Circular Dichroism (CD) Spectroscopy

An introduction to circular dichroism spectroscopy

Circular dichroism (CD) is the difference in the absorption of left-handed circularly polarised light (L-CPL) and right-handed circularly polarised light (R-CPL) and occurs when a molecule contains one or more chiral chromophores (light-absorbing groups).


Circular dichroism = ΔA(λ) = A(λ)LCPL - A(λ)RCPL, where λ is the wavelength


Circular dichroism (CD) spectroscopy is a spectroscopic technique where the CD of molecules is measured over a range of wavelengths. CD spectroscopy is used extensively to study chiral molecules of all types and sizes, but it is in the study of large biological molecules where it finds its most important applications. A primary use is in analysing the secondary structure or conformation of macromolecules, particularly proteins as secondary structure is sensitive to its environment, temperature or pH, circular dichroism can be used to observe how secondary structure changes with environmental conditions or on interaction with other molecules. Structural, kinetic and thermodynamic information about macromolecules can be derived from circular dichroism spectroscopy.

Measurements carried out in the visible and ultra-violet region of the electro-magnetic spectrum monitor electronic transitions, and, if the molecule under study contains chiral chromophores then one CPL state will be absorbed to a greater extent than the other and the CD signal over the corresponding wavelengths will be non-zero. A circular dichroism signal can be positive or negative, depending on whether L-CPL is absorbed to a greater extent than R-CPL (CD signal positive) or to a lesser extent (CD signal negative). An example circular dichroism spectrum of a sample with multiple CD peaks is shown below, demonstrating how CD varies as a function of wavelength, and that a CD spectrum may exhibit both positive and negative peaks. 


Circular dichroism spectrum of vitamin B12

Circular dichroism spectra are measured using a circular dichroism spectrometer, such as the Chirascan, which is a highly specialised derivative of an ordinary absorption spectrometer. CD spectrometers measure alternately the absorption of L- and R-CPL, usually at a frequency of 50kHz, and then calculate the circular dichroism signal.


> Tutorial 1 Understanding Circular Dichroism

The British Assessment Bureau