1 Given that: for 1H = 267.512 106 rad T-1 s-1 for 13C = 67.

67.264 106 rad T-1 s-1 for 19F = 251.667 106 rad T-1 s-1

1(i). What is a Larmor frequency? The frequency at which a particular nuclei resonates and precesses. = B0 B0 [ Larmor frequency, , is directly proportional to B0 ] Different nuclei will precess at a different frequency:
1

H 0 ppm to 50 ppm C 0 ppm to 200 ppm

13

Quantum mechanics requires that the magnetic moments are actually not statically aligned exactly parallel or antiparallel to the external magnetic field, B0. Instead, they are forced to remain at a certain angle to B0, and this causes them to wobble around the axis of the field at a fixed frequency. This periodic wobbling motion that the top assumes in a gravitational field is called precession. Thus, the magnetic moment vector of a nucleus in a magnetic field also precesses with a characteristic angular frequency called the Larmor frequency (), which is a function solely of (the magnetogyric ratio) and B0: = B0 The angular Larmor frequency, in units of radians per second (rad s -1), can also be transformed into linear frequency, (in reciprocal seconds, s -1; or hertz, Hz) by division of 2: precession = /2 = B0/2

The rate of this precession is proportional to the external magnetic field strength (B0) and to the strength of the nuclear magnet (). This resonant frequency is in the radio frequency range for strong magnetic fields and can be measured by applying a radio frequency signal to the sample and varying the frequency until absorbance of energy is detected.

Figure 1 1(ii). At 18.78 Tesla, what is the Larmor frequency for a 13C and 19F? Since Larmor Frequency, = B0 and for 13C = 67.264 106 rad T-1 s-1, For 13C: = B0 = [ 67.264 106 rad T-1 s-1 ] 18.78 T = 1.263 109 rad s-1 Since Larmor Frequency, = B0 and for 19F = 251.667 106 rad T-1 s-1, For 19F: = B0 = [ 251.667 106 rad T-1 s-1 ] 18.78 T = 4.726 109 rad s-1

1(iii). What is the energy difference between the two spin states of 14.09 Tesla? Energy difference between the two spin states of 19F: E = E (m = ) E (m = ) E = [ ( ) () ] [ h B0 ] / 2 E = [ h B0 ] / 2 Since: B0 = 14.09 T h = Plancks constant = 6.626 10-34 J s for 19F = 251.667 106 rad T-1 s-1

19

F in a magnetic field of

3 E = [ h B0 ] / 2 E = [ (251.667 106 rad T-1 s-1) (6.626 10-34 J s) (14.09 T) ] 2(3.1416 rad) E = 3.739 10-25 J

1(iv). Calculate the maximum NOE, , for proton-decoupled 19F NMR spectrum. The NOE is an example of polarization transfer because polarization of one set of nuclear spin states (1H nuclear spin states) results in the polarization of another set ( 19F nuclear spin states). The maximum Overhauser signal enhancement () is given by: = (irr / obs) irr and obs are the magnetogyric ratios of the irradiated nuclei ( 1H) and observed nuclei (19F) respectively. Since: irr = for 1H = 267.512 106 rad T-1 s-1 obs = for 19F = 251.667 106 rad T-1 s-1 Thus, the maximum NOE, , for proton-decoupled 19F [19F-{1H}] NMR spectrum: = (irr / obs) = [ ( 267.512 106 rad T-1 s-1 ) ( 251.667 106 rad T-1 s-1 ) ] = 0.5315 2. What are spinning sidebands? What are sidebands from C satellite peaks? Spinning sidebands: Spinning sidebands are peaks seen around 1H peaks which are related to the rate of spin of an NMR tube. Spinning sidebands are an unavoidable result of spinning the sample to improve field homogeneity. If the field has been well shimmed, such spinning sidebands rarely exceed 1 % of the height of the main peak. Moreover, they will always be separated from the main peak by exactly the spin rate in hertz. This is, in fact, the most direct way to measure the sample spin rate. Although the spinning sidebands can be completely eliminated by turning off the spinner, however, this would cause substantial broadening of the signal (with an accompanying decrease in signal height) because of the small inhomogeneities present in the magnetic field.
13 13

C satellite peaks? How to distinguish spinning

Figure 2: An NMR signal peak showing sidebands (*) and signal (S) to noise (N) ratio.
13

C satellite peaks: Carbon satellite peaks are small peaks that can be seen shouldering the main peaks in

an NMR spectrum and are most often encountered in proton NMR. In the example of proton NMR, these peaks are not the result of protonproton coupling, but result from the coupling of
1

H atoms to an adjoining

13

C atom. These small peaks are known as carbon satellite peaks as

13

they are small and appear around the main 1H peak, that is, satellite around them. Carbon satellite peaks are small because C only makes up about 1% of the atomic carbon content of carbon, the rest of the carbon atoms are predominantly NMR inactive Carbon12 (12C). Carbon satellite peaks always appear as an evenly spaced pair around the main 1H peak. This is because they are the result of 1% of the 1H atoms coupling to an adjoined
13

C atom to give a

wide doublet (13C has a spin of a half). If the main 1Hpeak has protonproton coupling, then each satellite will be a miniature version of the main peak and will also show this 1Hcoupling. For example, if the main 1Hpeak is a doublet, then the carbon satellite peaks will appear as miniature doublets, that is, one doublet on either side of the main 1Hpeak.

Figure 3: The 13C satellite peaks flanking a greatly amplified TMS singlet.

5 To distinguish spinning sidebands from 13C satellite peaks: The difference between spinning sidebands with the
13

C satellite peaks is that changing

the sample spinning rate does not affect their position. In fact, the satellites are due to one bond 13CH coupling. The 13C satellite peaks are independent of sample spin rate or instrument operating frequency. Whereas, spinning sidebands are related to the rate of spin of an NMR tube. 3. Why a 13C NMR of CDCl3 shows a triplet of equal intensity at about 77 ppm? M = 2I + 1 D represents 2H (deuterium) I = -1, 0, 1 .. M = 2I + 1 = (2 1) + 1 = 3 (Triplet) CDCl3 is a very common NMR solvent which gives rise to a triplet of equal intensity at about 77 ppm in its 13C spectrum. This is because, D represents 2H (deuterium), an isotope of hydrogen with I = 1, which can therefore adopt three [2nI + 1 = (2 1 1) + 1 = 3] spin states (M = m = 1, 0, 1) with essentially equal populations in a magnetic field. Thus, the carbon signal of CDCl3 will be split into three equally intense lines, a 1 : 1 : 1 triplet, with line spacing
1

JCD.

4. Predict the multiplicity and indicate the relative intensity of each 1H signal for:

Br -- Non-active nuclei for NMR C* = Chiral Center

Hb H Hc He Hf * BrCC COCCBr Ha Br Hd He Hf

Ha coupled to Hb Doublet Coupled to H Doublet of Doublets

Hb coupled to Ha Doublet Coupled to H Doublet of Doublets

Hc coupled to Hd Doublet Coupled to H Doublet of Doublets

Hd coupled to Hc Doublet Coupled to H Doublet of Doublets

Multiplicity

Relative Intensitity

Remarks

Ha = Doublet of Doublets

Coupled to Hb (Doublet) Coupled to H (Doublet of Doublets)

Hb = Doublet of Doublets

Coupled to Ha (Doublet) Coupled to H (Doublet of Doublets)

Hc = Doublet of Doublets

Coupled to Hd (Doublet) Coupled to H (Doublet of Doublets)

Hd = Doublet of Doublets

Coupled to Hc (Doublet) Coupled to H (Doublet of Doublets)

H = Multiplet He = Triplet Hf = Triplet

1 2 2

Coupled to Ha , Hb , Hc , Hd Coupled to Hf Coupled to He

7 5 (a). Draw and indicate the relative intensities of 1H multiplets for FCH2OCH2CH2F by using the following typical values of coupling constants: 3JHH = 7 Hz, 2JHF = 60 Hz and 3JHF = 20 Hz.

8 5 (b). The chemical shift position for the 1H resonance in benzene (C6H6) is 7.27. How many hertz is this from the TMS resonance when the instrument is working at (i) and (ii)? (i) 200 MHz (ii) 400 MHz Chemical shift (ppm) = [ x Frequency of NMR instrument ] ppm (i) 200 MHz 7.27 ppm = [ x 200 MHz ] ppm 7.27 ppm = [ x 200 MHz ] ppm x = 200 MHz 7.27 = 1454 MHz = 1.454 109 Hz (ii) 400 MHz 7.27 ppm = [ x 400 MHz ] ppm 7.27 ppm = [ x 400 MHz ] ppm x = 400 MHz 7.27 = 2908 MHz = 2.908 109 Hz

6. One of the isomers of C3H6ClNO2, chloronitropropane, has a proton NMR spectrum consisting of a triplet ( 5.8), an approximate quintet ( 2.3) and a triplet ( 1.1). What isomer is this? Parent unit = Propane Direct to oxygen = 4 ppm Presence of NO2 = Pulls the chemical shift to even lower field = 5.8 ppm In presence of a chiral center, C*, the type of proton can/cannot be distinguished, depending on the instrument used: 200 MHz: Cannot distinguish the difference between Hb and Hb, thus, only can see Hb 600 MHz: Can distinguish the difference between Hb and Hb, thus, there will be Hb and Hb ~ Higher resolution can see more complicated structure(s)

quintet ( 2.3) triplet ( 5.8) Ha Hb Hc HaCCC*NO2 Ha Hb Cl triplet ( 1.1) Structure of 1-chloro-1-nitropropane (Isomer of chloronitropropane, C3H6ClNO2)

The signals in the spectrum can be readily assigned: = 5.8 ppm is due to the CH group adjacent to NO2 and Cl; = 2.3 ppm is due to the remaining CH2 group; = 1.1 ppm is due to the CH3 group. The multiplicity of the resonance at = 5.8 ppm is a triplet because it is coupled to 2 protons on the adjacent carbon. The multiplicity of the resonance at = 2.3 ppm is a quintet** because it is coupled to 1 proton at = 5.8 ppm and the 3 protons at = 1.1 ppm. The multiplicity of the resonance at = 1.1 ppm is a triplet because it is coupled to 2 protons on the adjacent carbon. ** quintet: n + 1 = 4 + 1 = 5 7. (a) Why NMR spectroscopy is said to be an insensitive technique compared to IR or UV spectroscopy? Compared with IR spectroscopy, the main problems in NMR experiments are lower sensitivity and resonance overlapping caused by large amounts of background chemicals present in the sample. Besides that, NMR requires milligrams (103 gram) of a typical organic molecule as compared to techniques like mass spectrometry require only nanograms (10 9 gram) of sample. This insensitivity stems primarily from the fact that only the difference in population at thermal equilibrium is active in the experiment. That means that only approximately one spin in 106 is actually detected in order to generate the NMR signal. The sensitivity of NMR is inherently low because the energy difference between nuclear spin states (between the ground and excited spin states) is very small, in the

10 order of 10-5 of the thermal energy at room temperature. In addition, at room temperature the energy levels are nearly equally populated. The IR spectrum alone, obtained with one data point per wavenumber can add more than 4000 measurements to the few classically determined properties. Thus, the IR spectrum of a material is a powerful set of physical properties (transmission elements) available for the identification of an unknown compound. Another important aspect is the relative sensitivity of different nuclei: because of the inherent differences between different nuclei in the strength of the nuclear magnet (), the signal strength received can be very much weaker than a proton signal. Low sensitivity, is due to low natural abundance and low magnetogyric ratios, therefore long relaxation times. The low sensitivity of NMR experiments can be compensated in part by concentrating the NMR samples and by running long-term experiments. Shorter recycle delays can be used to give higher sensitivity (per unit time), by trading off some loss of signal per scan through saturation for running more scans per unit time. However, this can be at the expense of quantitation. Hence, to improve sensitivity, NMR is usually coupled with other techniques, with gas chromatography (GC) or liquid chromatography (LC), which results in hyphenated techniques, GC-NMR and LC-NMR respectively, to provide superior analytical power achieved by combining selectivity and sensitivity.

7(b). Provide two fundamental reasons that may justify the need to have (buy) a 600 MHz system (NMR spectrometer) instead of a 200 MHz system. Resolution and intensity are the two fundamental reasons that may justify the need to have (buy) a 600 MHz system (NMR spectrometer) instead of a 200 MHz system.

E 600 MHz Bigger E

200 MHz Smaller E

11 The stronger the magnet, the bigger the energy gap, hence, greater energy difference, E, bigger difference in the Boltzmann distribution and bigger electron distribution, which results in higher intensity. Resolution is an important advantage to stronger magnets. Technically, resolution is the width of an NMR line measured in hertz at one half the height of the peak. The peak width depends on the rate of decay of the FID, which is determined by the homogeneity of the magnetic field (shimming) and the inherent rate of decay of the net magnetization in the xy plane (determined by a relaxation parameter of that proton called T 2). Peaks that are overlapped on the 200-MHz spectrometer are resolved (separated by a region of baseline with no intensity) on the 600-MHz spectrometer. This is clearly illustrated in Figure 4, peak g, which at low field is spread out and overlapped with some peaks on its upfield (right-hand) side.

Figure 4 A 600 MHz system has a stronger magnet which gives better signal-to-noise ratio, thus better sensitivity. A higher magnetic field strength (600 MHz) gives an advantage in sensitivity because increasing the B0 field increases the population difference between the and states proportionately. This increases the net magnetization of the sample at equilibrium and thus increases the FID signal received after the pulse. Another factor enters in during the recording of the FID: The Larmor frequency, 0, is proportional to B0, so the nuclear magnets precess at a higher speed when the magnetic field is increased, thus, spinning the net magnetization faster will generate a bigger FID. In a 13C acquisition on a 200-MHz instrument, it would require 27 times as long as the same experiment on a 600-MHz instrument to achieve the same signal-to-noise ratio (600/200 to the 1.5 power, then squared because signal-to-noise ratio varies with the square root of the number of scans).

12 8. The 1H NMR spectrum of dipropylether, (CH3CH2CH2)2O, is given below. The spectrum consists of 3 distinct resonances at 3.4 (triplet, 2H), 1.6 (multiplet, 2H) and 1.0 (triplet, 3H). Assign the spectrum and clearly describe the spectra that would be obtained while applying a strong Rf field at: (i) = 3.4 ppm Question 8 Question for Final Exam (ii) = 1.6 ppm (iii) = 1.0 ppm

The signals in the spectrum can be readily assigned ( = 3.4 ppm is due to the CH2 group adjacent to the oxygen; = 1.6 ppm is due to the remaining CH2 group and = 1.0 ppm is due to the CH3 group). The assignment is based on: (a) The relative intensities of the signals; (b) The observed splitting pattern; (c) The fact that the CH2 group adjacent to the oxygen must be the low field resonance. The multiplicity of the resonance at = 3.4 ppm is a triplet because it is coupled to 2 protons on the adjacent carbon. The multiplicity of the resonance at = 1.6 ppm is a triplet of quartets because it is coupled to the 2 protons at = 3.4 ppm (triplet splitting) and the 3 protons at = 1.0 ppm (quartet splitting). The effect of irradiation at each of these frequencies is to decouple the nuclei which have signals at these frequencies. Decoupling effectively removes the nucleus from the spin system. (i) With irradiation at = 3.4 ppm, the multiplicity of the CH2 resonance at = 1.6 ppm is simplified from a triplet of quartets to a quartet. The CH 3 resonance at = 1.0 ppm remains unchanged. (ii) With irradiation at = 1.6 ppm, the multiplicity of the CH 2 resonance at = 3.4 ppm is simplified from a triplet to a singlet. The CH3 resonance at = 1.0 ppm is simplified from a triplet to a singlet. (iii)With irradiation at = 1.0 ppm, the multiplicity of the CH 2 resonance at = 1.6 ppm is simplified from a triplet of quartets to a triplet. The CH 2 resonance at = 3.4 ppm remains unchanged.