8.1
Chemical Shift: Internal References and Solvent Effects
In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added,…
Precise measurement of the absolute absorption frequencies of nuclei in a sample is difficult. To overcome this, a standard internal reference compound is added, and the difference between their absorption frequencies is measured.
Internal reference compounds, like tetramethylsilane, or TMS, are chemically inert, soluble in NMR solvents, and easily removable.
With highly shielded methyl protons that yield an intense signal at a lower frequency than most organic molecules, TMS is a primary reference in proton, carbon, and silicon NMR spectroscopy.
If the reference compound is not inert, it is kept in a capillary tube within the NMR tube and termed an external reference.
Further, deuterated NMR solvents contain residual protons whose signal can also be used as a secondary reference.
The signal from deuterium itself can be used to monitor the instrument's magnetic field by a technique called locking.
The deuterium signal is constantly compared to a reference frequency and adjusted if there is any variation.
View the full transcript and gain access to JoVE Core videos
Q1: Why is tetramethylsilane used as an internal reference in NMR spectroscopy?
Tetramethylsilane (TMS) is the primary internal reference compound in proton, carbon, and silicon NMR spectroscopy because it is chemically inert, soluble in NMR solvents, and easily removable. Its highly shielded methyl protons produce an intense signal at a lower frequency than most organic molecules, making it ideal for standardizing measurements.
Q2: What is the purpose of measuring frequency differences between a reference compound and sample signals?
Precise measurement of absolute absorption frequencies of nuclei is difficult in NMR. By measuring the frequency difference between an internal reference compound and sample signals, chemists overcome this limitation and obtain reliable, standardized chemical shift values that are independent of the instrument's magnetic field strength.
Q3: When is an external reference used instead of an internal reference in NMR?
An external reference is used when a suitable inert internal reference compound is unavailable. The external reference is kept in a capillary tube within the NMR tube, allowing measurement of frequency differences without direct contact between the reference and sample, preventing unwanted chemical interactions.
Q4: How can deuterated NMR solvents serve as secondary references?
Deuterated NMR solvents such as CDCl3, D2O, and (CD3)2SO contain residual protons whose signals can be used as secondary references. These residual proton signals provide an alternative reference point when the primary internal reference is unavailable or unsuitable for a particular analysis.
Q5: What is deuterium locking and why is it important in NMR spectroscopy?
Deuterium locking is a technique that uses the deuterium signal to monitor and stabilize the instrument's magnetic field. The deuterium signal is constantly compared to a reference frequency and adjusted if any variation occurs, ensuring consistent field strength and reliable spectroscopic measurements throughout data acquisition.
Q6: How do solvent effects influence NMR reference selection?
Solvent choice affects reference selection because the reference compound must be soluble in the chosen NMR solvent. Deuterated solvents like CDCl3 and D2O are commonly used because they dissolve both samples and references effectively while their residual protons or deuterium signals provide reliable reference points for chemical shift measurements.
Q7: What properties make tetramethylsilane ideal for standardizing chemical shift measurements?
TMS possesses several ideal properties: it is chemically inert and does not react with samples, it is soluble in standard NMR solvents, it is easily removable after analysis, and its highly shielded methyl protons yield an intense signal at lower frequency than most organic molecules, providing a clear, reliable reference point.