Chromatography And Spectroscopy

1. Chromatography And Spectroscopy Presented to: Dr-IngNaveedRamzan Presented by: 2007-chem-46

2. Chromatography • Physical technique for separating mixtures into their components using partitioning characteristics of molecule between the stationary phase and mobile phase in order to analyze, identify, purify, and/or quantify the mixture or components.

3. How does chromatography works? • In all chromatography techniques sample is transported in a mobile phase. The mobile phase can be a gas, a liquid, a superficial fluid. • The mobile phase is then forced through a stationary phasein column or on a solid surface. The stationary phase needs to be something that doesn’t react with the mobile phase or sample. • The sample is then has opportunity to interact with stationary phase as it moves past it. Sample that interact greatly, then appear to move more slowly. • Sample that interact weakly, then appear to move more quickly. This difference in rates, separates sample into their components.

4. How does chromatography works?

5. Chromatogram • Graph showing detector response as a function of time.

6. Chromatogram • tr retention time = time between injection and detection of the analyte. • tm= time at which the mobile phase travels through the column. • Relative retention (separation factor) α = t’r2/t’r1 • a ratio of relative retention times α > 1, indicates quality of the separation; ↑α = greater separation • Capacity factor k = (tr-tm)/tm ↑k = greater retentionα = k2/k1 • used to monitor performance of the column

7. Qualitative and quantitative analysis • Retention time • Tell us about compound identity • Peak area • How much of the compound is there

8. Retention factor Rf • A number that represents how far a compound travels in a particular solvent • It is determined by measuring the distance the compound traveled and dividing it by the distance the solvent traveled.

9. Classification Of Chromatography • According to the physical state • According to the shape of the chromatographic bed • According to the mechanism of separation

10. According To The Physical State • Gas Chromatography • High Performance Liquid Chromatography

11. Gas chromatography

12. Liquid Chromatography

13. According To The Shape Of The Chromatographic Bed • Column Chromatography • Planar chromatography • Paper chromatography • Thin layer chromatography

14. According To The Mechanism Of Separation • Adsorption chromatography • Ion exchange chromatography

15. According To The Mechanism Of Separation • Partition chromatography

16. Uses Of Chromatography • In chemical plant to separate particles and contaminates • To separate toxic material from drinking water and for monitoring of air quality. • For preparing pure raw materials and checking contaminants from manufactured compounds. • For identification of compounds.

17. Spectroscopy • A physical technique for the study of matter and its properties by the application of with the electromagnetic radiation which is represented in form of spectrum

18. Spectroscope Receives and analyze radiant energy.

19. Types of spectroscopy

20. Spectral Properties, Application and Interactions of Electromagnetic Radiation Type Radiation Type spectroscopy Energy Wave Number V Wavelength λ Frequency υ eV cm-1 cm Hz Kcal/mol Gamma ray Gamma ray emission X-ray absorption, emission X-ray Ultra violet UV absorption Visible IR absorption Infrared Microwave absorption Micro-wave Nuclear magnetic resonance Radio

21. Quantitative Analysis • Spectra is formed by the transition of electron from higher energy level to lower energy level or vice versa. The energy difference is given by the following relation E1- E0 = h u = hC/ l • l = Wave Length • C = Velocity of Radiation (constant) = 3 x 1010 cm/sec. • u = Frequency of Radiation (cycles/sec) • h (Planck's constant) = 6.62 x 10-27 (Ergsec)

22. Lambert - Beer law • When a beam of monochromatic radiation is passed through a medium, the rate of decrease of intensity of radiation with the thickness of absorbing medium is directly proportional to the intensity of radiation and concentration of solution. • dl/dt ∞ IC • -dl/dt = klC • k = molar absorption coefficient

23. Uses Of Spectroscopy • Examining of the molecular structure. • Estimation of the energy levels of the ions and complexes in a chemical system along with their compositions. • Examining the intrinsic configuration and relative association and chemical shifts • Used as a detector in chromatography

24. References: • Jack, C.;”Encyclopedia of Chromatography”;3rd Ed; Volume 3,(40-42) [23-10-2010] • http://www.rpi.edu/dept/chem-eng/Biotech-Environ/CHROMO/be_types.htm [23-10-2010] • http://en.wikipedia.org/wiki/Chromatography [23-10-2010] • http://www.iupac.org/publications/pac/1993/pdf/6504x0819.pdf [23-10-2010] • http://www.buzzle.com/articles/chromatography-uses.html [23-10-2010] • http://www.sci.sdsu.edu/TFrey/Bio750/Chromatography.html [23-10-2010] • http://en.wikipedia.org/wiki/Gas_chromatography-mass_spectrometry [2-11-2010] • http://en.wikipedia.org/wiki/Spectroscopy [23-10-2010] • http://www.camo.com/resources/types_spectroscopy.html [23-10-2010] • Raymond, W.S.P;”liquid chromatography detector” ;2nd Ed; volume 33(181-191) [2-11-2010] • http://www.hinduonnet.com/thehindu/seta/2001/12/27/stories/2001122700230400.htm [2-11-2010] • http://www.news-medical.net/health/Spectroscopy-Types.aspx [2-11-2010] • http://www.ucl.ac.uk/~ucbcdab/enzpur/ionX.htm [2-11-2010]

25. THANK YOU