🥼Organic Chemistry Unit 13 – NMR Spectroscopy for Structure Determination

Fundamentals of NMR Spectroscopy

• NMR spectroscopy utilizes the magnetic properties of certain atomic nuclei to determine the structure of molecules
• Relies on the principle of nuclear magnetic resonance, where nuclei absorb and re-emit electromagnetic radiation in a magnetic field
• Provides detailed information about the chemical environment of individual atoms within a molecule
• Non-destructive technique that allows for the analysis of pure compounds and mixtures without altering the sample
• Requires the presence of NMR-active nuclei, such as $^1$H, $^{13}$C, $^{19}$F, and $^{31}$P, in the molecule being analyzed
  • These nuclei possess a property called spin, which enables them to interact with external magnetic fields
• Magnetic field strength is measured in units of Tesla (T) or MHz, with higher field strengths providing better resolution and sensitivity
• Resulting NMR spectra display signals corresponding to different chemical environments of the NMR-active nuclei within the molecule

NMR-Active Nuclei and Their Properties

• NMR-active nuclei have a non-zero spin quantum number (I), which determines their magnetic properties
  • Nuclei with even mass numbers and even atomic numbers (e.g., $^{12}$C, $^{16}$O) have I = 0 and are NMR-inactive
  • Nuclei with odd mass numbers (e.g., $^1$H, $^{13}$C, $^{19}$F) have I = 1/2 and are the most commonly used in NMR spectroscopy
• The spin quantum number determines the number of possible orientations (2I + 1) of the nucleus in an external magnetic field
  • For I = 1/2 nuclei, there are two possible orientations: α (aligned with the field) and β (opposed to the field)
• The energy difference between the α and β states is proportional to the strength of the external magnetic field ($B_0$) and the gyromagnetic ratio (γ) of the nucleus
  • $ΔE = \frac{hγB_0}{2π}$, where h is Planck's constant
• The gyromagnetic ratio is a constant for each NMR-active nucleus and determines its sensitivity and resonance frequency
• The natural abundance of NMR-active isotopes varies, with $^1$H being the most abundant (99.98%) and $^{13}$C being relatively rare (1.11%)
  • This affects the sensitivity and acquisition time required for different nuclei

Chemical Shifts and Shielding Effects

• Chemical shift (δ) is a measure of the resonance frequency of a nucleus relative to a standard reference compound
  • Expressed in parts per million (ppm) and is independent of the magnetic field strength
  • $δ = \frac{ν_{sample} - ν_{reference}}{ν_{reference}} \times 10^6$, where ν is the resonance frequency
• The chemical shift of a nucleus is determined by its chemical environment, which is influenced by the electron density surrounding the nucleus
• Shielding effects occur when electrons circulate around the nucleus, generating a local magnetic field that opposes the external magnetic field
  • Increased electron density results in greater shielding and a lower (upfield) chemical shift
  • Decreased electron density leads to deshielding and a higher (downfield) chemical shift
• Factors that influence shielding include electronegativity of neighboring atoms, hybridization, and the presence of aromatic rings
  • Electronegative atoms (e.g., O, N, F) withdraw electron density and cause deshielding
  • sp$^3$ hybridized carbons are more shielded than sp$^2$ and sp hybridized carbons
  • Aromatic rings generate a ring current that shields nearby nuclei
• The chemical shift range for $^1$H NMR is typically 0-12 ppm, while $^{13}$C NMR has a wider range of 0-200 ppm

Spin-Spin Coupling and Multiplicity

• Spin-spin coupling occurs when the magnetic moments of neighboring NMR-active nuclei interact with each other
• Coupling between nuclei leads to the splitting of NMR signals into multiplets, with the number of peaks determined by the n+1 rule
  • n is the number of equivalent neighboring nuclei coupling to the observed nucleus
  • Multiplicity follows the pattern: singlet (n=0), doublet (n=1), triplet (n=2), quartet (n=3), etc.
• Coupling constants (J) measure the strength of the interaction between coupled nuclei and are expressed in Hertz (Hz)
  • J is independent of the magnetic field strength and can provide information about bond angles and stereochemistry
• First-order coupling occurs when the chemical shift difference between coupled nuclei is much larger than the coupling constant
  • Results in symmetrical multiplets with predictable peak intensities based on Pascal's triangle
• Second-order coupling arises when the chemical shift difference is comparable to the coupling constant
  • Leads to more complex, asymmetric multiplets and roofing effects
• Homonuclear coupling involves the interaction between the same type of nuclei (e.g., $^1$H-$^1$H), while heteronuclear coupling occurs between different types of nuclei (e.g., $^1$H-$^{13}$C)
• Coupling can be transmitted through bonds (scalar coupling) or through space (dipolar coupling)
  • Scalar coupling is more common and is used to determine the connectivity of atoms in a molecule

1D Proton NMR Spectra Interpretation

• 1D $^1$H NMR is the most common type of NMR experiment and provides a wealth of structural information
• The number of signals in a $^1$H NMR spectrum indicates the number of distinct chemical environments for hydrogen atoms in the molecule
• Signal integration measures the relative number of hydrogen atoms contributing to each peak
  • The area under each signal is proportional to the number of hydrogen atoms in that environment
  • Integration values are typically normalized to the smallest integer value
• Multiplicity of signals arises from spin-spin coupling and provides information about the number of neighboring hydrogen atoms
• Coupling constants can be measured as the distance between peaks in a multiplet and provide information about bond angles and stereochemistry
• The chemical shift of signals is influenced by the electron density around the hydrogen atoms and can be used to identify functional groups
  • For example, aldehydic protons appear around 9-10 ppm, aromatic protons around 6-8 ppm, and aliphatic protons around 0-4 ppm
• Exchangeable protons (e.g., -OH, -NH) may appear as broad signals or be absent due to rapid exchange with solvent molecules
• Interpretation of $^1$H NMR spectra involves correlating the observed signals with the structure of the molecule, considering the number of signals, integration, multiplicity, and chemical shifts

13C NMR Spectroscopy

• $^{13}$C NMR spectroscopy provides complementary information to $^1$H NMR and is particularly useful for determining the carbon skeleton of a molecule
• The low natural abundance of $^{13}$C (1.11%) results in lower sensitivity compared to $^1$H NMR
  • Requires longer acquisition times or higher sample concentrations to obtain good signal-to-noise ratios
• $^{13}$C NMR spectra typically display single peaks for each unique carbon environment, as the probability of two $^{13}$C nuclei being adjacent to each other is very low
• The chemical shift range for $^{13}$C NMR is much wider than for $^1$H NMR, spanning from 0-200 ppm
  • Allows for better resolution and differentiation of carbon environments
• Factors influencing $^{13}$C chemical shifts include hybridization, electronegativity of neighboring atoms, and the presence of multiple bonds
  • sp$^3$ hybridized carbons appear around 0-80 ppm, sp$^2$ carbons around 100-150 ppm, and sp carbons around 60-90 ppm
  • Electronegative substituents cause a downfield shift in the $^{13}$C signal
  • Double and triple bonds also result in downfield shifts
• Broadband decoupling is often employed in $^{13}$C NMR to simplify the spectra by removing the effects of $^1$H-$^{13}$C coupling
  • Results in singlets for each carbon environment, facilitating interpretation
• $^{13}$C NMR can be used to identify quaternary carbons, which are not directly observable in $^1$H NMR due to the absence of attached hydrogen atoms
• Comparison of $^{13}$C NMR chemical shifts with reference data and prediction tools can aid in structural elucidation

Advanced NMR Techniques

• 2D NMR experiments provide additional information about the connectivity and spatial relationships between nuclei in a molecule
  • Common 2D techniques include COSY, HSQC, HMBC, and NOESY
• COSY (COrrelation SpectroscopY) identifies scalar coupling between protons, helping to establish the connectivity of the carbon skeleton
  • Cross-peaks in a COSY spectrum indicate protons that are coupled to each other
• HSQC (Heteronuclear Single Quantum Coherence) correlates protons with directly attached carbons
  • Useful for assigning carbon signals and identifying the number of hydrogen atoms attached to each carbon
• HMBC (Heteronuclear Multiple Bond Correlation) detects long-range coupling between protons and carbons, typically 2-4 bonds apart
  • Helps to establish the connectivity of the molecule and identify quaternary carbons
• NOESY (Nuclear Overhauser Effect SpectroscopY) identifies protons that are spatially close to each other, even if not directly bonded
  • Useful for determining the relative stereochemistry and conformation of molecules
• DEPT (Distortionless Enhancement by Polarization Transfer) experiments selectively enhance the signals of CH, CH$_2$, and CH$_3$ groups in $^{13}$C NMR
  • Helps to identify the number of hydrogen atoms attached to each carbon
• Quantitative NMR (qNMR) techniques allow for the accurate determination of sample concentration and purity
  • Requires the use of an internal standard with a known concentration and a long relaxation delay to ensure complete relaxation of all nuclei
• Solid-state NMR spectroscopy enables the analysis of samples that are not soluble or are in the solid phase
  • Techniques such as magic angle spinning (MAS) and cross-polarization (CP) are employed to improve resolution and sensitivity

Solving Unknown Structures with NMR Data

• Determining the structure of an unknown compound using NMR data involves a systematic approach that combines information from various NMR experiments
• Begin by analyzing the 1D $^1$H NMR spectrum to identify the number of distinct hydrogen environments, their chemical shifts, multiplicities, and coupling constants
  • Use the integration values to determine the relative number of hydrogen atoms in each environment
• Compare the observed chemical shifts with reference data to identify potential functional groups and structural features
• Analyze the $^{13}$C NMR spectrum to determine the number of unique carbon environments and their chemical shifts
  • Use DEPT experiments to identify the number of hydrogen atoms attached to each carbon
• Combine the information from $^1$H and $^{13}$C NMR to construct fragments of the molecule and establish connectivity
• Use 2D NMR experiments to confirm the connectivity and spatial relationships between atoms
  • COSY and TOCSY help establish the proton-proton connectivity
  • HSQC correlates protons with directly attached carbons
  • HMBC identifies long-range proton-carbon correlations, helping to connect fragments and identify quaternary carbons
• NOESY experiments can provide information about the relative stereochemistry and conformation of the molecule
• Consider the molecular formula, which can be obtained from mass spectrometry or elemental analysis, to ensure that the proposed structure is consistent with the data
• Use NMR prediction software to compare the observed chemical shifts and coupling constants with those predicted for the proposed structure
• Iterate the process, refining the structure based on any inconsistencies or additional information until a final structure is determined that satisfies all the available data
• Confirm the structure through comparison with literature data, if available, or by synthesizing the compound and comparing its NMR spectra with the unknown sample