intro to chromatography

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  • intro to chromatography

    1. 1. INTRODUCTION TOCHROMATOGRAPHYPusat Pengajian Sains Kimia & Teknologi Makanan, 1Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    2. 2. What is Chromatography? Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    3. 3. Solvent extraction: Extraction is the transfer of a solute from one phase to another Phase 2 Phase 1 S(in phase 1) S(in phase 2) Extraction of Uranyl Nitrate from water into [S]2 ether Partition 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. 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 flow Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    5. 5. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    6. 6. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    7. 7. What is Chromatography? Air flowPusat Pengajian Sains Kimia & Teknologi Makanan,Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    8. 8. What is Chromatography? Fresh solvent (eluent) Initial band A and B solutes A Column packing (stationary phase) BSuspended in solvent (mobile phase) Solvent flowing out A emerges (eluate) B emerges Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    9. 9. Pusat Pengajian Sains Kimia & Teknologi Makanan, 9Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    10. 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 time Mobile phase: Stationary phase: solvent moving through substance which is fixed in the plate or column place for the chromatographic (liquid or gas) procedure The partitioning of solutes between the mobile and stationary phases gives rise to separation All chromatographic separations depend on the reversiblesorption and desorption of the components of the mixture in the stationary and mobile phase Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    11. 11. Chromatographic principle Mobile phase Equilibrium Sample established mixture at each point (ideally) Stationary phaseThe molecules of the Retardation ofmixture interact with rate ofthe molecules of the movement ofMobile and Stationary molecules Phase Different distribution Each molecule interacts coefficients and differentdifferently with MP and SP net rates of migration Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    12. 12. Purpose of ChromatographyAnalytical Chromatography – separate andidentify or measure the relative proportions ofcomponents in a mixture 300 LPreparative Chromatography - purify andcollect one or more components of a sample forfurther useThe two are not mutually exclusiveScaling Up: 2 Large mass Large column radius = Small mass Small column radius Purifies 1 Kg of material Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    13. 13. Types of Chromatography Chromatography is divided intocategories on the basis of the mechanism of interaction of the solute with the stationary phase Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    14. 14. Pusat Pengajian Sains Kimia & Teknologi Makanan, 14Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    15. 15. Pusat Pengajian Sains Kimia & Teknologi Makanan, 15Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    16. 16. Pusat Pengajian Sains Kimia & Teknologi Makanan, 16Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    17. 17. Pusat Pengajian Sains Kimia & Teknologi Makanan, 17Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    18. 18. Pusat Pengajian Sains Kimia & Teknologi Makanan, 18Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    19. 19. Pusat Pengajian Sains Kimia & Teknologi Makanan, 19Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    20. 20. Pusat Pengajian Sains Kimia & Teknologi Makanan, 20Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    21. 21. Classification based on the chromatographic principle Stationary Type of Mobile Phase Stationary Phase Phase Support Chromatography Acronym Gas Column Gas-Solid Chromatography GC/GSC Adsorption Liquid column Chromatography Chromatography LC Column High Performance LiquidCompetition between a solid HPLC Liquid Chromatography adsorbent and the mobile phase Thin Layer Chromatography TLC Planar layer Paper Chromatography PC Gas-Liquid Chromatography Partition Gas Column Supercritical Fluid GC/GLC SFC Chromatography ChromatographyCompetition between a liquid Liquid-Liquid Chromatography stationary phase and the Liquid Column High Performance Liquid LC/HPLC mobile phase Chromatography Ion Exchange Chromatography Ion-Exchange Chromatography IECCompetition between an ionic Liquid Column High Performance Ion IC/HPIC exchange resin stationary Chromatographyphase and liquid mobile phase Permeation Chromatography Competition between a Liquid Column Gel Permeation Chromatography GPC polymer matrix stationaryphase and liquidPusat Pengajian Sains Kimia & Teknologi Makanan, mobile phase Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    22. 22. Classification of Chromatography Based 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. 23. Classification according to the phases Chromatography Mobile Phase = Liquid Mobile Phase = Gas Liquid Chromatography Gas Chromatography Stationary Stationary Stationary Stationary Phase = Solid Phase = Liquid Phase = Solid Phase = Liquid LSC LLC GSC GLC Partition of Partition ofAdsorption or ion analytes analytes Adsorption exchange between two between two phases phases Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    24. 24. Classification according to physical features of the analytes Molecular Physical property Separation technique characteristic volatility GLC Polarity solubility LLC adsorptivity LSC Ion Exchange Chrom. Ionic charge Electrophoresis Gel Permeation Chrom. diffusion Size (mass) Dialysis sedimentation Ultracentrifugation Shape ligand binding Affinity Chrom. Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    25. 25. Methods based on Ionic Nature of the sampleIon nature ofthe sample is •Number & nature of ionisable groups affected by: •pH environment •Presence of other ions And results in: Association of ions Movement in an electric with opposite charge field Ion-exchange Electrophoresis chromatography Capillary electrophoresis Direction & velocity of Competition for ionic movement depends on binding sites on a resin the sign & intensity of the ionic charge Pusat Pengajian Sains Kimia & Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    26. 26. Methods based on polarity (i.e. affinity of like-molecules for each other)Partition between two phases: Solid/Liquid Liquid/Liquid Liquid/VapourA major factor in separation is: Adsorption Solubility SolubilityAnd the methods involve: Solid Two immiscible A solution adsorbents liquids and 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. 27. Methods based on sizeSize affects the rate of: Diffusion Sedimentation Movement due to this may be: Restricted by Balanced by Reduced by high Maximal in low pore size each other density solvents density solvents Isodensity Velocity DiffusionFixed pore Variable pore Ultracentrifugationsize size Dialysis Gel 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. 28. Pusat Pengajian Sains Kimia & Teknologi Makanan, 28Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    29. 29. Pusat Pengajian Sains Kimia & Teknologi Makanan, 29Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    30. 30. Pusat Pengajian Sains Kimia & Teknologi Makanan, 30Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    31. 31. Pusat Pengajian Sains Kimia & Teknologi Makanan, 31Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    32. 32. Pusat Pengajian Sains Kimia & Teknologi Makanan, 32Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    33. 33. Pusat Pengajian Sains Kimia & Teknologi Makanan, 33Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    34. 34. Pusat Pengajian Sains Kimia & Teknologi Makanan, 34Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    35. 35. Pusat Pengajian Sains Kimia & Teknologi Makanan, 35Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    36. 36. Pusat Pengajian Sains Kimia & Teknologi Makanan, 36Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    37. 37. Pusat Pengajian Sains Kimia & Teknologi Makanan, 37Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    38. 38. Pusat Pengajian Sains Kimia & Teknologi Makanan, 38Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    39. 39. Pusat Pengajian Sains Kimia & Teknologi Makanan, 39Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    40. 40. Pusat Pengajian Sains Kimia & Teknologi Makanan, 40Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    41. 41. Pusat Pengajian Sains Kimia & Teknologi Makanan, 41Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    42. 42. Pusat Pengajian Sains Kimia & Teknologi Makanan, 42Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    43. 43. Pusat Pengajian Sains Kimia & Teknologi Makanan, 43Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    44. 44. Pusat Pengajian Sains Kimia & Teknologi Makanan, 44Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    45. 45. Pusat Pengajian Sains Kimia & Teknologi Makanan, 45Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    46. 46. Pusat Pengajian Sains Kimia & Teknologi Makanan, 46Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    47. 47. Pusat Pengajian Sains Kimia & Teknologi Makanan, 47Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    48. 48. Pusat Pengajian Sains Kimia & Teknologi Makanan, 48Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    49. 49. Pusat Pengajian Sains Kimia & Teknologi Makanan, 49Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    50. 50. Pusat Pengajian Sains Kimia & Teknologi Makanan, 50Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    51. 51. Pusat Pengajian Sains Kimia & Teknologi Makanan, 51Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    52. 52. Pusat Pengajian Sains Kimia & Teknologi Makanan, 52Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    53. 53. Pusat Pengajian Sains Kimia & Teknologi Makanan, 53Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.
    54. 54. Pusat Pengajian Sains Kimia & Teknologi Makanan, 54Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia.

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