Chemistry 125: Lecture 59 March 21, 2011 Precession and MRI NMR Spectroscopy Chemical Shift This For copyright notice see final page of this file 90° RF Pulse and the “Rotating Frame” Fast precession (~100 MHz) Slow precession (~0.1 MHz) Applied Magnetic Field Until “relaxation” 100 MHz RF reestablishes in lab frame equilibrium. Precessing proton gives rise to constant vertical field and rotating horizontal field. Will rotating horizontal field generate 100 MHz RF signal? No, because there are many precessing protons with all possible phases. Horizontal fields cancel. Consider a “rotating frame” in which the observer orbits at 100 MHz - protons seem to stand still as if no applied field. Pulse a very weak magnetic field fixed in this rotating frame (just long enough to rotate all nuclear spin axes by 90°). Subsequent precession generates 100 MHz RF signal in lab frame. A 90° pulse makes spinning nuclei (1H, 13C) “broadcast” a frequency that reports their local magnetic field. MRI: locating protons within body using non-uniform field X-Ray Tomography www.colorado.edu/physics/2000/tomography/final_rib_cage.html MRI: find protons in body (e.g. fluid H2O) So there are protons in the body, but where? ~1.5 Tesla (15,000 Gauss) Bz wrap in several miles of special wire at 4K protons precess at 63 MHz Superconducting Solenoid How to locate Crickets, if you can’t see them: Establish a temperature gradient and listen with a stopwatch. P. LeMone (2007) MRI: find protons in body (e.g. fluid H2O) These three gradients subtract So there are protons inallow the body, Four analogous dB /dy slicing in all directions z from B but where? subtract top/bottom zcoils to construct aestablish 3D on left from Bz const Hz tomograph. near feet ~1.5 Tesla (15,000 Gauss) Bz add to Bz add to Bz near head on right protons precess at 63 MHz Superconducting Solenoid Functional MRI: locating protons whose signal strength is being fiddled with BOLD Imaging minus Subject recently fed Subject Fasting Functional MRI (fMRI) e.g. Blood Oxygen-Level Dependent (BOLD) Imaging Spatial Resolution ~1 mm Temporal Resolution 2 sec with permission of Dr. Tony Goldstone, Imperial College Difference Map Cell activity increases blood oxygen supply, speeds relaxation. NMR: locating protons within molecules using uniform field ? Fractional Listen at fixed frequency. difference in Tune Bo to “hear” precession. applied field 2.48 ppm 0.00000248 ! Requires very high uniformity of field to avoid “MRI” The “Chemical” Shift HO-CH2-CH3 http://www3.wooster.edu/chemistry/is/brubaker/nmr/nmr_spectrum.html Bo Oscilliscope Trace (1951) In the late 1950s chemistry departments began buying NMR spectrometers with fields homogeneous enough to determine molecular structures from chemical shifts (and spin-spin splittings). With multi-user equipment, it was a challenge to keep the fields sufficiently homogeneous to obtain sharp lines. At SUNY-Stony Brook in 1972 physical chemist DoLauterbur Not Touch Gradient Knobs!!! Paul wouldThese take over the departmental or inhomogeneity. this one! machine nightly and destroy the field By establishing gradients in different directions he located two 1 mm tubes of H2O within a 5 mm tube of D2O, and published this “zeugmatogram” in Nature in 1973. 30 years later he shared the Nobel Prize in Physiology or Medicine for inventing MRI. Magnetic Resonance Spectrometers (and X-ray Diffractometers) have put classical structure proof by chemical transformation (and even IR!) out of business. One Yale “natural products” organic professor, whose research used chemical transformations to puzzle out molecular structures, abandoned organic chemistry to take up fundamental research on quantum theory (and later became a professional studio photographer). Some of the Magnetic Resonance Spectrometers in Yale's Chemistry Department 500 MHz 500 MHz 600 MHz 600 MHz 800 MHz * 3 ~8 = 512 times as sensitive as 100 MHz (not to mention the chemical shift advantage discussed below) * 1) Boltzmann factor 2) Energy quantum 3) Electronics sensitivity EPR (Electron Paramagnetic Resonance) (for Free Radicals with SOMOs) e magnet is 660x H+! EPR (Electron Paramagnetic Resonance) 9 GHz ~3000 Gauss (0.3 Tesla) New 1000 MHz (23.5 Tesla) NMR Spectrometer NHFML - Florida State University now has a pulsed field NMR at 45 Tesla (there is no charge for use, but you have to have a great experiment Which peak is which set of protons? 1 2 Area (integral) 3 HO-CH2-CH3 http://www.wooster.edu/chemistry/is/brubaker/nmr number of protons, because they are so similar (not like IR) Oscilliscope Trace (1951) O O O ? O O O O O 3:1 1:1 1) O3 2) H2O2 ? Structural proof by chemical HO-C C-OH degradation 1955 Advertisement O O cis-caronic acid (venerable) C H 2.9 1 H C http://www.wooster.edu/chemistry/is/brubaker/nmr O CH3C Peak Width ~3 ppb OCH2CH3 Triplet (1:2:1) in CDCl CHCl3 solvent at 5.9T (250 MHz) 0.029 ppm High Resolution “Low” Resolution × 250 MHz (~3 ppb, (~0.3sample ppm) spun) Quartet 7.3 (1:3:3:1) 7.3 8 7.3 7.3 3 2 ? 7 6 5 4 3 d (ppm) Chem 220 NMR problem 7 7.3 Hz 2 3 1 0 A 90° pulse makes spinning nuclei (1H, 13C) “broadcast” a frequency that tells their LOCAL magnetic field. Components of Effective Magnetic Field. Bmolecular (diamagnetic) Applied Field: Inhomogeneous ~ 30,000 G for MRI CAT scan. (4 G/cm for humans, 50 G/cm for small animals) Bapplied Homogeneous for Chemical NMR Spectroscopy Beffective (spin sample) Molecular Field: Net electron orbiting - “Chemical Shift” (Range ~12 ppm for 1H, ~ 200 ppm for 13C) Nearby magnetic nuclei - “Spin-Spin Splitting” (In solution JHH 0-30 Hz ; JCH 0-250 Hz) The Chemical Shift high electron density Chemical Shift and Shielding Note: Electron orbiting to give B is driven by B; so B B. CH3 d+ dSi H3C CH3 H3C Bmolecular (diamagnetic) R-OH (depends on conc, T) O RC O RC OH Bapplied Beffective 11 H H 10 9 deshielded downfield low e- density high chemical shift high frequency 8 7 CH3C C C H 6 TMS X = O, Hal, N O RC CH Alkyl X CH R-H 5 d (ppm) C-H ? 4 3 2 1 ! ??? shielded upfield high e- density low chemical shift low frequency 0 Bapplied Diamagnetism from Orbiting Electrons End of Lecture 59 March 21, 2011 Copyright © J. M. McBride 2011. Some rights reserved. 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