Click here to load reader

intro to chromatography

  • View
    917

  • Download
    1

Embed Size (px)

Text of intro to chromatography

  • 1.INTRODUCTION TOCHROMATOGRAPHYPusat Pengajian Sains Kimia & Teknologi Makanan, 1Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.

2. What is Chromatography?Pusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 3. Solvent extraction: Extraction is the transfer of a solute from one phase to another Phase 2 Phase 1S(in phase 1) S(in phase 2) Extraction of Uranyl Nitrate from water into[S]2 etherPartition coefficient: K [S]1Chromatography:Chromatography operates on the same principle as extraction,but one phase is held in place while the other moves past it. Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 4. What is Chromatography?Chromatography:Chromatography operates on the same principle asextraction, but one phase is held in place while the othermoves past it. Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 5. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 6. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 7. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 8. What is Chromatography? Fresh solvent(eluent) Initial band A and B solutesAColumn packing (stationary phase)BSuspended in solvent (mobile phase) Solvent flowing outA emerges(eluate)B emerges Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 9. Pusat Pengajian Sains Kimia & Teknologi Makanan, 9Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 10. What is Chromatography? It is the collective term for a family of laboratory techniques for the separation of mixtures by distribution of its components between a mobile and stationary phase over timeMobile phase:Stationary phase:solvent moving through substance which is fixed inthe plate or columnplace for the chromatographic(liquid or gas)procedureThe partitioning of solutes between the mobile andstationary phases gives rise to separation All chromatographic separations depend on the reversiblesorption and desorption of the components of the mixture inthe stationary and mobile phase Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 11. Chromatographic principleMobile phaseEquilibrium Sample established mixtureat each point(ideally)Stationary phaseThe molecules of theRetardation ofmixture interact withrate ofthe molecules of themovement ofMobile and Stationary moleculesPhase Different distribution Each molecule interacts coefficients and differentdifferently with MP and SPnet rates of migrationPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 12. Purpose of ChromatographyAnalytical Chromatography separate andidentify or measure the relative proportions ofcomponents in a mixture300 LPreparative Chromatography - purify andcollect one or more components of a sample forfurther useThe two are not mutually exclusiveScaling Up:2Large mass Large column radius = Small massSmall column radiusPurifies 1 Kg of materialPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 13. Types of Chromatography Chromatography is divided intocategories on the basis of the mechanismof interaction of the solute with thestationary phasePusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 14. Pusat Pengajian Sains Kimia & Teknologi Makanan, 14Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 15. Pusat Pengajian Sains Kimia & Teknologi Makanan, 15Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 16. Pusat Pengajian Sains Kimia & Teknologi Makanan, 16Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 17. Pusat Pengajian Sains Kimia & Teknologi Makanan, 17Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 18. Pusat Pengajian Sains Kimia & Teknologi Makanan, 18Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 19. Pusat Pengajian Sains Kimia & Teknologi Makanan, 19Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 20. Pusat Pengajian Sains Kimia & Teknologi Makanan, 20Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 21. Classification based on the chromatographic principle Stationary Type ofMobile PhaseStationary PhasePhase Support Chromatography Acronym Gas Column Gas-Solid Chromatography GC/GSCAdsorptionLiquid column ChromatographyChromatography LC ColumnHigh Performance LiquidCompetition between a solid HPLCLiquid Chromatography adsorbent and the mobilephaseThin Layer Chromatography TLC Planar layer Paper ChromatographyPC Gas-Liquid Chromatography Partition Gas ColumnSupercritical Fluid GC/GLCSFCChromatographyChromatographyCompetition between a liquidLiquid-Liquid Chromatographystationary phase and theLiquid ColumnHigh Performance Liquid LC/HPLCmobile phase Chromatography Ion ExchangeChromatographyIon-Exchange Chromatography IECCompetition between an ionicLiquid Column High Performance Ion IC/HPICexchange resin stationaryChromatographyphase and liquid mobile phasePermeationChromatography Competition between aLiquidColumn Gel Permeation Chromatography GPCpolymer matrix stationaryphase and liquidPusat Pengajian Sains Kimia & Teknologi Makanan, mobile phase Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 22. Classification of ChromatographyBased on Stationary Phase SupportPlanar: Stationary Phase supported on a flat plateor in pores of paper. Mobile Phase moves throughStationary Phase by capillary action, gravity orelectrosatic forcesColumn: Stationary Phase held in a tube andMobile Phase forced through Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 23. Classification according to the phases Chromatography Mobile Phase = LiquidMobile Phase = GasLiquid ChromatographyGas Chromatography StationaryStationary Stationary StationaryPhase = SolidPhase = LiquidPhase = Solid Phase = Liquid LSCLLCGSCGLC Partition ofPartition ofAdsorption or ion analytesanalytes Adsorption exchange between two between two phasesphasesPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 24. Classification according to physical features of theanalytesMolecular Physical propertySeparation techniquecharacteristicvolatilityGLC Polarity solubilityLLCadsorptivityLSCIon Exchange Chrom.Ionic charge Electrophoresis Gel Permeation Chrom.diffusion Size (mass)Dialysis sedimentation Ultracentrifugation Shape ligand bindingAffinity Chrom. Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 25. Methods based on Ionic Nature of the sampleIon nature ofthe sample isNumber & nature of ionisable groups affected by: pH environment Presence of other ions And results in:Association of ionsMovement in an electric with opposite charge field Ion-exchange Electrophoresischromatography Capillary electrophoresis Direction & velocity of Competition for ionic movement depends onbinding sites on a resin the sign & intensity ofthe ionic chargePusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 26. Methods based on polarity (i.e. affinity of like-molecules for each other)Partition between two phases:Solid/LiquidLiquid/Liquid Liquid/VapourA major factor in separation is:AdsorptionSolubilitySolubilityAnd the methods involve:Solid Two immiscibleA solutionadsorbents liquidsand its vapourMethods are generallyknown as: Adsorption Liquid Gas-Liquid Chromatography Chromatography Chromatography Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 27. Methods based on sizeSize affects the rate of: DiffusionSedimentationMovement due tothis may be:Restricted byBalanced by Reduced by highMaximal in lowpore sizeeach otherdensity solvents density solventsIsodensity Velocity DiffusionFixed pore Variable poreUltracentrifugationsize size DialysisGel permeation chromatography Gel filtration Molecular exclusion (largest molecules come out first as not held by pores) Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 28. Pusat Pengajian Sains Kimia & Teknologi Makanan, 28Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 29. Pusat Pengajian Sains Kimia & Teknologi Makanan, 29Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 30. Pusat Pengajian Sains Kimia & Teknologi Makanan, 30Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 31. Pusat Pengajian Sains Kimia & Teknologi Makanan, 31Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 32. Pusat Pengajian Sains Kimia & Teknologi Makanan, 32Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 33. Pusat Pengajian Sains Kimia & Teknologi Makanan, 33Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 34. Pusat Pengajian Sains Kimia & Teknologi Makanan, 34Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 35. Pusat Pengajian Sains Kimia & Teknologi Makanan, 35Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 36. Pusat Pengajian Sains Kimia & Teknologi Makanan, 36Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 37. Pusat Pengajian Sains Kimia & Teknologi Makanan, 37Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 38. Pusat Pengajian Sains Kimia & Teknologi Makanan, 38Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 39. Pusat Pengajian Sains Kimia & Teknologi Makanan, 39Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 40. Pusat Pengajian Sains Kimia & Teknologi Makanan, 40Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 41. Pusat Pengajian Sains Kimia & Teknologi Makanan, 41Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 42. Pusat Pengajian Sains Kimia & Teknologi Makanan, 42Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 43. Pusat Pengajian Sains Kimia & Teknologi Makanan, 43Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 44. Pusat Pengajian Sains Kimia & Teknologi Makanan, 44Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 45. Pusat Pengajian Sains Kimia & Teknologi Makanan, 45Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 46. Pusat Pengajian Sains Kimia & Teknologi Makanan, 46Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 47. Pusat Pengajian Sains Kimia & Teknologi Makanan, 47Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 48. Pusat Pengajian Sains Kimia & Teknologi Makanan, 48Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 49. Pusat Pengajian Sains Kimia & Teknologi Makanan, 49Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 50. Pusat Pengajian Sains Kimia & Teknologi Makanan, 50Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 51. Pusat Pengajian Sains Kimia & Teknologi Makanan, 51Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 52. Pusat Pengajian Sains Kimia & Teknologi Makanan, 52Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 53. Pusat Pengajian Sains Kimia & Teknologi Makanan, 53Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia. 54. Pusat Pengajian Sains Kimia & Teknologi Makanan, 54Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.