10.5772/55744.](https://image.slidesharecdn.com/ipec-210623092250/85/High-effective-IONIC-CHROMATOGRAPHY-26-320.jpg)
High-effective IONIC CHROMATOGRAPHY
- 1. High-effective IONIC CHROMATOGRAPHY Eng. Mustafa Ahmed Khaldoun Termanini
- 2. The main titles: Definition of High-effective Ionic Chromatography. Separation principle. Types of High-effective Ionic Chromatography. Instrumentation of High-effective Ionic Chromatography. Steps of the process of using High-effective Ionic Chromatography. Applications of High-effective Ionic Chromatography.
- 3. Definition of High-effective Ionic Chromatography - It is an analytical technical process by which ionic compounds are separated based on their charge and affinity with ion exchangers. Separation principle: The reverse exchange between the ionic compounds present in the mobile phase (liquid) that contains the sample and the stationary phase (solid) which contains insoluble substances and ionic sites.
- 4. TYPES OF High-effective IONIC CHROMATOGRAPHY
- 5. A. Cation exchange chromatography: - in it, the stationary phase (solid supports) have negatively charged functional groups It is used to separate molecules that have a positive charge. R-X−𝐂+ + 𝐌+B− ⇄ R-X−𝐌+ + C+ +B− - Commonly used cation exchange resins are S-resin, sulfate derivatives, and CM resins, carboxylate derived ions.
- 6. B. Anion exchange chromatography: - in it, the stationary phase (solid supports) have positively charged functional group It is used to separate molecules that have a negative charge. R-X+𝐀− + M+𝐁− ⇄ R-X+𝐁− + M+ + 𝐀− - Commonly used anion exchange resins are Q-resin, a Quaternary amine; and DEAE resin, Diethyl Aminoethane.
- 7. Instrumentation of High-effective Ionic Chromatography Instrumentation includes: 1. Pump. 2. Injector. 3. Column. 4. Suppressor. 5. Oven(optional). 6. Detector. 7. Recorder or data system.
- 8. 1. PUMP: • Is one of the most important components in the system. • Which must provide a continuous constant flow of the eluent through the injector, column, and detector. • A high-pressure pump with pressure and flow indicator, to deliver the eluent. • The pressure required for most instruments is at least 600 psi up to 4000 psi.
- 9. 2. INJECTOR: - The sample is manually injected into the column by injection valve using a syringe. - Or automatically by using an automatic sampler.
- 10. 3. COLUMN: • Used to separate the sample mixture into the individual components. • its diameter from about 2mm to 5 cm and in length from 3 cm to 50cm. • is made of glass, stainless steel, titanium, or polymers.
- 11. The suppressor reduces the background conductivity of the chemicals used to elute samples from the ion-exchange column which improves the conductivity measurement of the ions being tested and enhancing the sensitivity. 4. OVEN (optional). 5. SUPPRESSOR:
- 12. 6. DETECTORS: Detection methods include: • Mass spectrometry. • Atomic spectroscopy. • Fluorescence. • Luminescence. • UV-vis, potentiometric. • Electrical conductivity detector is commonly used. - Conductivity detector is the measure of a material’s ability to conduct electricity. Since conductivity is proportional to the number of ions in solution, it is the primary method of detection for ion chromatography.
- 13. 7. RECORDER OR DATA SYSTEM: - For collecting and organizing the chromatograms and data such as a data station or minicomputer.
- 14. Steps of the process of using High-effective Ionic Chromatography:
- 15. Steps of the process of using High-effective Ionic Chromatography: 1. Eluent Loading 2. Sample injection 3. Separation of sample 4. Elution of analyte A 5. Elution of analyte B : Eluent ion . : Ion A . : Ion B .
- 16. Steps of the process of using High-effective Ionic Chromatography: 1. Eluent Loading: The eluent loaded onto the column displaces any anions bonded to the resin and saturates the resin surface with the eluent anion. 2. Sample injection: A sample containing anion A and anion B are injected in the column. This sample could contain many different ions. : Eluent ion . : Ion A . : Ion B .
- 17. Steps of the process of using High-effective Ionic Chromatography: 3. Separation of sample: After the sample has been injected, the continued addition of eluent causes a flow through the column. As the sample elutes (or moves through the column), anion A and anion B adhere to the column surface differently. The sample zones move through the column as eluent gradually displaces the analytes. : Eluent ion . : Ion A . : Ion B .
- 18. Steps of the process of using High-effective Ionic Chromatography: 4. Elution of analyte A: As the eluent continues to be added, the anion A moves through the column in a band and ultimately is eluted first. 5. Elution of analyte B: The eluent displaces anion B, and anion B is eluted off the column. : Eluent ion . : Ion A . : Ion B .
- 19. Stationary phase: usually made from inert organic matrix chemically derivative with ionizable functional groups (fixed ions) which carry displaceable oppositely charged ion. The main classes of substances used are: • Inorganic salts , Zeolites. • Metal oxides and cellulose. • Modified organic polymer resins, modified silica gels, (The most used). Mobile phase: Acids, alkalis, Buffers. Such as: HCl, NaOH, phosphate buffers, acetate buffers, borate buffers, phthalate buffers.
- 20. RESINS - Are made of polystyrene sulfonate. CLASSIFICATION OF RESINS: According to the chemical nature they classified as: 1. Strong cation exchange resin – Sulphonic acid. 2. Weak cation exchange resin – Carboxy methyl compound. 3. Strong anion exchange resin – Quaternary ammonium compound. 4. Weak anion exchange resin - Diethyl aminoethyl compound. Regeneration of ion exchange resin: The procedure is performed using strong acids and alkalis such as hydrochloric acid and sodium hydroxide
- 21. Applications of High-effective Ionic Chromatography
- 22. Applications of High-effective Ionic Chromatography: 1. Softening of hard water: The hardness of water is due to the presence of 𝐂𝐚+𝟐, 𝐌𝐠+𝟐, and other divalent ions that may be removed by passing the hard water through the cation exchanger charged with 𝐍𝐚+ ions. 2. Complete demineralization of water: The ions (cations and anions) are completely removed by passing water through two cation and anion exchangers.
- 23. Applications of Ion High-effective Ionic Chromatography: 3. Separation of amino acids and protein and small nucleotides: Each amino acids and protein have Isoelectric Point PI: It is a point at which protein charges are equal to those of the surrounding medium, so that proteins precipitate, for example casein precipitates at a pH of 4.6, and is used in the separation of proteins and amino acids.
- 24. Applications of High-effective Ionic Chromatography: 4. Separation of different ions. 5. Other applications: • For the measurement of various active ingredients in medicinal formulations. • For the measurement of drugs and their metabolites in serum and urine, for residue analysis in food raw materials. • For the measurement of additives such as vitamins and in food and beverages.
- 25. References
- 26. References: [1] Ion Exchange Chromatography & Chromatofocusing - Principles and Methods, “Ion Exchange Chromatography & Chromatofocusing - Principles and Methods,” GE Healthc., pp. 1–188, 2004. [2] GE Healthcare, “Ion Exchange Columns,” pp. 1–8, 2011. [3] H. F. Walton, “Ion Exchange Chromatography,” Anal. Chem., vol. 36, no. 5, pp. 51–55, 1964, doi: 10.1021/ac60211a004. [4] G. E. Healthcare, “Ion Exchange Chromatography & Chromatofocusing: Principles and Methods,” GE Heal. Handbooks, p. 170, 2016. [5] O. Bobleter and G. Bonn, “Ion exchange chromatography,” Ion Exch., no. May, pp. 1187– 1242, 2011, doi: 10.1515/9783110862430.1187. [6] &NA;, “Learning Objectives,” Contin. Lifelong Learn. Neurol., vol. 20, p. 519, 2014, doi: 10.1212/01.con.0000450959.48636.df. [7] H. Ullah, “The Role of Ion Exchange Chromatography in Purification and Characterization of Molecules,” Ion Exch. Technol., 2012, doi: 10.5772/52537. [8] P. Stanton, “3,” vol. 251. [9] M. Jaworski, “Ion-exchange chromatography of the product obtained in a reaction of ethylene diamine with urea,” Chromatographia, vol. 13, no. 2, pp. 101–104, 1980, doi: 10.1007/BF02263061. [10] S. Together, “Separation of proteins with anion exchange chromatography on Sepapure Q and DEAE Separation of proteins with anion exchange chromatography on Sepapure Q and DEAE,” pp. 1–4. [11] P. Of and I. O. N. Exchange, “Instrumentation PRINIPLES OF ION EXCHANGE.” [12] Mohammed and JS, “A brief review on Ion Exchange Chromatography,” PharmaTutor, vol. 5, no. 2, pp. 30–38, 2017, [Online]. Available: https://www.pharmatutor.org/articles/breif- review-on-ion-exchange-chromatography. [13] O. Bahadir, “Ion-Exchange Chromatography and Its Applications,” Column Chromatogr., 2013, doi: 10.5772/55744.
- 27. THANK YOU Eng. Mustafa Ahmed Khaldoun Termanini