Appendix VI - 1
May 2017
Appendix VI: DEPT, COSY and HETCOR (HMQC, HSQC & HMBC) NMR Spectra.
Explanation of DEPT spectra:
DEPT is an acronym for Distortionless Enhancement by Polarization Transfer. The result of this
experiment is a carbon spectrum that shows the multiplicities (methyl, methylene, methine and
quaternary) for the different carbons. DEPT is a 1D experiment, but unlike decoupling and NOE, it
involves a multiple pulse sequence. The experiment also requires that both protons and carbons are
excited by the pulses. The techniques and theory behind the DEPT experiment are very complicated,
but the results are easily interpreted.
There are three types of DEPT spectra that can be acquired. They are noted by numbers. DEPT-45
will give a positive1 signal for any carbon with an attached proton. DEPT-90 spectra will give a
positive signal for methine carbons. DEPT-135 spectra will give positive signals for methine and
methyl carbons, and negative signals for methylene carbons. By a linear combination of these three
spectra and the normal 13C spectrum, the multiplicities of all of the carbons in a molecule may be
determined. The example spectra below show the appearance of each type of carbon in the different
DEPT spectra.
The 45o and 135o subspectra contain peaks associated with carbons bearing protons (no quaternary
carbons visible). They differ only by the phase of the CH2 peaks (up and down respectively).
Thus, 45o subspectrum - 135o subspectrum = a subspectrum of just CH2 peaks
and 45o subspectrum + 135o subspectrum = a subspectrum of CH and CH3 peaks
The 90o subspectrum contains only the CH peaks, so subtracting this from the 45o + 135o
combination will give only the CH3 peaks.
1
The descriptors positive and negative have been used, but in fact the spectrum may appear either way up i.e.
the positive direction may be pointing either up or down.
Appendix VI - 2
In many cases, there is not much confusion between the chemical shifts of CH and CH3 protons, so
the 135o subspectrum effectively discriminates between each type of carbon.
COSY NMR Spectra
It is not intended that you tackle the theory behind two dimensional nmr spectra at this stage, but
there is no reason why you cannot use the results of the technique! If a spectrum contains coupled
resonances that are close together, the signals invariably become muddled to the point of not
providing any useful information. In which case, a COSY spectrum may well provide the answer.
Below is a brief summary of what happens in this type of experiment.
The regular pulse procedure for acquiring a spectrum is to have three different time intervals in each
cycle of scans. The first interval is when nothing happens and the spins are allowed to settle to their
natural distribution of states. The second is when a pulse of excitation energy is applied and the
spins respond to the stimulus. The detector is then switched on and measurement is taken for a time
t. The spectrum is obtained by transforming the data with respect to time t.
In a 2D experiment, a series of spectra are collected where the time between stimulus and detection
(t1) is increased by a small amount in each case. So not only can the spectra be transformed with
respect to the detection time (t2) but also to t1. This results in a contour plot with the same (but rather
crudely resolved) spectrum along each of two axes and a diagonal (bottom left to top right usually)
of the contours of the regular spectrum. The interest lies in the off-diagonal contours. Where
correlation between coupled
resonances exists, a contour
should appear in both halves of
the spectrum. Values of
coupling constants cannot be
determined by this method
(unless certain conditions are
applied) but the fact that two
resonances are coupled can aid in
the interpretation of the
spectrum.
Figure 2 shows a COSY
spect ru m of 1-propanol
containing three resonances. One
is coupled to the other two, but
they are not coupled to each
other. This is reflected in the size
of the contours (ie how big the
blobs are!).
Figure 2: COSY of 1-propanol
Appendix VI - 3
C-H Correlated Spectra including HMQC, HSQC & HMBC
Essentially, this type of spectrum is the same as a COSY, with the correlation between two different
nuclei. This means that there is no diagonal, but the correlation is signified by intensity at the cross
point. Figure 3 shows the HETCOR of 1-propanol.
Figure 3: HETCOR of 1-propanol
Modern instruments do the same experiment but using different pulse programs. HMQC and HSQC
both look at the correlation over 1J (ie direct coupling). If correlations over several bonds must be
observed, a filter is used to remove all 1J coupling of J>10 Hz. This is known as HMBC, or long-
range HETCOR.
References:
Derome, A.E. Modern NMR Techniques for Chemistry Research, Pergamon: Oxford, 1987.
Richards, S.A. Laboratory Guide to Proton NMR Spectroscopy, Blackwell: Oxford, 1988.
Keeler, J. Chem. Soc. Rev., 1990, 19, 381.