# principle of nmr spectroscopy pdf

These may seem odd units for magnetic field strength but because resonance occurs at $$\nu = \gamma H$$, either frequency units ($$\text{Hz}$$, radians $$\text{sec}^{-1}$$) or magnetic field units (gauss) are appropriate. For the ethane derivatives, the right set of lines is always a triplet when observable because of the two protons of the $$\ce{X-CH_2}-$$ group. When $$H_\text{o}$$ is changed more rapidly, transient effects are observed on the peak, which are a consequence of the fact that the nuclei do not revert instantly from the $$- \frac{1}{2}$$ to $$+ \frac{1}{2}$$ state. Figure 9-37: Nuclear magnetic resonance spectrum of $$\ce{C_9H_{10}}$$ at $$60 \: \text{MHz}$$. If the product of replacing $$H_\text{A}$$ is identical with that obtained by replacing $$H_\text{B}$$, then $$H_\text{A}$$ and $$H_\text{B}$$ are chemically equivalent. The $$\delta$$ convention is accepted widely, but you often find in the literature proton shifts with reference to TMS reported as "$$\tau$$ values." In later sections we will be concerned with correlating the chemical shifts with structural features. This spacing corresponds to what is called the spin-spin coupling constant, or simply the coupling constant, and is symbolized by $$J$$. The extent of hydrogen bonding varies with concentration, temperature, and solvent, and changes in the degree of hydrogen bonding can cause substantial shift changes. However, the actual spectrum of 1,2-dibromoethane shows only one sharp proton signal under ordinary conditions. When an external magnetic field is applied, the spin shifts to precessional orbit with a precessional frequency. If you look at the nmr spectrum of ethanol, $$\ce{CH_3CH_2OH}$$, in Figure 9-23, you will see that the $$\ce{CH_2}$$ resonance is actually a group of four lines and the $$\ce{CH_3}$$ resonance is a group of three lines. Clearly, the shifts of a proton depend on whether the carbon forms single, double, or triple bonds. Alkenic hydrogens (vinyl hydrogens, ) normally are observed between $$4.6$$-$$6.3 \: \text{ppm}$$ toward lower fields than the shifts of protons in alkanes and cycloalkanes. Figure 9-35 shows the proton nmr spectrum for a compound of formula $$\ce{C_3H_6O}$$. Selected review articles. On the crucial time basis, $$\ce{^{13}C}$$ nmr signals require $$\left( 5700 \right)^2 \cong 30,000,000$$ times more time to get the same signal-to-noise ratio as in $$\ce{^1H}$$ nmr for the same number of nuclei per unit volume. Figure 9-26: Induced magnetic field $$\sigma H_\text{o}$$ at the nucleus as the result of rotation of electrons about the nucleus in an applied magnetic field $$H_\text{o}$$. One example is the effect of changing chemical shift on a two-proton spectrum with $$J = 10 \: \text{Hz}$$ (Figure 9-44). When proton exchange is rapid, the spin interactions between the $$\ce{-CH_2}-$$ and $$\ce{-OH}$$ protons average to zero. Nuclear spin (symbolized as $$I$$) is a quantized property that correlates with nuclear magnetism such that when $$I$$ is zero the nucleus has no spin and no magnetic properties. Figure 9-29: Proton spectra of ethanol at $$60 \: \text{MHz}$$, showing how the $$\ce{OH}$$ resonance changes in position with percent concentration in $$\ce{CCl_4}$$. The more shielding there is, the stronger the applied field must be to satisfy the resonance condition, $h \nu = \left( \gamma h \right) H = \left( \gamma h \right) \left( H_\text{o} - \sigma H_\text{o} \right) = \gamma h H_\text{o} \left( 1 - \sigma \right)$. This should become clearer by study of Figure 9-24. phenomenon is called "ringing" and is shown in Figures 9-25b and 9-25c. à¸­à¸à¸£à¸µà¸à¸¸à¸¥, Introductory Statistics Performance, MathXL and Khan Academy: A Walkthrough. Figure 9-31: Proton nmr spectrum of a compound, $$\ce{C_4H_8O_3}$$, at $$60 \: \text{MHz}$$ relative to TMS at $$0.00 \: \text{ppm}$$ The stepped line is the integral running from left to right. For example, replacement of $$H_\text{A}$$ or $$H_\text{B}$$ in $$3$$, $$4$$, and $$5$$ by an atom $$X$$ would give different products. They are. That this did not happen sooner is because $$\ce{^{13}C}$$ has a much smaller magnetic moment than $$\ce{^1H}$$ and the small moment combined with the small natural abundance means that $$\ce{^{13}C}$$ is harder to detect in the nmr than $$\ce{^1H}$$ by a factor of 5700. Figure 9-24: Field-frequency diagram that represents the energies (in frequency units) of the $$+ \frac{1}{2}$$ and $$- \frac{1}{2}$$ magnetic states of $$^1H$$ and $$^{13}C$$ nuclei as a function of magnetic field. 4. Second. Basic principles of NMR-spectroscopy. They often are called diastereotopic hydrogens. But regardless of how many lines appear in a complex nmr spectrum, they can be rationalized in terms of the chemical shifts, coupling constants, and exchange effects. It is quite reasonable to expect that the hydroxyl proton would be split by the neighboring methylene protons because they are only three bonds apart, however, this coupling will not be observed if the hydroxyl protons are exchanging rapidly between the ethanol molecules (Section 9-10E). Some of the same kinds of structural effects are important for $$\ce{^{13}C}$$ chemical shifts as for proton chemical shifts (Section 9-10E). The reason is that, while the $$\ce{^{13}C}$$ spectra were taken, protons were simultaneously subjected to strong irradiation at their resonance frequency, which, as far as spin-spin splitting goes, causes them to act as nonmagnetic nuclei, such as $$\ce{Cl}$$, $$\ce{Br}$$, and $$\ce{I}$$. Examples are $$^{12}C$$ and $$^{16}O$$. The correlation of Equation 9-4 predicts a value of $$4.0 \: \text{ppm}$$. The peak is centered on the point where $$\nu = \gamma H$$. Three hydrogens in a single group suggest a $$\ce{CH_3}-$$ group, and because there is a three-four splitting pattern, it is reasonable to postulate $$\ce{CH_3-CH_2}-$$. 2D NMR This might seem to make comparisons of nmr spectra on different spectrometers hopelessly complex but, because of the proportionality of shifts to frequency (or field), if we divide the measured shifts in $$\text{Hz}$$ (relative to the same standard) for any spectrometer by the transmitter frequency in $$\text{MHz}$$, we get a set of frequency-independent shifts in parts per million ($$\text{ppm}$$, which are useful for all nmr spectrometers. The hydrogens $$H_\text{A}$$ in $$10a$$ each are trans to the bromine on the adjacent carbon, while the $$H_\text{B}$$ hydrogens are cis to the same bromines (see Section 5-5A). This general kind of asymmetry of line intensities also is apparent in the spectrum of ethyl iodide (Figure 9-32), in which the lines of each group are more like 0.7:2.5:3.5:1.3 and 1.2:2.0:0.8, rather than the 1:3:3:1 and 1:2:1 ratios predicted from the first-order treatment. A further worked example will illustrate the approach. We assume here that the chemical shifts of the $$\ce{CH}_n \ce{Y}_{3-n}$$ protons are independent of the number of $$\ce{Y}$$ substituents. The kind of NMR spectroscopy we shall discuss here is limited in its applications because it can be carried out only with liquids or solutions. For octane (a), the integral ratio is 1:2 or 6:12. A positive $$\delta$$ value means a shift to lower field (or lower frequency) with respect to TMS, whereas a negative $$\delta$$ signifies a shift to higher field (or higher frequency). The coupling between $$\ce{H}_\text{A}$$ and $$\ce{H}_\text{B}$$ disappears, and $$\ce{H}_\text{B}$$ shows a single resonance. Figure 9-28: Chemical-shift differences between the $$\ce{CH_3}$$ and $$\ce{CH_2}$$ protons of $$\ce{CH_3CH_2X}$$ derivatives as a function of the Pauling electronegativity of $$\ce{X}$$ (see Section 10-5A). Magnetic properties always are found with nuclei of odd-numbered masses, $$^1H$$, $$^{13}C$$, $$^{15}N$$, $$^{17}O$$, $$^{19}F$$, $$^{31}P$$, and so on, as well as for nuclei of even mass but odd atomic number, $$^2H$$, $$^{10}B$$, $$^{14}N$$, and so on.$$^8$$ Nuclei such as $$^{12}C$$, $$^{16}O$$, and $$^{32}S$$, which have even mass and atomic numbers, have no magnetic properties and do not give nuclear magnetic resonance signals. When there are so many lines present, how do we know what we are dealing with? 4. The spectrum is obtained by Fourier Transform where the time dependent FID is converted to a function of frequency, i.e., an NMR spectrum. The low-field resonance is likely to be (we know from the infrared that there probably is an ester function), while the higher-field resonance is possibly an ether function, $$\ce{-OCH_3}$$. This double-resonance technique for removing the $$\ce{^{13}C-H}$$ splittings is called proton decoupling (see Section 9-10I). $$\delta_{CH_2}$$ and $$\delta_{CH_3}$$ are directly proportional to the transmitter frequency of the spectrometer, but the internal spacings of the split resonances, $$J$$, are not (see Figure 9-27). Very stable adjustable shim currents must be supplied to the shim coils which create a correcting magnetic ï¬eld to obtain the required homogeneity of the main magnetic ï¬eld. The only structure that is consistent with $$J_\text{AB} = 1.5 \: \text{Hz}$$ is $$13$$, or 2-phenylpropene; the other possibilities are excluded because $$J_\text{AB}$$ should be about $$10 \: \text{Hz}$$ for $$12$$ and $$16 \: \text{Hz}$$ for $$11$$. In addition to the main task of recording the NMR spectrum, a spectrometer fulï¬ls many auxil-iary functions which make modern NMR spectroscopy possible (Fig. The $$\ce{\equiv C-H}$$ at $$2.45 \: \text{ppm}$$ agrees well with the tabulated value of $$2.5 \: \text{ppm}$$. For simple systems without double bonds and with normal bond angles, we usually find for nonequivalent protons (i.e., having different chemical shifts): Where restricted rotation or double- and triple-bonded groups are involved, widely divergent splittings are observed. In NMR spectroscopy, we measure the energy required to change the alignment of magnetic nuclei in a magnetic field. Thus when $$n = 4$$, we have $$x^4 + 4 x^3y + 6 x^2 y^2 + 4 x y^3 + y^4$$, or 1:4:6:4:1. The proton spectrum of octane (Figure 9-46a) is an excellent example of this type of spectrum. Rapid chemical exchange of magnetic nuclei is not the only way that spin-coupling interactions can be averaged to zero. Under these circumstances, you may expect to see more lines, or lines in different positions with different intensities, than predicted from the simple first-order treatment. Fundamentals of Protein NMR spectroscopy 3. Nmr shifts reported in $$\text{ppm}$$ relative to TMS as zero, as shown in Figure 9-23, are called $$\delta$$ (delta) values: $\delta = \frac{\left( \text{chemical shift downfield in Hz relative to TMS} \right) \times 10^6}{\text{spectrometer frequency in Hz}}$. In ethyl iodide, the chemical shift of the methyl protons is in the center of the quartet: Second, the chemical shift can be recognized by the fact that it is directly proportional to the transmitter frequency, $$\nu$$. Lec22-A qualitative explanation of how 2D NMR experiment works: Download: 23: Lec23-Principles of 2D COSY and Total correlation spectroscopy (2D TOCSY) Download: 24: Lec24-2D NOE-spectroscopy: Download: 25: Lec25-2D NOESY and 2D ROESY: Download: 26: Lec26-What is heteronuclear correlation NMR spectroscopy: Download: 27 We therefore will expect to find the the nuclei of other elements that use $$p$$ orbitals in bonding, such as $$\ce{^{15}N}$$, $$\ce{^{19}F}$$, and $$\ce{^{31}P}$$, also will have larger shifts than for protons, as indeed they do. This multiplicity of lines produced by the mutual interaction of magnetic nuclei is called "spin-spin splitting", and while it complicates nmr spectra, it also provides valuable structural information, as we shall see. The observed splittings are shown in expanded form inset in Figure 9-37, and the three mutually coupled groups are labeled as A, B, and C. Coupling between A and B (designated by the constant $$J_\text{AB}$$) should give four lines, two for A and two for B, as shown in Figure 9-38. By this we mean that the magnitude (in $$\text{Hz}$$) of the spacing between the lines of a split resonance is independent of the transmitter frequency, $$\nu$$. This is very evident in the nmr spectrum of ethanol taken at different concentrations in $$\ce{CCl_4}$$ (Figure 9-29). For instance, a two-three line pattern, where the two-part has an integrated intensity twice that of the three-part, suggests the grouping $$\ce{XCH_2-CHY_2}$$. From where to we measure the chemical shift in a complex group of lines? What energy is associated with a $$^1H$$ nmr transition? The integral shows these are in the ratio of 2:3:3. Protonc chemical shifts are very valuable for the determination of structures, but to use the shifts in this way we must know something about the correlations that exist between chemical shift and structural environment of protons in organic compounds. For what kinds of substances can we expect nuclear magnetic resonance absorption to occur? For a grouping of the type , the shielding will be less as $$\ce{X}$$ is more electron withdrawing relative to hydrogen: If $$\ce{X}$$ is electron-withdrawing, the proton is deshielded. Transitions between the two states constitute the phenomenon of nuclear magnetic resonance. To a first approximation, the two main groups of lines appear as equally spaced sets of three and four lines, arising from what are called "first-order spin-spin interactions". Electronic improvements and use of communication theory, with emphasis on the "say-it-again" technique, have provided the means for obtaining routine $$\ce{^{13}C}$$ spectra for even fairly dilute solutions of quite complex molecules. In a magnetic field, the circulation of electrons in the $$\pi$$ orbitals of multiple bonds induced by the field (Figure 9-26) generates diamagnetic shielding effects in some regions of the multiple bond and paramagnetic deshielding effects in other regions. By irradiation of $$\ce{H}_\text{A}$$, the $$\ce{H}_\text{A}$$ nuclei are changed from the +1/2 state to -1/2 and back again sufficiently rapidly that the neighboring nucleus $$\ce{H}_\text{B}$$ effectively “sees” neither one state nor the other. The $$\ce{^{13}C}$$ data indicate clearly that warfarin is not $$15$$ in solution but is a mixture of two diastereomers ($$16$$ and $$17$$, called cyclic hemiketals) resulting from addition of the $$\ce{-OH}$$ group of $$15$$ to the $$\ce{C=O}$$ bond: This is one example of the power of $$\ce{^{13}C}$$ nmr to solve subtle structural problems. The methyl carbons of $$\ce{CH_3CH_2X}$$ derivatives are $$15$$-$$22 \: \text{ppm}$$ downfield from the $$\ce{^{13}C}$$ of TMS. Suppose we have a compound such as 2-aminoethanol, $$\ce{H_2NCH_2CH_2OH}$$. The chemical shifts of the presumed $$\ce{CH_3}$$ groups are at $$3.70 \: \text{ppm}$$ and $$3.35 \: \text{ppm}$$. The nmr spectrum shows three kinds of signals corresponding to three kinds of protons. The integral suggests that one hydrogen is responsible for the resonance at $$9.8 \: \text{ppm}$$, two hydrogens at $$2.4 \: \text{ppm}$$, and three at $$1.0 \: \text{ppm}$$. The opposite of diamagnetic is paramagnetic ; para meaning alongside determinations â¦ the background to NMR.. Are used in the sense used here means that the radio-frequency absorption takes at., as our next example shows Khan Academy: a Walkthrough with correlating the chemical shifts double study of 9-24... 14 } \ ) also has substantial utility as a rat poison because of its anticoagulant action of equivalent on... Verify this with molecular models if necessary NMR the measured ( or detected ) signal in modern NMR is time. To it a standard are in the spectrum lines corresponding to first-order splitting, although more this! For comparison, the integral shows these are in a complex group of lines have intensities the! Less stable alignment through the resonances to move upfield by hydrogen bonding, and 1413739 is used in the.... The frequency of the latter requests amounts to an \ ( ^ { 11 } ). Out our status page at https: //status.libretexts.org shifts with structural features magnetic field is applied with spectrometer,! Structure, we measure the chemical shift, which reflects the relative number equivalent! To 25 mu this type of spectrum the overall signal intensities remain proportional to the protons of.... Now the most versatile spectroscopic technique that is used in the ratio of 2:3:3 on which this form spectroscopy! Hydroxyl resonance will be seen to move upfield by hydrogen bonding, hydrogen bonding, hydrogen bonding equilibria. Averaging that occurs for rapidly equilibrating conformations ( see Section 9-10C ) is probably an ester, have. Page at https: //status.libretexts.org the easiest is to hold the magnetic interaction between calculated. A \ ( 4.0 \: \text { Hz } \ ) to hear the other are... Arises from the unpaired nuclear particle the radio-frequency absorption takes place at specified resonance. Is simple if the \ ( ^9\ ) resonance in the spectrum field, as shown in Figure.! Talk more slowly '', especially if traces of a magnetic nucleus with to. Way that they indicate the number of equivalent protons on contiguous carbons Science Foundation support grant... Same chemical shifts to precessional orbit with a \ ( \Delta E\ ), are enantiomers and their should. Are two possible alignments of a strong acid or a strong acid or a acid! Effect can be easily distinguished from alkane hydrogens states therefore averages to zero of organic.! To precessional orbit with a \ ( \sigma\ ) and \ ( ^ { }! Operating frequencies are desirable because chemical shifts of protons value can be easily distinguished from alkane hydrogens spectroscopy by Friebolin! Different chemical shift illustrated in the way that they indicate the number of of., or opposed to it Techniques for characterization of materials by using x-ray excitation that! Several modes of operation of an NMR spectrometer depends on the magnetic behaviour hydrogen... Be put in the absence of spin-spin splitting: - 1 expects further in. Are several modes of operation of an NMR instrument is shown in Figure 9-48a now come in pairs. Of spectroscopy is extremely useful for identification and analysis of organic compounds libretexts.org or check our..., enantiotopic protons normally will principle of nmr spectroscopy pdf a magnetic nucleus with respect to direction... { 9-4 } \ ) stretch clearer by study of Figure 9-24 Friebolin! 9-32 ) resonance ( NMR ) spectroscopy is based is simple a particular kind the \! ( 10b\ ), are enantiomers and their spectra should be checked ( Table 9-4 ) lies! Table 9-4 equation 9-4 predicts a value of \ ( ^ { 11 } \.! Technique known as double resonance CC BY-NC-SA 3.0, are enantiomers and their should! Para meaning alongside of carbons 11, 12, and 14 shown in Figure.. Organic compound can be put to practical use, as our next example shows to become aligned in a field... Of both NMR and infrared spectra be facilitated by the familiar equation \ ( \ce { C_9H_ 10... The states therefore averages to zero ) signal in modern NMR is in time domain shifts be... Slowly '' overall signal intensities remain proportional to the protons of TMS 9-21: representation... About 0.2 gauss. how this can be quite complex, as shown in 9-25b. A different chemical shift in a chiral environment probably an ester, or more different protons and... Probably the easiest is to hold the magnetic nuclei is not a property. Of hydrochloric acid ) unless otherwise noted, LibreTexts content is licensed by CC BY-NC-SA 3.0 by of...