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Raman Spectroscopy Anatasia Gangadin Dr. Ronald Birke City College of New York What is Spectroscopy? The study of how 'species' (i.e., atoms, molecules, solutions) react to light. Some studies depend on how much light an atom absorbs. The electromagnetic radiation absorbed, emitted or scattered by the molecule is analyzed. Typically, a beam of radiation from a source such as a laser is passed through a sample, and the radiation exiting the sample is measured. Some, like Raman, depend on a molecule's vibrations in reaction to the light. Sir Chandrashekhara Venkata Raman November 7, 1888-November 21, 1970 Won the Noble Prize in 1930 for Physics Discovered the "Raman Effect" Besides Discovering the Raman Effect, He studied extensively in X-Ray Diffractions, Acoustics, Optics, Dielectrics, Ultrasonics, Photo electricity, and colloidal particles. SERS vs. Raman During charge transfer, some vibrations will be altered due to a molecule's interaction with the surface, resulting in some of the SER bands being shifted in respect to the normal Raman Spectrum. SER spectra of a different small peptides and proteins were obtained from a roughened surface using a Raman spectrometer. Why Raman? In Raman spectroscopy, by varying the frequency of the radiation, a spectrum can be produced, showing the intensity of the exiting radiation for each frequency. This spectrum will show which frequencies of radiation have been absorbed by the molecule to raise it to higher vibrational energy states. What Exactly Is Being Measured? METHANE When Light hits a sample, It is Excited, and is forced to vibrate and move. It is these vibrations which we are measuring. Stokes vs. Anti Stokes Atoms are at a certain energy level at any given time. As a laser light hits the atom, it is excited and reaches a higher level of energy, and then is brought back down. If an atom is at a given energy level, it can be excited then fall below the original level. Anti-stokes spectrum are mirror spectrums of Stokes Raman Spectrums Using Stokes/Peaks to Determine Vibrations How To carry Out a Raman Experiment An insulated conductor was soldered to a piece of pure silver which was the embedded into a chemically resistant resin, leaving one face exposed. 1 micro liter of sample solution was placed on the roughened surface. The Sample is put through an Oxidation Reduction Cycle. Raman Instruments Results of Other Raman Studies Proposing a Structure Histidine One of the common natural amino acids. Nutritionally, histidine is considered an essential Amino Acid, but mostly only in children. In Catalytic Triads, the basic nitrogen of histidine is used to extract a proton from Serine, Threonine or Cysteine. Histidine is also used as a ‘Proton Shuttle’ Our Experiment To obtain an acceptable SERS Spectrum of Histidine Comparison of Spectrum to IR and Raman Spectrum Proposing a structure for Histidine Solid Histadine 13000 12500 12000 11500 11000 Wavelength 10500 10000 9500 9000 8500 8000 7500 7000 6500 6000 1 101 201 301 401 501 601 701 801 901 1001 1101 Wavelength 1201 1301 1401 1501 1601 1701 1801 1901 Histadine 3000 2800 2600 2400 2000 Wavenumber 2200 Series2 1800 1600 1400 1200 1 101 201 301 401 501 601 701 801 901 1001 Wavelength 1101 1201 1301 1401 1501 1601 1701 1000 1801 Citations And References Stewart, Fredricks, Surface Enhanced Raman Spectroscopy of Peptides and Proteins absorbed on a Silver Surface on an electrochemically Prepared Silver Surface, Spectrochima Acta, Part A, 55, 1999, pg. 1615-1640 Roth, Hope, Fredricks, et al. Simple Technique for measuring Surface Enhanced Fourier Transform Raman Spectra of Organic Compounds. Applied Spectroscopy, Vol. 47, 11, 1993. Habucchi, Cotlet, Gronheind, et al. Single Molecule Surface Enhanced Resonance Raman Spectroscopy of the Enhanced Green Fluorescent Protein. Journal of the American Chemical Society, 2003, Vol. 125, pg. 8446-8447 Acknowledgements Dr. Sat Bhattacharya HCS Dr. Ronald Birke Professor Tudor Fen Xu City College of New York