J Nucar Magnetic 10 Resonance ‘The phenomencn of nuclear magnetic resonance (NMR), first observed in 1946, deals with transitions between energy levels that arise because of the different orientations of magnetic moment of nuclei placed in a magnetic field. These transitions are studied by means of a resonance method. Hence, the name nuclear magnetic resonance. The technique was applied to problems in chemistry right from its discovery. With the advent of Fourier transform and other advanced techniques, it has become a scientific discipline of its own. 10.1 MAGNETIC PROPERTIES OF NUCLEI One of the most important properties pf a nucleus is its spin / or intrinsic spin angular morientum If, This gives rise to a magnetic moment to the nucleus. The following rules are useful in determining the value of nuclear spin J. (i) If mass number A is odd and atomic number Z is even or odd, / is half integer. Examples: 1H, '3C, ‘$n, '3 Gi) If A and Z are both even, J is zero. Examples: He, Cc, 0, 7S, (ii) If A is even and Z odd, J is integer. Examples: 7H, '?B, ''N, ‘The projection of the spin vector I on any direction can have the: values m= - 1 (1+ Demy E-U= DL dot) These are 2/ + 1 in number, and all are degenerate in the absence of an external magnetic field. In a magnetic field it splits up into 2/ + 1 states. The magnetic moment }1 associated with the spin angular momentum is given by pe ya (10.2) Here, 7 is a scalar called the gyromagnetic ratio and may take positive or negative value. An alternate expression for the magnetic moment is B= bytiy (10.2a) where gy, the nuclear g factor is a small dimensionless algebraic number, that takes different values for different nuclei, jy is the nuclear magneton and is defined by by ca = 5.051 x 1077 yT-!@Nm*) (10.3) where my, is the proton mass. 23Raswoana wee reba -hamo = 2b . Conve ahem, zr CN \ * [. ~ ‘ 820, - wait NMR experiment a vides, thamsitun Fem lower com ll ithe. oneagy em radia’ je? Bohr fey Condabion hy= Ha = QysHnbe > avmane cond ken | typical mag. falds oxi nine 228 Ey oT 22> fall under radio {uqueny reqen Source uneel © todince Qdmibon ov Affent From ohn Sagions of Spectroscopy - Resonamer cendihen —> acheined by applyeng escalating at doamoe’ sg; Lat = dormer Aesquoney > Ag at udnich magneké manent of “uel " Precns anound external maz Rul es erailotng M4 a patasad fA appl ig an orullabg vi te ae whee axu @ vet ‘4 ea. 1 Pe anlerra) Stabe {i proten >cornremy vast all nde ; Sete oa tae eeZoNANCE CONDITION 4/7 wooess of magnetic moment 41 is placed in a magnetic field By. the inert E = -p-By =~ =~ Bam By = ~ MBy cos 0 = - HB m wie Since m, can have 2/ + | velues, we will have 2/ + 1 equally spaced energy level ‘energy separation between any two adjacent levels is given by ~ in WOE] = By SPL = HP ew ay ny By ‘The basis of NMR experiment is to induce a transition from a lower level to the next higher level. If v is the frequency of the electromagnetic radiation that induces transitions between adjacent energy levels, Bohr’s frequency condition is then va BO wanna con Equation (10.5) is the resonance condition. For a spin 1/2 system we will have 2 states, ode corresponding t m, = 1/2 and the other for m, = -12. ‘Their energies are Ey, = “Hy and Eg = +B 06 ‘The resonance condition reduces to ‘These are represented in Figure 10.1. For proton, 11 = 2.79268 jy, and therefore, x 268 x (5051x1077 JT" 6 gly ya 22219268 (5051107 Tp, 5 ap sa x 108 Te) By (107) 6626x107 5 m7 12: Ey” Bo E218 \ m= U2; Ey =-1B eae 5 " in = -HBy Figure 10.1 Energy levels of a spin 1/2 system in an external magnetic field B = By.Nuclear Magnetic Retonance 218 netic fields used in NMR experiments are in the range 1 to 5 yr. a tee the nmeters for some of the nuclel are listed in Table 10.1 The erent ina ytorequency region of the electromagnetic spectrum and the SSS '¢ transitions are different from those used in the other tegions of oscillating, at the Ps onmnee condition is achieved by applying a radiofrequency field Oscillating ele sg armor frequency perpendicular to the extemal magnetic field, ‘This ng field is produced by applying an oscillating voltage to a coll whose axis is Perpendicular to the external static ficld. Proton is one of the most important nuclei having a spin J = 1/2. It is a constituent in almost all organic molecules. The NMR of a proton nucleus is often called Proton Magnetic Resonance (PMR). Another nucleus which is present in all organic compounds is carbon, As "C has no magnetic moment, it will not show any NMR spectrum, However, °C has 4 magnetic moment and shows resonance. Table 10.1 Magnetic Resonance Parameters for Certain Important Nuclei Nucleus Nuclear spin Magnetic moment v(MHz) (units of Hy) By = 1T 3T 5sT ‘ny Rn 2.7927 42.577 127.728 218.88 el 12 0.7022 10.705 32.115 53.525 OR wm 2.6273 40.055 120.165 200.275 oh) w 1.1305 17.236 51.708 86.180 *H 1 0.8574 6.536 19.608 32.680 MN 1 0.4036, 3.076 9.228 15.380 “B 3 2.6880 13.660 40.980 68.300 Se, 32 0.8209 4172 12.516 20.860 2G 3 0.6833 3.472 10.416 17.360 Br an. 2.0991 10.667 32.001 53.335 “Br 3a 2.2626 11.499 34.497 57.495 NMR was first discovered in (1946) independently by Purcell, Pound and Torrey in paraffin and by Bloch, Hansen and Packard in water. It is interesting to note here that the famous Dutch physicist C.J. Gorter (1942) was the first person to look for nuclear magnetic resonance bulk matter. Due to his bad luck he failed to observe resonance as the sample was easily saturated because of its large spin-lattice relaxation time (Section 10.5). 10.3 NMR INSTRUMENTATION We have already seen that the basis of NMR experiment is to induce a transition between two consecutive energy levels that result when a nucleus is placed in a magnetic field. The resonance condition is given by Eq. (10.5). One can observe the spectrum experimentally either by varying the external magnetic field keeping the frequency fixed or by varying the frequency, keeping the magnetic field fixed. The first approach is often ‘preferred since it is difficult to vary the frequency at a very high level of stability (one in 10? or better). hve Foctee — Aenama condubion© requirements of a typical NMR spectrometer are: (@ An electromagnet giving a powerful, stable and homogeneous magnetic field, The field: must be constant over the area of the sample and over the period of time of the experiment. (i) A sweep generator which supplies variable current to a secondary magnet, Then _.. the total applied magnetic field can be varied over a small range. (ili) The sample container, Usually a glass tube (5 mm OD) spun by an air-driven turbine to average the magnetic field over the sample container. The process is often referred to as the spinning of the sample, GW) A FF oscillator connected to a coil, called the transmitter coil, transmits the energy (o the sample. As mentioned, the axis of the coil has to be perpendicular to the magnetic field, (A ff receiver connected to a coil, called the receiver coil, encircles the sample. Its axis has to be perpendicular to both the magnetic field and the axis of the Wansmitter coil. The transmitter and receiver coils and the sample holder are Constructed into a single unit called probe. Often the necessary small magnetic Sweep field is obtained by use of sweep coils built into the NMR probe. (vi) A read out system consisting of an r-f amplifier, recorder and other accessories to increase the sensitivity, resolution and accuracy. A schematic representation of these components are shown in Figure 10.2. The magnetic field at which the resonance occurs at a fixed frequency for a given nucleus is constant. Therefore, each instrument normally works only for a particular kind of nucleus. Several nuclei can be studied by changing the oscillator; obviously, the field has also to be changed accordingly. In the case of solids, since the width of lines are expected to be large, this type of high resolution investigation is not possible, RE receiver} From receiver To transmitter i coil coil wate Figure 10.2 Block diagram of a NMR spectrometer.10.19 NMR IMAGING Nuclear Magnetic Resonance Imaging (NMR) is a new diagnostic technique which has revolutionised medical imaging and biomedical research. Unlike its predecessor, X-ray izing electromagnetic computerised tomography (CT scanning), it does not_u: but rather uses interaction between a magnetic field and magnetic nuclei. There are only very few restrictions for NMRI since it is considered risk-free for all patients except those- with extensive life-support systems.peck. press -18 WD angular vde Ww Nuclear Magnetic Resonance 303 NMR, NMRI is based on the fundamental relation @ = 7B, Eq. (10.14). Like ena ee in one tesla magnetic field along the z-direction, the Proton resonates at ae NMRI has focussed mainly on proton (hydrogen nucleus) because hydrogen is ms abundant element in the human body and is highly sensitive to Fesonance. The biological samples, tissues, etc, contain water Or olier Maids wih prions Pee cee magnetically-active nuclei "°F, Na, *'P, “N can also be imaged. In NMR, the resonance absorption is recorded for very small bulk sample by lying a homogeneous field over it, In NMRI, one has to get the resonance of normal tissues and the pathological state of the ‘issues over a region of interest: That is, one has to measure the net magnetization at each location in the object. This is done by ‘spatial -theoding’, encoding the signal in such a way that its spatial dependence can somehow be decoded. This is achieved by the use of magnetic gradients produced by suitably- Placed electrical coils. Gradients give rise to a spatially-dependent distribution of NMR frequencies. Gradients used for imaging are generally in the 1 gausS/em Tange. thereby Producing magnetic fields considerably smaller than that of the primary field. NMRI is thus accomplished by making the field inhomogeneous. The conventional high resolution NMR uses homogeneous Magnetic fields. When the magnetic fiel i ically across an object, the resonance frequency Of a normal nucleus and the one affected disease appear at diferent magasie fas as Pips 1st For apie elcas increased iron deposition in the brain such as the globus pallidus, substantia nigra, red nucleus and denate nucleus show a different signal from that of the remainder of the brain. net 2 Part of brain Affected region oe 2 5 Frequency ali, Figure 10.16 The basic principle of NMR imaging. B, is the field required for signal from the affected region and B, for the signal from the normal region. vMolecular Structure and. ‘Spectroscopy The contrast parameters normally used for NMRI are spin density, rela: ation times T, and T;, chemical shift and the fluid flow velocity of the object. We shall consider the Parameter, relaxation time 7, in detail. Relaxation time vary over a wide range of values (0.3 to 1.28 at 100 MHz) and are dependent on the pathological state of the tissues. Therefore, relaxation times are often preferred as contrast agents for the discrimination of anatomical features. As already seen, spin-lattice relaxation time 7, characterizes the time-dependent return of the z component of magnetization M, to Mo, the equilibrium value. If a spin system is subjected to a 90° pulse, Man = mi -exp (#)) (10.73) The spin-spin Telaxation time T, governs the decay of any transverse magnetization to zero. Again for a 90° pulse M,,(t) = Mo on) (10.74) The different imaging methods are classified according to the spatial element that is detected in each elementary step of the sequence. The common ones are, (i) sequential point, (ji) sequential line, (iii) sequential plane, and (iv) simultaneous volume method. Sequential point imaging involves obtaining the NMR response from only a single point in the sample at any one time. The image is then formed by sequential scanning of the selected point over the entire region of interest. In the sequential line method, a line-by-line scan is Performed through the region of interest. This method is more efficient and sophisticated than the previous one. In a similar way an image can be produced by selecting sequential planes or by simultaneously imaging the whole volume. Consideration regarding the SIN _ ratio, freedom from image distortion, minimum imaging time, etc. decides the selection of ~ the technique. - The field of NMRI continues to be very fertile for both technical research and clinical development of new applications. Although it is used mainly for medical applications, its potential for nonmedical applications has now been recognized.Electron Spin Resonance 1 1 14.1 INTRODUCTION Electron spin resonance (ESR), anperetrscopie technique confined to the snes i ‘The ‘magnetic moment arises from spin Paramagnetic systems, the most sition metal ions, fee radicals, ions and molecules having an odd sPin and charge of the electron confer on it a spin magnetic moment j1, given by Eq. (3.13) rn ‘THE orbital magnetic moment jy is given by 312Electron Spin Resonance 343 the a els babies i sad 1. As Won 9 =) ,t, sobmunlitg tw vale or from E = guiyBm, any Re may be noted equal to th fer ey that we considered only the spin magnetic moment. Since spin ¥ is ton, m, = £14 and we get two levels with energies (Figure I1-1) 1 Bas -Sege, Bye Senne V2 Bg fame AE gin NS ato) ay =I Byg*=fanyB Figure 1.1 Zeeman splitting of an unpaired electron in a magnetic field B. IF electromagnetic radiation of frequency v satisfying the condition AV = Ey ~ Egy = SHB is present, transition between these Zeeman levels occur which is studied by ESR. Equation (11.8) gives the resonance condition for ESR observation, For free electron, g = 2.0023. In a field of 0.34 T, from Eq. (11.8) v = 2:0023(9.274 x10 IT"034T 9595 winte 6.626 x10" J. ‘This frequency falls in the microwave region. Hence, microwave source and techniques have to be applied for the observation of ESR. ‘Successful observation of the ESR spectrum requires suitable values for the frequency V and the magnetic field B to satisty Eq. (11.8). For magnetic field of 0.34 T, v comes out to be about 9.5 GHz. Spectrometers operating around this frequency are often referred to as X-band spectrometers. Spectrometers operating at higher magnetic fields (B = 13 T) with corresponding higher microwave frequency (~ 35 GHz) are also available. They are called Q-band spectrometers. 11.3 ESR SPECTROMETER Basic requirements: The basic relation for the observation of ESR transition is the one given by Eq. (11.8). For continuous absorption one can either_v uency_V across res0- ance keeping the field B constant or the magnetic field is varied keeping the frequency a8)Tater keeping the stability oe ne '8 usually preferred since it is easier to vary the magnetic field very high levels. In electromagnetic radiations, the electric Meld, the Id and the direction of Propagation are mutually perpendicular. In ESR experi- im . 1 Magnetic Component of the electromagnetic radiation that interacts with the etc dipole ested wi npaired elecion. If the oscillating magnetic field vector even (aélttion is parallel tthe applied magretic Neld, only un celine ot we energy ea levels will pe" 2°! & Feorientation of the electron magnetic moment. Transition between levels will fg take place only when there is a reorientation of the electron magnetic moment. fore, f Or successful observation of ESR absorption, the oscillating magnetic field must ycillating magnetic 3 4pplied perpendicular to the applied magnetic field. Population in "be the lower level will be considerably high even for larger extemal magnetic fields. Though higher population in the lower level leads to larger absorption cross-section and higher sensitivity, one prefers to work with reasonable fields only as this corresponds eet hiantls Svity of fait large dimensions (2 x I x 4 em). Alo, itis more difficult to et highly homogeneous Magnetic fields of very high value over a larger region. Some of the basic requirements of a X-band ESR spectrometer are (0 An electromagnet capable of supplying a homogeneous magnetic field which can be ji) Sind linearly on either side of the magnetic field (> Source of microwave radiation in the region of 9.5 GHz (ii) Suitable sample cavity Gv) Arrangements for transmitting the radiation energy into the sample cavity 2 Detection system to measure the variation in microwave power (Wi) Suitable oscilloscope or recorder. Description of the set-up: simple block diagram of a balanced bridge ESR spectrometer is given in Figure 11.2. The usual source of radiation is a Klystron oscillator which produces Monochromatic radiation of the required frequency. The radiation from the source is “ransmitted to the sample cavity through a microwave impedance bridge. The rectangular Dummy load Crystal Klystron Phase sensi- tive detector| ‘Modulator L Recorder] Figure 11.2 Block diagram of a simple balanced bridge ESR spectrometer. To modulating coils. Electron Spin Resonance 315 Rlectromar, SAvity which contains the sample is kept in between the pole pieces of the fourth anes A Sumimy load is kept In the third arm and a semiconducting crystal in the Am Of the microwave bridge. The radiations that arrive in the dth arm are detected by the crystal. It is then amplified and fed to a suitable recorder, Phase sensitive detector ate usually used to detect ESR signals and represented as absorption or first derivative Curves (Figure 11-3). The magnetic field is Swept over a small range across the resonance Condition by varying the current in a pair of sweep coils mounted on the cavity walls. (a) ©) / f Mv . ) @) Figure 11.3 ESR signal: (a) a ‘single absorption line, (b) its first derivative, (c) four equally- spaced overlapping absorption lines, (d) first derivative of the spectrum in (c). ‘Working: When the bridge is in a balanced position, microwave power flows only in the two arms—the one to cavity and the other to the dummy load. There will not be any power in the fourth arm. Power in the fourth arm will be there only when the bridge is not balanced. Thus, if balance exists, initially no signal appears at the detector an¢ when the sample absorbs, the balancing of the bridge is lost and power appears in the fourth arm. The width of ESR lines are fairly large and hence the spectrum is usually recorded in the first derivative mode which enables one to fix up the frequency position and estimation of intensity more precisely. Another advantage of derivative mode is that it gives a well-defined linewidth AB (Figure 11.3). Even if there are overlapping signals, it is still possible to do a good estimate of AB.