ion exclusion chromatography.pptx

ion exclusion chromatography
M. Pharma
pharmaceutical chemistry
2nd sem

1. CHROMATOGRAPHY- ION-EXCLUSION CHROMATOGRAPHY Department of Pharmaceutical Sciences Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur - 440033 Presented by- Tahmina P. Y. Khan M.Pharm . 1 ST Year (Pharmaceutical Chemistry) Semester-II 1

2. Content Introduction Principle Instrumentation Application References 2

3. Introduction First introduced by Wheaton and Bauman in 1953. ION-EXCLUSION CHROMATOGRAPHY is a technique used to separate ionic compounds from non-ionic compounds and to separate mixtures of acids. Useful technique for the separation of ionic and nonionic substances. It involves the use of strong anion- or cation- exchange resins for the separation of ionic solutes from weakly ionized or neutral solutes. 3

4. Principle Ionic substances pass quickly through the column. Non-ionic (molecular) or partially ionized substances are held up and are eluted more slowly. Functions by Donnan exclusion mechanism. Mobile phase- liquid (water or aqueous acid or base) Stationary phase- ion exchange resins; Cationic Anionic 4

5. Chromatographic system consists of 3 phases. 5

6. Schematic Illustration Of Ion Exclusion- 6

7. Thus, analytes are separated by: Exclusion or repulsion if they are ionized and have the same charge as the resin. Adsorption if they are non-polar or partially ionized. 7

8. Instrumentation Stationary Phase 8

9. Application of the sample- After packing sample is added to the top of the column, use syringe or pipette. Mobile phase- Acid, alkali, buffers Iv. Elution- Components of mixture separated & move down the column at different rates depending upon the affinity of the ion for ion exchanger. Analysis of the eluate- Spectrophotometric Conductivity. 9

10. Schematic Illustration Of Instrumentation ION-EXCLUSION CHROMATOGRAPHY 10

11. ELUENTS: 3 types of eluents are used in ion exclusion chromatography- Water eluent Acid eluent - dilute solutions of strong mineral acids ,or dilute solutions of strong bases. Complexing eluent - An example of this approach is the use of a mannitol eluent for the determination of boric acid, in which the mannitol serves to complex the boric acid to form a species which is more easily detectable by conductivity measurements than is boric acid alone). 11

12. DEGASSING UNIT ANALYTICAL COLUMN- The analytical column consists of stationary phase (wet packing) The column packing consists of a reactive layer bonded to inert polymeric particles. SUPPRESSOR- Suppression plays a key role in the analysis of anions and organic acids using ion chromatography and conductivity detection. For anion analysis, the suppressor is a high-capacity cation exchange membrane or resin in the acid form. It removes cations from the eluent and replaces them with H+. 12

13. Detector: - Conductivity Detector: - In principle, the conductivity detector can be used for some non-aqueous eluents. The sensitivity of these detectors depends on the temperature during the separation and detection the temperature must be kept strictly constant. 13

14. Fluorescent based Detectors Detectors with UV- Visible spectrophotometry 14

15. Application Determination of neutral molecules: - Neutral compounds such as sugars and alcohols can be separated by IEC, Determination of strong inorganic acids: - Sulphate, nitrate and chloride ions, and strong base cations such as sodium, ammonium, potassium. Magnesium and calcium ions commonly found in acid rainwater. Separation of carboxylic acids: - The separation of carboxylic acids is the most common application of ion-exclusion chromatography. 15

16. Determination of Weak inorganic acids and bases: - IEC has found increasing use for the determination of weakly ionized inorganic anions such as fluoride, nitrite, phosphate and cyanide. This approach is very effective for the determination of weakly ionized anions in samples containing a high concentration of ionic species, e.g., seawater and wastewaters. Determination of water in some organic solvents. Determination of amino acids and its derivatives. 16

17. References M. Mori, K. Tanaka, Q. Xu, M. Ikedu , H. Taoda , W. Hu, "Highly sensitive determination of hydrazine ion by ion-exclusion chromatography with ion- exchange enhancement of conductivity detection." Journal of Chromatography A 1039, no. 1-2 (2004): 135-139. B. K. Glód , "Ion exclusion chromatography: parameters influencing retention." Neurochemical research 22 (1997), page no: 1237-1248 17

18. THANK YOU. 18