ChemActivity L2: Mass Spectrometry 283

The Big Picture

Mass spectroscopy is often the first tool in determining the identity of an unknown compound because
simple (low resolution) mass spectroscopy usually yields an accurate molecular formula.
More complex instruments can compare a spectrum to those of known molecules, giving a positive
identification of an exact structure. This is how drug testing is carried out. This spectrum matching is
similar to matching the fingerprint region of an IR spectrum, but much more accurate. One way this is
done is to match fragmentation patterns. That is, a molecule falls apart in the same way each time,
leaving telltale clues about its identity in the pattern of fragmentation peaks (peaks smaller than the
[M]+ peak).
Identification of fragmentation patterns can help you figure out the most likely structure or structures
among the many isomeric possibilities. There is a science to predicting fragmentation patterns, but that
topic is not covered in this activity. A simplistic (but often effective) way of predicting fragments is to
chop up the molecule near any functional group.

Common Points of Confusion

• Students commonly make the error of assuming that the base peak gives the molecular weight of
an unknown compound. The base peak is sometimes also the molecular ion, but more often these
peaks are not the same. The only time they are the same is when the molecular ion is the most
abundant ion detected, making the molecular ion peak the tallest peak in the spectrum.
• Do not make the mistake of assuming that a calculation of the number of carbon atoms based on
the height of the [M+1]+ peak is set in stone. Any calculation of the number of carbon atoms (or
especially the number of oxygen, sulfur or other atoms) is subject to a high degree of error due to
the usually small height of the [M+1]+ (or [M+2]+) peak. You should always consider the
possibility that such calculated numbers could be off by +/- 1.
ChemActivity L3: Carbon (13C) NMR Spectroscopy

BUILD MODELS: 1-bromo-2,3,3-trimethylbutane

(What can a 13C NMR spectrum tell you about the structure of a molecule?)
This activity is designed to be completed in a 1-½-hour laboratory session or two classroom sessions.

Model 1: Spectroscopy Using Radio Waves

In the presence of an external magnetic field, a 13C atom will absorb light in the radio frequency range.
The precise frequency of the light absorbed tells us about the atoms near that carbon atom.
For example: the five different carbon atoms labeled a to e below absorb light of different frequencies.
Frequency is measured along the x axis on the chart in funny units called ppm. Each cluster of peaks
(labeled with a letter) indicates a unique carbon that absorbs light at that frequency.
When the light is absorbed, it causes what is called a nuclear spin flip, so this type of spectroscopy is
called nuclear magnetic resonance (or NMR) spectroscopy.
NMR is the most widely used tool for determining the structure of molecules.
It turns out that the physics of NMR spectroscopy is quite complex. Since its discovery in the 1940s it has grown to become its
own subfield of chemistry. The simplified explanation above will be enough for us to learn to interpret NMR spectra.

d e each line is a very thin peak

H3C CH3 e
e a
b CH cC CH3
d
H2C a c
CH3 b
e
Br

baseline "noise"

100 80 60 40 20 0 ppm
13
Figure L3.1: Cartoon of (coupled) C NMR Spectrum of 1-bromo-2,3,3-trimethylbutane

Critical Thinking Questions

1. Next to each carbon in the structure, indicate the number of hydrogens attached to that carbon.
Also write this number next to the corresponding peak cluster on the spectrum.
2. Construct an explanation for why the three carbon atoms labeled “e” are called equivalent carbons
(or chemically indistinguishable carbons). A model may help. Note that the three carbons labeled
"e" are also called NMR equivalent. This is because the peak clusters due to the three “e” carbons
are on top of each other (appear as one large peak cluster) on the NMR spectrum in Figure L3.1.

3. Is there a relationship between the number of hydrogens attached to a given carbon and the
number of peaks in the cluster associated with that carbon? If so, what is it?
 ChemActivity L3: Carbon (13C) NMR Spectroscopy 285

Memorization Task L3.1: Peak Multiplicity = [Number of H’s] + 1

Chemists say that “the two H atoms attached to Ca (we will call them Ha) ‘split’ the signal of Ca into
three peaks.” This is the multiplicity of a peak and is illustrated in the diagram below for peak clusters
a and b. The dotted line illustrates what the peaks would look like without any splitting.
Hb
Ha Ha
Blow-up
Blow-up
of signal b
of signal a

2 H atoms "split" the signal due to Ca 1 H atom "splits" the signal due to Cb

A peak cluster with…

• one peak is called a singlet (s)
• two peaks is called a doublet (d)
• three peaks is called a triplet (t)
• four peaks is called a quartet (q)
• five or more peaks is called a multiplet (m)

Critical Thinking Questions

4. Label each peak cluster in Figure L3.1 with s, d, t, q, or m.

5. Which of the following best explains the placement of peak clusters along the x axis?
a. The number of hydrogens attached to that carbon atom.
b. The total number of bonds to that carbon atom.
c. Distance from the Br atom (no. of bonds away from Br atom).

Memorization Task L3.2: Memorize the following 13C NMR Chemical Shifts (ppm ranges)
The property measured by the x axis is called Examples ppm
Type of Carbon
chemical shift (think of it as frequency). It is (R = H or alkyl) Range
measured in strange units called ppm. C with all single CH4
5-60
The chemical shift of an atom is related to the bonds to C's or H's R3CH
density of the electron cloud around that atom. C with a single bond R3C—OH
20-90
Chemical shift is hard to predict precisely. to O, N, or a halogen R3C—Br
Memorize these NMR ppm ranges (13C). C with double/triple
R2C=CR2 110-150
bond to C (e.g. C=C)
Factors effecting chemical shift are additive.
E.g. alkene C attached to a halogen (C=C–Br) C with double bond R2C=O
150-220
could be found above 150 ppm. to O (C=O) RCO2H
286 ChemActivity L3: Carbon (13C) NMR Spectroscopy

Critical Thinking Questions

6. Within the category of carbons singly bound to only C and H (5-60 ppm), quartets will appear
between 5-30 ppm; triplets (15-50 ppm); doublets (25-60 ppm); and singlets (30-40 ppm). Are
these refined ranges consistent with the chemical shifts of the peak clusters in Figure L3.1?

13
7. Consider the following C NMR spectrum of 2-bromobutane. Label each peak cluster with a
number (1, 2, 3, or 4) indicating its assignment to a specific carbon and a letter (s, d, t, or q).
Br

1 CH CH3
3 4
2
H3C C
H2

100 ppm 80 60 40 20 0 ppm

13
Figure L3.2a: Cartoon of (coupled) C NMR Spectrum of 2-bromobutane

Model 2: NMR, Coupled versus Decoupled Spectra

If you have a complex molecule with several similar types of carbon atoms, the peak clusters generally
overlap. This makes interpretation very difficult. To solve this problem, chemists "decouple" the H's
from the C's. A decoupled 13C NMR spectrum of 2-bromobutane is shown below.

100 ppm 80 60 40 20 0 ppm

13
Figure L3.2b: Cartoon of Decoupled C NMR Spectrum of 2-bromobutane
Think of each C–H bond as a phone link. In the coupled spectrum, each H is “talking to” the nearest
carbon, “splitting” the carbon signal through this "phone link." To make a decoupled spectrum, we
produce lots of static on the “phone line” so each carbon cannot “hear” the H’s attached to it.
Consequently, each carbon shows up as a singlet (one peak). You will probably never see a coupled
13
C NMR spectrum outside this activity. Their value as a teaching tool will be apparent in the next part.

Critical Thinking Questions

8. What structural information is lost when we decouple the C's from the H's?

9. What basic structural information does a decoupled 13C NMR spectrum still convey?

13
10. T or F: In C NMR the height of a peak is proportional to the number of equivalent carbons
represented by that peak.
 ChemActivity L3: Carbon (13C) NMR Spectroscopy 287

11. Sketch a reasonable (coupled) 13C NMR spectrum for 3-methyl-1-pentene. Then, below it, sketch
the decoupled spectrum. Don’t worry about exact placement or relative order of the peak clusters.
Just make sure each peak cluster is in the right range.

180 160 140 120 100 80 60 40 20 0 ppm

12. The NMR spectra in the previous sections are computer-generated cartoons. Shown below is a
real decoupled 13C NMR spectrum of 2-bromobutane. Label some differences between the
computer-generated decoupled spectrum (Figure L3.2b) and the real version (Figure L3.3, below).

13
Figure L3.3: Decoupled C NMR Spectrum of 2-bromobutane
288 ChemActivity L3: Carbon (13C) NMR Spectroscopy

Memorization Task L3.3: NMR Reference and Solvent Peaks

On a 13C NMR spectrum (also called a CMR spectrum) you will often see a peak at 0 ppm and three
peaks centered at 77.0 ppm. These peaks are associated with additives, not the analyte molecule.
The most common additives are TMS (a reference compound) and CDCl3 (a solvent).

CH3 Cl
TMS (tetramethylsilane) CDCl3 (deuterated chloroform)
a reference compound that is often a solvent that produces three small
H3C Si CH3 Cl C Cl
added to the sample because it is inert peaks centered at 77.0 ppm, where few
and gives a peak at 0 ppm that can be other peaks appear (also can be used
CH3 used calibrate the ppm scale D to calibrate the ppm scale)

Critical Thinking Questions

13. Label the peaks in Figure L3.3 due to TMS and CDCl3 with these terms. Also label the impurity
peak near 50 ppm that is not associated with the solvent, reference, or analyte (2-bromobutane).

Model 3: DEPT NMR

Figure L3.4 is a DEPT NMR spectrum of 2-bromobutane. A DEPT spectrum consist of four spectra:
the top spectrum shows only carbons with three H’s; the second shows only carbons with two H’s; the
third shows only carbons with one H; and the bottom shows all carbons with one or more H.

Carbons with three attached H's (all CH3 carbons)

Carbons with two attached H's

(all CH2 carbons)

Carbons with
one attached H
(all CH carbons)

1 3
2 All carbons with H's
4
reference
peak
Br (TMS)
 ChemActivity L3: Carbon (13C) NMR Spectroscopy 289

Figure L3.4: DEPT NMR Spectrum of 2-bromobutane

Critical Thinking Questions

14. Even though each peak cluster on a DEPT spectrum appears as single peak, you can tell by the
level on which it appears if it is a quartet, triplet, doublet, etc. Label each peak with the letter q, t,
d, or s, and assign the peaks to carbons 1-4 on the structure of 2-bromobutane on the spectrum.
15. Look at the structure of TMS in Memorization Task L3.3 and explain why the TMS peak appears
on the bottom and top levels of the DEPT spectrum.

Memorization Task L3.4: Singlets do not appear on a DEPT Spectrum

By convention, only carbons with H’s appear on a DEPT spectrum. To see all the peaks including the
singlet peaks you must look at an ordinary 13C NMR spectrum.

16. Label each carbon on the structure of 2-butanone with a letter (s, d, t, or q) indicating the type of
peak you expect for that carbon
a. Label each peak on the DEPT spectrum of 2-butanone with a number (1, 2, 3, O

or 4) indicating its assignment to a specific carbon on the structure.

1 2 3 4
b. Which carbon of 2-butanone does not show up on the DEPT spectrum? 2-butanone
c. In what ppm range, and on what type of spectrum do you expect to find the
peak for carbon 2? (Check your work: See Figure L3.5 on the next page.)
290 ChemActivity L3: Carbon (13C) NMR Spectroscopy

Figure L3.5: Ordinary 13C NMR Spectrum of 2-butanone

Model 4: Symmetry Planes

Like all two-dimensional objects, Objects
1-3 have a plane of symmetry (also called a
mirror plane) in the plane of the paper.
For Objects 1-3, some planes of symmetry
are shown with dotted lines. Some are not.
Object 1 Object 2 Object 3

Critical Thinking Questions

17. Mark the planes of symmetry that are NOT marked or noted in Model 4.
a. Object 1 has two additional planes of symmetry. Mark them.
b. Object 2 has one additional plane of symmetry? Mark it.
c. Does Object 3 have any additional planes of symmetry? If so, mark them.
 ChemActivity L3: Carbon (13C) NMR Spectroscopy 291

18. Mark each plane of symmetry in the following molecules.

(For each, indicate if the plane of the paper is a plane of symmetry by writing “POP”.)
OH HO Because of single
Br bond rotation, OH
sweeps out a
C C H H symmetrical CH3
C CH2 O
HC CH HC CH region of space
H2C CH
HC C
Br HC CH HC CH =
C H2C CH
H C C
H H
CH3
Note: Consider the average of both resonance forms and the average
of single bond rotations when considering symmetry.

Model 5: Identifying Equivalent Carbons Using Symmetry

Two carbons are considered NMR (or chemically equivalent) if any of the following are true…
• They are directly across a mirror plane from one another.
• They can be rotated into one another without changing the overall representation of the molecule.
We will add one more item to this list later in the activity.

Critical Thinking Questions

19. Make a model of 2,2,-dimethylpentane, and confirm that BOTH bullets in Model 5 are true for the
three carbons attached to C2. (Note that either bullet would be enough to make these three carbons
equivalent and appear as one peak on a decoupled 13C NMR spectrum).

2,2-dimethylpentane

20. Number the carbons in each molecule in Figure L3.7 to show which ones are equivalent or
distinct. That is, give equivalent carbons the same number. (When you number a molecule to
assign its carbons to an NMR spectrum you do not have to follow IUPAC numbering rules.)

21. For the following molecules, confirm that the plane of the paper (POP) is NOT a symmetry plane.
Cl Cl Cl Br Br Br
Br Cl Br Cl

a. Circle the two molecules that DO NOT contain any symmetry plane. For the others, mark all
planes of symmetry using a dotted line.
b. Number the carbons to indicate which carbons are identical and which are distinct.
292 ChemActivity L3: Carbon (13C) NMR Spectroscopy

Check Your Work: Tips for finding symmetry planes in a molecule

Use the following tips to check your answers to the previous two CTQ’s.
• If there are two of something (e.g., Br groups or methyl groups in Figure L3.7), look for a
plane of symmetry halfway between them.
• If there is only one of something (e.g., OH group—see Figure L3.7), look for a plane of
symmetry that contains that group.

Model 6: Rotational Symmetry Axis

Rotational symmetry axis = an axis around which the molecule as a whole can be
rotated (some distance less than 360o) to result in an identical representation.
Object 3 in Model 4 does NOT have any planes of symmetry (other than the plane of
the paper). HOWEVER, it has a rotational symmetry axis. To test for this axis, put your
pencil on the open dot and spin the paper until the object looks exactly as it did at the Object 3
start. YOUR PENCIL DEFINES THE ROTATIONAL SYMMETRY AXIS.

Critical Thinking Questions

22. Rotating Object 3 _______ degrees (number less than 360) results in an identical representation.

23. Consider the object at right.

a. Mark the rotational symmetry axis with a
b. What is the minimum angle you can rotate this object to
give an identical representation?

24. Carbons that can be rotated into one another via a rotational HO
H
C CH3
symmetry axis are NMR equivalent and chemically equivalent. C C
a. Make this rule a third bullet in Model 5.
C C
b. Does the molecule at right have any plane of symmetry (other H3C C OH
H
than the plane of the paper)? If so, mark it with a dotted line.

c. In this case, does the plane of symmetry in the plane of the paper cause any carbons to be
equivalent to each other? If so, which ones?

d. Does this molecule have a rotational symmetry axis? If so, mark it with a .

e. Number the carbons to indicate which are identical and which are distinct.
 ChemActivity L3: Carbon (13C) NMR Spectroscopy 293

Exercises
1. Often a symmetry plane is coincident with a rotation symmetry axis. Which two dotted lines that
you drew in Figure L3.7 (to represent mirror planes) are also rotation symmetry axes?

2. Draw a representation of 3,3-dimethylhexane so that its symmetry plane is in the plane of the
paper.
a. Circle the two identical carbons on this molecule.
b. True or False: A symmetry plane in the plane of the paper cannot be used to show that two
carbons are identical. If true show this plane for 3,3-dimethylhexane.
3. For each structure:
a. Find each mirror plane and each rotation axis.
b. Number the carbons, giving the same number to equivalent carbons.
c. Label each carbon with a letter to indicate the multiplicity of its 13C NMR peak (s, d, t, or q),
and where it would appear on a DEPT spectrum.

Cl

O
O

O O