Carbon-13 nuclear magnetic resonance

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Carbon-13 (C13) nuclear magnetic resonance (most commonly known as carbon-13 NMR spectroscopy or 13C NMR spectroscopy or sometimes simply referred to as carbon NMR) is the application of nuclear magnetic resonance (NMR) spectroscopy to carbon. It is analogous to proton NMR ( 1
H
 NMR) and allows the identification of carbon atoms in an organic molecule just as proton NMR identifies hydrogen atoms. 13C NMR detects only the 13
C
 isotope. The main carbon isotope, 12
C
 does not produce an NMR signal. Although ca. 1 mln. times less sensitive than 1H NMR spectroscopy, 13C NMR spectroscopy is widely used for characterizing organic and organometallic compounds, primarily because 1H-decoupled 13C-NMR spectra are more simple, have a greater sensitivity to differences in the chemical structure, and, thus, are better suited for identifying molecules in complex mixtures. [1] At the same time, such spectra lack quantitative information about the atomic ratios of different types of carbon nuclei, because nuclear Overhauser effect used in 1H-decoupled 13C-NMR spectroscopy enhances the signals from carbon atoms with a larger number of hydrogen atoms attached to them more than from carbon atoms with a smaller number of H's, and because full relaxation of 13C nuclei is usually not attained (for the sake of reducing the experiment time), and the nuclei with shorter relaxation times produce more intense signals.

Contents

The major isotope of carbon, the 12C isotope, has a spin quantum number of zero and so is not magnetically active and therefore not detectable by NMR. 13C, with a spin quantum number of 1/2, is not abundant (1.1%), whereas other popular nuclei are 100% abundant, e.g. 1H, 19F, 31P.

Receptivity

13C NMR spectroscopy is much less sensitive (ca. by 4 orders of magnitude) to carbon than 1H NMR spectroscopy is to hydrogen, because of the lower abundance (1.1%) of 13C compared to 1H (>99%), and because of a lower(0.702 vs. 2.8) nuclear magnetic moment. Stated equivalently, the gyromagnetic ratio (6.728284 107 rad T−1 s−1) is only 1/4th that of 1H. [2]

On the other hand, the sensitivity of 13C NMR spectroscopy benefits to some extent from nuclear Overhauser effect, which enhances signal intensity for non-quaternary 13C atoms.

Chemical shifts

The disadvantages in "receptivity" are compensated by the high sensitivity of 13C NMR signals to the chemical environment of the nucleus, i.e. the chemical shift "dispersion" is great, covering nearly 250 ppm. This dispersion reflects the fact that non-1H nuclei are strongly influenced by excited states ("paramagnetic" contribution to shielding tensor. This paramagnetic contribution is unrelated to paramagnetism). [3] For example, most 1H NMR signals for most organic compounds are within 15 ppm.

The chemical shift reference standard for 13C is the carbons in tetramethylsilane (TMS), [4] whose chemical shift is set as 0.0 ppm at every temperature.

Carbon-13 nuclear magnetic resonance

Typical chemical shifts in 13C-NMR

Coupling constants

Homonuclear 13C-13C coupling is normally only observed in samples that are enriched with 13C. The range for one-bond 1J(13C,13C) is 50–130 Hz. Two-bond 2J(13C,13C) are near 10 Hz.

The trends in J(1H,13C) and J(13C,13C) are similar, except that J(1H,13C are smaller owing to the modest value of the 13C nuclear magnetic moment. Values for 1J(1H,13C) range from 125 to 250 Hz. Values for 2J(1H,13C) are near 5 Hz and often are negative.

Implementation

Sensitivity

As a consequence of low receptivity, 13C NMR spectroscopy suffers from complications not encountered in proton NMR spectroscopy. Many measures can be implemented to compensate for the low receptivity of this nucleus. For example, high field magnets with internal bores are capable of accepting larger sample tubes (typically 10 mm in diameter for 13C NMR versus 5 mm for 1H NMR). Relaxation reagents allow more rapid pulsing. [5] A typical relaxation agent is chromium(III) acetylacetonate. For a typical sample, recording a 13C NMR spectrum may require several hours, compared to 15–30 minutes for 1H NMR. The nuclear dipole is weaker, the difference in energy between alpha and beta states is one-quarter that of proton NMR, and the Boltzmann population difference is correspondingly less. [6] One final measure to compensate for low receptivity is isotopic enrichment.

Some NMR probes, called cryoprobes, offer 20x signal enhancement relative to ordinary NMR probes. In cryoprobes, the RF generating and receiving electronics are maintained at ~ 25K using helium gas, which greatly enhances their sensitivity. [7] The trade-off is that cryoprobes are costly.

Coupling modes

Another potential complication results from the presence of large one bond J-coupling constants between carbon and hydrogen (typically from 100 to 250 Hz). While potentially informative, these couplings can complicate the spectra and reduce sensitivity. For these reasons, 13C-NMR spectra are usually recorded with proton NMR decoupling. Couplings between carbons can be ignored due to the low natural abundance of 13C. Hence in contrast to typical proton NMR spectra, which show multiplets for each proton position, carbon NMR spectra show a single peak for each chemically non-equivalent carbon atom. [8]

In further contrast to 1H NMR, the intensities of the signals are often not proportional to the number of equivalent 13C atoms. Instead, signal intensity is strongly influenced by (and proportional to) the number of surrounding spins (typically 1H). Integrations are more quantitative if the delay times are long, i.e. if the delay times greatly exceed relaxation times.

The most common modes of recording 13C spectra are proton-noise decoupling (also known as noise-, proton-, or broadband- decoupling), off-resonance decoupling, and gated decoupling. These modes are meant to address the large J values for 13C - H (110–320 Hz), 13C - C - H (5–60 Hz), and 13C - C - C - H (5–25 Hz) which otherwise make completely proton coupled 13C spectra difficult to interpret. [9]

With proton-noise decoupling, in which most spectra are run, a noise decoupler strongly irradiates the sample with a broad (approximately 1000 Hz) range of radio frequencies covering the range (such as 100 MHz for a 23,486 gauss field) at which protons change their nuclear spin. The rapid changes in proton spin create an effective heteronuclear decoupling, increasing carbon signal strength on account of the nuclear Overhauser effect (NOE) and simplifying the spectrum so that each non-equivalent carbon produces a singlet peak.

Both the atoms, carbon and hydrogen exhibit spins and are NMR active. The nuclear Overhauser Effect is in general, showing up when one of two different types of atoms is irradiated while the NMR spectrum of the other type is determined. If the absorption intensities of the observed (i.e., non-irradiated) atom change, enhancement occurs. The effect can be either positive or negative, depending on which atom types are involved [10] .

The relative intensities are unreliable because some carbons have a larger spin-lattice relaxation time and others have weaker NOE enhancement. [9]

In gated decoupling, the noise decoupler is gated on early in the free induction delay but gated off for the pulse delay. This largely prevents NOE enhancement, allowing the strength of individual 13C peaks to be meaningfully compared by integration, at a cost of half to two-thirds of the overall sensitivity. [9]

With off-resonance decoupling, the noise decoupler irradiates the sample at 1000–2000 Hz upfield or 2000–3000 Hz downfield of the proton resonance frequency. This retains couplings between protons immediately adjacent to 13C atoms but most often removes the others, allowing narrow multiplets to be visualized with one extra peak per bound proton (unless bound methylene protons are non-equivalent, in which case a pair of doublets may be observed). [9]

Distortionless enhancement by polarization transfer spectra

Various DEPT spectra of propyl benzoate From top to bottom: 135deg, 90deg and 45deg DEPT spectra.jpg
Various DEPT spectra of propyl benzoate
From top to bottom: 135°, 90° and 45°

Distortionless enhancement by polarization transfer (DEPT) [11] is an NMR method used for determining the presence of primary, secondary and tertiary carbon atoms. The DEPT experiment differentiates between CH, CH2 and CH3 groups by variation of the selection angle parameter (the tip angle of the final 1H pulse): 135° angle gives all CH and CH3 in a phase opposite to CH2; 90° angle gives only CH groups, the others being suppressed; 45° angle gives all carbons with attached protons (regardless of number) in phase.

Signals from quaternary carbons and other carbons with no attached protons are always absent (due to the lack of attached protons).

The polarization transfer from 1H to 13C has the secondary advantage of increasing the sensitivity over the normal 13C spectrum (which has a modest enhancement from the nuclear overhauser effect (NOE) due to the 1H decoupling).

Attached proton test spectra

Another useful way of determining how many protons a carbon in a molecule is bonded to is to use an attached proton test (APT), which distinguishes between carbon atoms with even or odd number of attached hydrogens. A proper spin-echo sequence is able to distinguish between S, I2S and I1S, I3S spin systems: the first will appear as positive peaks in the spectrum, while the latter as negative peaks (pointing downwards), while retaining relative simplicity in the spectrum since it is still broadband proton decoupled.

Even though this technique does not distinguish fully between CHn groups, it is so easy and reliable that it is frequently employed as a first attempt to assign peaks in the spectrum and elucidate the structure. [12] Additionally, signals from quaternary carbons and other carbons with no attached protons are still detectable, so in many cases an additional conventional 13C spectrum is not required, which is an advantage over DEPT. It is, however, sometimes possible that a CH and CH2 signal have coincidentally equivalent chemical shifts resulting in annulment in the APT spectrum due to the opposite phases. For this reason the conventional 13C{1H} spectrum or HSQC are occasionally also acquired.

See also

Related Research Articles

The nuclear Overhauser effect (NOE) is the transfer of nuclear spin polarization from one population of spin-active nuclei to another via cross-relaxation. A phenomenological definition of the NOE in nuclear magnetic resonance spectroscopy (NMR) is the change in the integrated intensity of one NMR resonance that occurs when another is saturated by irradiation with an RF field. The change in resonance intensity of a nucleus is a consequence of the nucleus being close in space to those directly affected by the RF perturbation.

In nuclear magnetic resonance (NMR) spectroscopy, the chemical shift is the resonant frequency of an atomic nucleus relative to a standard in a magnetic field. Often the position and number of chemical shifts are diagnostic of the structure of a molecule. Chemical shifts are also used to describe signals in other forms of spectroscopy such as photoemission spectroscopy.

<span class="mw-page-title-main">Nuclear magnetic resonance spectroscopy</span> Laboratory technique

Nuclear magnetic resonance spectroscopy, most commonly known as NMR spectroscopy or magnetic resonance spectroscopy (MRS), is a spectroscopic technique based on re-orientation of atomic nuclei with non-zero nuclear spins in an external magnetic field. This re-orientation occurs with absorption of electromagnetic radiation in the radio frequency region from roughly 4 to 900 MHz, which depends on the isotopic nature of the nucleus and increased proportionally to the strength of the external magnetic field. Notably, the resonance frequency of each NMR-active nucleus depends on its chemical environment. As a result, NMR spectra provide information about individual functional groups present in the sample, as well as about connections between nearby nuclei in the same molecule. As the NMR spectra are unique or highly characteristic to individual compounds and functional groups, NMR spectroscopy is one of the most important methods to identify molecular structures, particularly of organic compounds.

<span class="mw-page-title-main">Solid-state nuclear magnetic resonance</span>

Solid-state NMR (ssNMR) spectroscopy is a technique for characterizing atomic level structure in solid materials e.g. powders, single crystals and amorphous samples and tissues using nuclear magnetic resonance (NMR) spectroscopy. The anisotropic part of many spin interactions are present in solid-state NMR, unlike in solution-state NMR where rapid tumbling motion averages out many of the spin interactions. As a result, solid-state NMR spectra are characterised by larger linewidths than in solution state NMR, which can be utilized to give quantitative information on the molecular structure, conformation and dynamics of the material. Solid-state NMR is often combined with magic angle spinning to remove anisotropic interactions and improve the resolution as well as the sensitivity of the technique.

<span class="mw-page-title-main">Proton nuclear magnetic resonance</span> NMR via protons, hydrogen-1 nuclei

Proton nuclear magnetic resonance is the application of nuclear magnetic resonance in NMR spectroscopy with respect to hydrogen-1 nuclei within the molecules of a substance, in order to determine the structure of its molecules. In samples where natural hydrogen (H) is used, practically all the hydrogen consists of the isotope 1H.

Nuclear magnetic resonance spectroscopy of proteins is a field of structural biology in which NMR spectroscopy is used to obtain information about the structure and dynamics of proteins, and also nucleic acids, and their complexes. The field was pioneered by Richard R. Ernst and Kurt Wüthrich at the ETH, and by Ad Bax, Marius Clore, Angela Gronenborn at the NIH, and Gerhard Wagner at Harvard University, among others. Structure determination by NMR spectroscopy usually consists of several phases, each using a separate set of highly specialized techniques. The sample is prepared, measurements are made, interpretive approaches are applied, and a structure is calculated and validated.

The heteronuclear single quantum coherence or heteronuclear single quantum correlation experiment, normally abbreviated as HSQC, is used frequently in NMR spectroscopy of organic molecules and is of particular significance in the field of protein NMR. The experiment was first described by Geoffrey Bodenhausen and D. J. Ruben in 1980. The resulting spectrum is two-dimensional (2D) with one axis for proton (1H) and the other for a heteronucleus, which is usually 13C or 15N. The spectrum contains a peak for each unique proton attached to the heteronucleus being considered. The 2D HSQC can also be combined with other experiments in higher-dimensional NMR experiments, such as NOESY-HSQC or TOCSY-HSQC.

Two-dimensional nuclear magnetic resonance spectroscopy is a set of nuclear magnetic resonance spectroscopy (NMR) methods which give data plotted in a space defined by two frequency axes rather than one. Types of 2D NMR include correlation spectroscopy (COSY), J-spectroscopy, exchange spectroscopy (EXSY), and nuclear Overhauser effect spectroscopy (NOESY). Two-dimensional NMR spectra provide more information about a molecule than one-dimensional NMR spectra and are especially useful in determining the structure of a molecule, particularly for molecules that are too complicated to work with using one-dimensional NMR.

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In nuclear chemistry and nuclear physics, J-couplings are mediated through chemical bonds connecting two spins. It is an indirect interaction between two nuclear spins that arises from hyperfine interactions between the nuclei and local electrons. In NMR spectroscopy, J-coupling contains information about relative bond distances and angles. Most importantly, J-coupling provides information on the connectivity of chemical bonds. It is responsible for the often complex splitting of resonance lines in the NMR spectra of fairly simple molecules.

Nuclear magnetic resonance (NMR) in the geomagnetic field is conventionally referred to as Earth's field NMR (EFNMR). EFNMR is a special case of low field NMR.

Carbon satellites in physics and spectroscopy, are small peaks that can be seen shouldering the main peaks in the nuclear magnetic resonance (NMR) spectrum. These peaks can occur in the NMR spectrum of any NMR active atom where those atoms adjoin a carbon atom. However, Carbon satellites are most often encountered in proton NMR.

<span class="mw-page-title-main">Phosphorus-31 nuclear magnetic resonance</span> Spectroscopy technique for molecules containing phosphorus

Phosphorus-31 NMR spectroscopy is an analytical chemistry technique that uses nuclear magnetic resonance (NMR) to study chemical compounds that contain phosphorus. Phosphorus is commonly found in organic compounds and coordination complexes, making it useful to measure 31- NMR spectra routinely. Solution 31P-NMR is one of the more routine NMR techniques because 31P has an isotopic abundance of 100% and a relatively high gyromagnetic ratio. The 31P nucleus also has a spin of 1/2, making spectra relatively easy to interpret. The only other highly sensitive NMR-active nuclei spin 1/2 that are monoisotopic are 1H and 19F.

<span class="mw-page-title-main">Fluorine-19 nuclear magnetic resonance spectroscopy</span> Analytical technique

Fluorine-19 nuclear magnetic resonance spectroscopy is an analytical technique used to detect and identify fluorine-containing compounds. 19F is an important nucleus for NMR spectroscopy because of its receptivity and large chemical shift dispersion, which is greater than that for proton nuclear magnetic resonance spectroscopy.

Carbohydrate NMR spectroscopy is the application of nuclear magnetic resonance (NMR) spectroscopy to structural and conformational analysis of carbohydrates. This method allows the scientists to elucidate structure of monosaccharides, oligosaccharides, polysaccharides, glycoconjugates and other carbohydrate derivatives from synthetic and natural sources. Among structural properties that could be determined by NMR are primary structure, saccharide conformation, stoichiometry of substituents, and ratio of individual saccharides in a mixture. Modern high field NMR instruments used for carbohydrate samples, typically 500 MHz or higher, are able to run a suite of 1D, 2D, and 3D experiments to determine a structure of carbohydrate compounds.

Nuclear magnetic resonance decoupling is a special method used in nuclear magnetic resonance (NMR) spectroscopy where a sample to be analyzed is irradiated at a certain frequency or frequency range to eliminate or partially the effect of coupling between certain nuclei. NMR coupling refers to the effect of nuclei on each other in atoms within a couple of bonds distance of each other in molecules. This effect causes NMR signals in a spectrum to be split into multiple peaks. Decoupling fully or partially eliminates splitting of the signal between the nuclei irradiated and other nuclei such as the nuclei being analyzed in a certain spectrum. NMR spectroscopy and sometimes decoupling can help determine structures of chemical compounds.

<span class="mw-page-title-main">Nuclear magnetic resonance</span> Spectroscopic technique based on change of nuclear spin state

Nuclear magnetic resonance (NMR) is a physical phenomenon in which nuclei in a strong constant magnetic field are perturbed by a weak oscillating magnetic field and respond by producing an electromagnetic signal with a frequency characteristic of the magnetic field at the nucleus. This process occurs near resonance, when the oscillation frequency matches the intrinsic frequency of the nuclei, which depends on the strength of the static magnetic field, the chemical environment, and the magnetic properties of the isotope involved; in practical applications with static magnetic fields up to ca. 20 tesla, the frequency is similar to VHF and UHF television broadcasts (60–1000 MHz). NMR results from specific magnetic properties of certain atomic nuclei. High-resolution nuclear magnetic resonance spectroscopy is widely used to determine the structure of organic molecules in solution and study molecular physics and crystals as well as non-crystalline materials. NMR is also routinely used in advanced medical imaging techniques, such as in magnetic resonance imaging (MRI). The original application of NMR to condensed matter physics is nowadays mostly devoted to strongly correlated electron systems. It reveals large many-body couplings by fast broadband detection and should not be confused with solid state NMR, which aims at removing the effect of the same couplings by Magic Angle Spinning techniques.

Nucleic acid NMR is the use of nuclear magnetic resonance spectroscopy to obtain information about the structure and dynamics of nucleic acid molecules, such as DNA or RNA. It is useful for molecules of up to 100 nucleotides, and as of 2003, nearly half of all known RNA structures had been determined by NMR spectroscopy.

Triple resonance experiments are a set of multi-dimensional nuclear magnetic resonance spectroscopy (NMR) experiments that link three types of atomic nuclei, most typically consisting of 1H, 15N and 13C. These experiments are often used to assign specific resonance signals to specific atoms in an isotopically-enriched protein. The technique was first described in papers by Ad Bax, Mitsuhiko Ikura and Lewis Kay in 1990, and further experiments were then added to the suite of experiments. Many of these experiments have since become the standard set of experiments used for sequential assignment of NMR resonances in the determination of protein structure by NMR. They are now an integral part of solution NMR study of proteins, and they may also be used in solid-state NMR.

Nitrogen-15 nuclear magnetic resonance spectroscopy is a version of nuclear magnetic resonance spectroscopy that examines samples containing the 15N nucleus. 15N NMR differs in several ways from the more common 13C and 1H NMR. To circumvent the difficulties associated with measurement of the quadrupolar, spin-1 14N nuclide, 15N NMR is employed in samples for detection since it has a ground-state spin of ½. Since14N is 99.64% abundant, incorporation of 15N into samples often requires novel synthetic techniques.

References

  1. ↑ Brian E. Mann, Brian F. Taylor (1981). ¹³C NMR data for organometallic compounds. Academic Press. ISBN   9780124691506.
  2. ↑ R. M. Silverstein; G. C. Bassler; T. C. Morrill (1991). Spectrometric Identification of Organic Compounds . Wiley. ISBN   9780471634041.
  3. ↑ Peter Atkins (2009). Physical Chemistry (5 ed.). Freeman.
  4. ↑ "The Theory of NMR - Chemical Shift". Archived from the original on 2015-01-23. Retrieved 2014-01-23.
  5. ↑ Caytan E, Remaud GS, Tenailleau E, Akoka S (2007). "Precise and Accurate Quantitative 13C NMR with Reduced Experimental Time". Talanta. 71 (3): 1016–1021. doi:10.1016/j.talanta.2006.05.075. PMID   19071407.
  6. ↑ "Measuring 13C NMR Spectra". University of Wisconsin.
  7. ↑ Molinski, Tadeusz F. (2010). "NMR of natural products at the 'nanomole-scale'". Natural Product Reports. 27 (3): 321–9. doi:10.1039/B920545B. PMID   20179874.
  8. ↑ "Introduction to Carbon NMR". University of Puget Sound.
  9. 1 2 3 4 Lal Dhar Singh Yadav (2013-08-13). Organic Spectroscopy. Springer. pp. 197–199. ISBN   9781402025754.
  10. ↑ Pavia, Donald L., ed. (2009). Introduction to spectroscopy (4th ed.). Belmont, CA: Brooks/Cole, Cengage Learning. ISBN   978-0-495-11478-9.
  11. ↑ Doddrell, D.M.; Pegg, D.T.; Bendall, M.R. (1982). "Distortionless enhancement of NMR signals by polarization transfer". J. Magn. Reson. 48 (2): 323–327. Bibcode:1982JMagR..48..323D. doi:10.1016/0022-2364(82)90286-4.
  12. ↑ Keeler, James (2010). Understanding NMR Spectroscopy (2nd ed.). John Wiley & Sons. p. 457. ISBN   978-0-470-74608-0.
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