S14, NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY INTRODUCTION The nuclei of certain elements and isotopes behave as though they are magnets spinning about their axes. The nuclei of ordinary hydrogen ('H), carbon-13 (°C), fluorine (°F), phosphorus (*'P), etc. possess this property. When a compound containing 'H or °C atoms is placed in a very strong magnetic field and simultaneously irradiated with electromagnetic energy, the nuclei of the compound may absorb energy through a process called magnetic resonance. This absorption of energy is quantized and produces a characteristic spectrum for the compound. Absorption of energy does not occur unless the strength of the magnetic field and the frequency of electromagnetic radiation are at specific values. The NMR spectroscopy deals with the transitions that are induced between the magnetic energy levels of the nuclei of the molecule by radio frequency (RF) waves. The magnetic energy levels are created by ‘placing the nuclei in the external magnetic field.Be thod ofNMR wee eeleneeeEeE=S This method of NMR was first developed by Edward Purcell and Felix Block in 1946 and were awarded Nobel prize for their valuable discovery. Since then, NMR spectroscoPY has developed into a most powerful tool for structure elucidation and identification of organic molecules. MAGNETIC PROPERTIES OF NUCLEI Like electrons, the particles of the nucleus, i.e., protons and neutrons are also considered to spin about their own axes and each particle of the nucleus has a spin quantum number of A. If the protons and neutrons ina nucleus are not paired, the nucleus as a whole will have a resultant nuclear spin, I. The numerical value of this nuclear spin quantum number, I, depends upon the number of protons and neutrons having: parallel and anti parallel Spins. It is related to the mass number and atomic number as follows: ‘Atomic mass Atomic number Nuclear spin quantum number, I i 13 Odd Even or Odd hat 272? Even Even 0 Even Odd In agreement with the above general rules: () Nuclei such as |H (proton), 7H, {C, 7N, oF, jsP, ete. all have I= 3; 8 ” "B, Na, 7 CL ® Br, 8 Br, etc. haveI= 3 and O has been found to have I= 5. (i) Nuclei suchas $He, 7 C, O, 4g Sallhavel-0. Itis fortunate for organic chemists that ® Cand '* O are non-magnetic, otherwise the NMR spectra of organic molecules would have been much more complex. (ii) Nuclei such as ; H (deuterium) and \ N have I=1 while '? B has I=3.Since an atomic nucleus is associated with charge, therefore, ee any Totating charge, it generates a magnetic field along the axis ofits spin. Therefore, suc a nucleus behaves as a tiny bar magnet and has a characteristic magnetic moment, uw. | | | NUCLEAR SPIN STATES | When a nucleus with I> 0 is placed in an external magnetic field, it is found to have a set of discrete orientations w.r. the applied magnetic field. These various orientations are referred to as spin states. For a nucleus with spin quantum number ]> 0, there are 21+1 allowed spin states which differ from one another by unity, i.¢., the various spin states are —1,(-1+1) (1-1), +1. Thus, it follows that nuclear spin states like electronic and vibrational states Gre quantized. For example, a proton (hydrogen nucleus) with I= +, has two allowed spin states, 2(1) +1 = 2 designated as — 1 and +4. For chlorine with I= 4, there are four allowed spin states, 2() +1 = 4 designated as — 3, — 4,ttand+3. In the absence of a strong external magnetic field, the nuclear spins of magnetic nuclei are orientated randomly (Fig 5.15 a). However, when these magnetic nuclei are placed between the poles of a strong magnet, they either align with the field or ‘oppose the field (Fig 5.15 5). Fig 5.15. Orientation of nuclear spin : (a) Random orien external magnetic field, H, and (6) Specific orientation, i magnetic field, H,. tation, in the absence of an in the presence of an external When a compound containing hydrogen (and therefore applied external magnetic field, the protons may assume o: orientations w.r.t the external magnetic field (Fig 5.16). These t protons) is placed in an ne of the two possible wo possible orientations(spin states) do not have the same cnergy and hence are not equally populated. The orientation in which the spins of the nuclear magnets are aligned parallel (I= +4) to the extemal magnetic field is slightly lowerin energy and hence this spin state is slightly favoured over the other orientation (I= — 4) in which the spins of the nuclear magnets are aligned anti parallel to the external magnetic field (Pig 5.16). » | ¢ Spin + 1/2 aligned with in —1/2 aligned opposite the applied field Spin 9 Direction of the to the applied field applied magnetic field, H, Fig. 5.16. The two allowed spin states ofa proton. The difference in energy between the two spin states is designated as AE. In general, the value of AE is very small but the magnitude depends upon the strength of the applied magnetic field as shown in Fig 5.17. _(1=-1/2) 3 i AE,AE,>AE, (le+172) Strength of the applied, H—> H,=23,500 G Fig5.17. Variation of energy separation between nuclear spin states as a function of applied field strength. It is quite clear from the Fig. 5.17 that when H,=0, AE=0, i.e., in the absence of amagnetic field, the two spin states of a proton have equal energy. However, as the strength of the applied field increases, the energy difference between the two states increases. Ata field strength of 14,100 G or 1.41 T, AE = 2.39 x 1075 kJmol™! while when H = 23,500 G or 2.35, AE = 3.97 x 107 kJmol~! (where G = Gauss and T= Tesla are units of magnetic field in CGS and SI units respectively and further 1T=10°G).5.15. PRINCIPLE OF NUCLEAR MAGNETIC RESONANCE _ _ Tounderstand the principle of nuclear magnetic resonance, let us consider the motion ofa common spinning top. When the spinning top is disturbed a little, its spinning axis will trace out a circle perpendicular to the earth’s gravitational field as shown in the Fig 5.18(a), Such a motion is called precession, }«— Precessional orbit Axis of rotation Spinning nucleus t t t 4 t t fr direction of magnetic field x Fig 5.18.(a) Spinning top precessing in the earth's gravitational field : (b) the precession ofa spinning nucleus due to the applied magnetic field. Ina similar way, when a nuclear magnet is placed in an external magnetic field, it will experience torque (force perpendicular to the axis of the nuclear magnet) which will tend to align it with the field. Since the nuclear magnet is spinning, it will not align with the external magnetic field but will precess around it (Fig 5.18 ) with a certain angular frequency @, called the precessional | frequency or the Larmor frequency. The frequency at which a proton will precess, depends upon the strength of the applied magnetic field. Stronger the applied field, higher is the precessional frequency. For a proton, if the applied magnetic field is 14100 Gauss, the precessional frequency will be 60 MHz (60 million cycles per second). Since a proton has two allowed spin states (I=+1 and 1=— +), it can precess in two principal orientations—one aligned with the applied field (low energy) and the other opposed to the applied field (high energy) as shown in Fig 5.16. It is possible to induce transitions between these two orientations by irradiating the precessing nuclei (protons) with radiowaves (electromagnetic radiations) of correct frequency. When the precessional frequency of the nucleus (proton) is exactly equal to the frequency of the electromagnetic radiations, the absorption occurs and the nuclei in lower energy spin state ‘flip’ to the higher energy spin state. When this occurs, the nucleus and the radiofrequency (RF) beam are said to be in resonanceTa and since this resonance is due to magneti i aaa i eri magnetic resonance (NMR), ‘ic properties of the nuclei, it is called nuclear as Sei amount ofthe radiofrequency energy needed for resonance depends ae external magnetic field and on the nucleus being irradiated, i.e., 5 A AE =hv=y aH Where AE = Energy difference between the two spin states, Ah =Planck’s constant H = Strength of applied field in gauss. ¥ = Gyro magnetic ratio. {itis ratio of magnetic moment to angular momentum and is constant for each nucleus. For a proton, the value of = 26,750) It may, however, be noted that a NMR instrument with a magnetic field strength of 1.41T and operating at a constant RF of 60 MHz, can induce transitions among proton spin states in a molecule and is not useful for other nuclei. Therefore, separate instruments are required to observe transitions in the nuclei of other elements such as carbon, fluorine, phosphorus, etc. However, now-a-days, the modern more expensive fourier transform (FT) instruments can be used to observe the resonance of nuclei of several different elements in a single instrument. Instruments with 200 and 300 MHz are now quite common and instruments with frequencies upto 800 MHz are found in the advanced research institutions and bigger universities. 5.16. PMR SPECTRUM-ORIGIN OF SIGNAL In principle, a PMR spectrum can be obtained by placing the substance containing hydrogen nuclei or protons in a magnetic field of constant strength and passing electromagnetic radiations of varying frequency through the substance and observe the frequency at which radiation is absorbed. In practice, however, it has been found more convenient to keep the frequency of the electromagnetic radiation constant and vary the strength of the magnetic field. At some value of the field strength, the energy required to spin the proton matches the energy of the electromagnetic tadiations. At this stage, absorption occurs and a signal is observed. Such a spectrum is called PMR spectrum. ‘The PMR spectrum is a plot of intensity of absorption along y-axis against magnetic field strength along x-axis. By convention, magnetic field strength increases from left to right across the chart. Since the magnetic field strength and the frequency are related by the equation,hl 2m 'y absorption signal along the x-axis can be expressed in either field or nits. A typical NMR absorption signal is shown in Fig 5.19. the position of an the frequency wr +—Downfield direction Signal Intensity of absorption —> Cow Fred Tigh Field Magnetic Field Strength —> Fig. 5.19. Atypical PMR signal We have discussed above that a PMR signal is obtained at some value of the field strength at which the energy required to flip the proton becomes equal to the energy of the electromagnetic radiation, Therefore, we may expect that all the protons in an organic molecule will absorb at exactly the same field strength. Consequently, the PMR spectrum would consist ofa: single signal and it will be of little use to the organic chemist. In practice, ithas been found that all the Protons do not absorb at the same frequency. This is due to the reason that the frequency at which a proton absorbs depends upon the magnetic field which that proton actually feels and this effective field strength is not exactly the same as the applied magnetic field. This effective field strength which a proton feels depends upon its local environment, i.e, electron density around that proton and the Presence of other protons in its neighbourhood. In other words, each proton or more precisely, each set of equivalent protons will have slightly different envi ixonment than every other set of equivalent protons and hence will absorb at slightly different applied field strength to experience the same effective field strength at which the absorption occurs. Thus, it follows that at a given radiofrequency, the protons having different environments experience the same effective field strength but absorb at different applied field strengths. Since in a PMR spectrum, absorption is plotted against the applied field strength, a PMR spectrum will consist of a nu ber of absorption signals which would be very useful in determining the structure of molecules, 5.17. APPLICATIONS OF PMR (NMR) SPECTROSCOPY PMR spectroscopy is widely used in the stud: ly of organic chemistry. Some of its important applications are discussed below :or 8anic compounds, Like IR spectrum, PMR spectrum . Print of the compound. If the PMR spectrum of a given sample a rad fi same as that ofa known compound, the given compound must be identical wil nown compound. In this way, PMR spectroscopy is used to identify the organic compounds, 7 {Identification of theo can also be used as a finge! 2, _ Determination of molecular structure or structural diagnosis. PMR spectra are often very complex. But this complexity isnot a disadvantage but is often useful in the determination of molecular structure as it makes the spectra characteristic. Furthermore, even the complex spectra can be interpreted by using the techniques of deuterium Iabelling and double resonance. Thus, PMR spectroscopy is increasingly being used asa versatile tool for structure determination of organic compounds. 3. Qualitative analysis, With the help of PMR spectrum of a compound, it is possible to detect the presence or absence of functional groups due to their characteristic chemical shifts. For example, appearance of a signal at 6 9.5 is indicative of -CHO group. 4. Identification of reaction products, Whenever in a reaction, two or more products are theoretically possible, PMR spectroscopy has been used to identify the actual product. For example, addition of HCI to 1-methylcyclohexene can, in principle, yield either product (I or II). CH, cH, CH, or +HCl —> ie cl or ee T W cl 1-Methyleyclohexene 1-Chloro-I-methyleyclohexane _1-Chloro-2-methyleyclohexane PMR spectrum of the product shows a singlet at about 61.5 for the methyl protons. This confirms that the product formed is 1-chloro-1-methylcyclohexane (I). Had the product been (II), it would have exhibited a doublet (J = ~7-8 Hz) for the three methyl protons at around 61.0. 5. Detection of aromaticity. Protons attached to the system following (4n +2) electrons rule (i.¢.., Huckel’* rule) where n = 1, 2, 3... (whole number) are extremely deshielded due to the circulating sextet (ring current) of z electrons. As a result of this, the signals, for the aromatic protons appear at a very low field, i.e., high d values. Thus, from this, the aromatic character of ‘various compounds can be predicted.molecular H-bonding can be distinguished from intermolecular H-bonding as the former Temains unchanged on dilution and consequently, the PMR spectra usually remain unaltered Y Varying the concentration or temperature, On the other hand, at high concentration, 5 value changes (becomes high) as More and more hydroxyl protons tend to show intermolecular H-bonding. 7 Detection of electronegative atom or group. As it is already known that the clectronegative atom or group decreases the electron density in the vicinity of proton nucleus ahd thus causes deshiclding and the signal is shifted downficld.Thus, the 6 values (ort values) provide information about the presence of an electronegative atom or group. 8. _ Distinction between geometrical isomers. PMR spectroscopy can be used to distinguish between cis and trans isomers. The distinction is based upon the magnitude of coupling constant, £e., 6—14 Hz for cis-protons and 11-18 Hz for irans-protons. H H H. Nonct Non’ H cis, J=6-14Hz trans, J = 11-18 Hz 9. __ Importance in quantitative analysis. NMR spectroscopy is gaining importance for the quantitative analysis of the compounds. Equilibrium mixtures can be analysed when the proton signals of the components are well separated. In the NMR spectrum of pure ethanol (CH,CH,OH), a triplet is formed for the OH proton but when water is added in alcohol, then due to proton exchange, the triplet collapses to a singlet. The _ Position of this singlet depends upon the water content in alcohol. From the values of the chemical shift, the ratio of water and alcohol can be estimated by comparing with the known results. Keto-enol tautomerism has also been studied in the same manner, Consider acetyl acetone in equilibrium with the enolic form ° ° ° HO Il I Il | CH,—C—CH,—C—CH, CH,— C—CH = C—cu, (Keto) (Enol) The NMR spectrum of pure acetyl! acetone shows signals for the following kinds of equivalent protons :ll a=7.86T , ; “CHy—C~'CH,—C_ecH, b=6.35r (ketonic) Ketonic on? ° a= 8.037 b=-43 ¢ 4.4 1 (enolic) d= 7.877 | i “CH;—C—"CH—C_ioy, Enolic A distinet two proton singlet at 6.35 7 appears due to the ketonic form whereas all the eight protons in the enolic form are observed as four signals.Quantitatively, the number of enol protons relative to the keto protons yields the ratio as under. Enol protons : Keto Protons =8 :2or4:]. Thus, we say that 80% of enolic form and 20% of the ketonic form are present in the equilibrium mixture of ace! tylacetone, 10. | Magnetic resonance imaging (MRI). An important application of PMR spectroscopy in medicine today is a technique called magnetic resonance imaging or MRI. Just as NMR is viewed as a sort of image ofa molecule, MRI is the image of the section of the human body. Thus, MRI can be utilized to discriminate between healthy and diseased tissues of body. It is based on the fact that the protons pr i? efc. resonate at a given frequency. Since human bo each water (HO) molecule has two hydrogen nuclei, so images of the different parts of the body can easily be taken, In a diseased condition of body part, the distribution of water, fats, lipids, etc. alters and hence by using MRI, one can detect the diseased part of the body.