CHM2211L:

EXPERIMENT 1

Experiment 1:
Identification of unknown alcohol based on 1H and 13C NMR
analysis

Purpose
This exercise illustrates how 1H and 13C NMR spectroscopy is used to elucidate the structure of organic
compounds. Students will gain hands-on experience with running benchtop 42.5 MHz Spinsolve Carbon
NMR spectrometer (Magritek). The obtained 1H, 13C NMR spectra supplemented with provided chemical
formula and DEPT-135 data experiments will give students chance to elucidate structure of unknown
alcohol.

Pre-lab preparation
1. For the basic introduction to NMR spectroscopy please read chapter 5.3.3 (pages 159-168)
from Experimental Organic Chemistry Laboratory manual available at:
http://site.ebrary.com/lib/floridaatlantic/detail.action?docID=11117718
This chapter will give you introduction to 1H and 13C NMR spectroscopy.

2. Watch additional introductory videos (clickable links)
a) Mandatory to watch video: NMR Spectra acquisition using Spinsolve. This video is available
on Blackboard under CONTENT > VIDEO TUTORIALS folder. You have to watch it number of
times until you are comfortable with performing the data acquisition. Detailed instructions
are provided below. A mandatory quiz needs to be taken before your lab session.
b)

Figure 1. Screenshot of NMR spectra acquisition video

c) VIDEO: Spinsolve Ease-of-use (https://vimeo.com/128825562)
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3. In your notebook draw a flowchart showing all procedures required for this experiment.
Remember to include spectrometer preparation step.

In the laboratory

At the beginning of the lab your TA will talk about analysis of NMR spectra and structure elucidation.
Please pay close attention and take notes as this information will be necessary for you to complete this
exercise. Next, each student will run their unknown alcohol sample. For this experiment we will be
using neat alcohol sample it means that no deuterated solvent will be present. Your TA will set up
and insert the sample for you. Next, you will run Spinsolve software to obtain:
a) 1H NMR spectrum
b) 13C NMR spectrum.
Once you finish your runs you will be given a pre-run DEPT-135 spectrum of your unknown alcohol.

NMR STRUCTURE
DETERMINATION

Try solve the Analyze 1H NMR

structure signals

Take into account Analyze 13C NMR

chemical formula signals

Analyze DEPT-135
signals

Figure 2 General method for NMR based structure elucidation used in this experiment.

While working on structure determination, in your laboratory notebook write down procedures and
observations and create the NMR signals tables (separate for 1H, 13C and DEPT-135 spectra. Try to
draw your hypothetical structure and see if it make sense (based on the data that you have)

For 1H NMR spectrum:
Atom Atom is Peak Peak For DEPT-135 spectrum:
part of a Multiplicity observed Atom Atom is Peak Peak
group (ppm) part of a observed phase
A CH2 doublet 1.23 group (ppm) (+/-)
1 CH 2 40.2 +

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For C NMR spectrum:
Atom Atom is Peak
part of a observed
group (ppm)
1 CH2 40.2


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Spinsolve NMR:
1
H and 13C NMR spectra acquisition instructions

Introduction
Before your arrival the Magritek NMR spectrometer will be prepared for you. Initially, the
magnetic field inside an NMR spectrometer will be far from homogeneous compared with an "ideal"
field of the device. This is a result of production tolerances and of the magnetic field of the environment.
Iron constructions in walls and floor of the examination room become magnetized and disturb the field
of the scanner. The probe and the sample become slightly magnetized when brought into the strong
magnetic field and create additional inhomogeneous fields. The process of correcting for these
inhomogeneities is called shimming the magnet, shimming the probe or shimming the sample,
depending on the assumed source of the remaining inhomogeneity. This will be done for you.

Please watch tutorial video on Blackboard to familiarize yourself with spectra
acquisition process.

Spinsolve instructions

Your TA will insert the NMR tube into NMR spectrometer for you. Once inserted you may obtain your
desired set of spectra.

A. Obtaining 1H NMR spectra

In the SPINSOLVE software:
4. Click 1H NMR mode.

5. Click protocol. Select following parameters:

6. Solvent selection must be set to NONE.
7. Provide your sample name
8. Select QUICKSCAN
9. Click button to acquire your spectrum. It will take only 10 seconds!
10. After acquisition is complete you will transfer your spectrum to processing software called

MNOVA. Please click on icon to do it.

In the MNOVA software:
11. Your transferred 1H NMR spectrum will be automatically processed and partial processing will
be done for you using built-in automatic functions. Your peaks will be labeled, integrated and
summarized for you.
12. Please print out your spectrum using CTRL + P keyboard shortcut or press printer icon.

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B. Obtaining 13C NMR spectra

In the SPINSOLVE software:

1. Go to the main MENU. Click .

2. Click 13C NMR mode.

3. Click protocol. Select following parameters:

4. Solvent selection must be set to NONE.
5. Provide your sample name. It will be copied automatically from last (1H in this case)
experiment.
6. Select number of SCANS to be 64.
7. The repetition time should be set to 2.
8. Click button to acquire your spectrum. Please wait as 13C NMR spectra
take much more time to acquire! You may start thinking of your 1H NMR spectrum assignment
already!
9. After acquisition is complete you will transfer your spectrum to processing software called

MNOVA. Please click on icon to do it.

In the MNOVA software:
10. Your transferred 13C NMR spectrum will be automatically processed and partial processing will
be done for you using built-in automatic functions. Your peaks will be labeled automatically.
Disregard automatically generated integrals. You have to perform manual peak integration.
11. To manually integrate peaks, press I key on keyboard. The cursor will change from arrow to
integral symbol . Now you will have to click left mouse button and drag it to select integration
area around the peak. The integral value will be given below the ppm axis. Note: the first peak
you choose to integrate will have a value of 1 proton. This can be changed, if necessary.
12. If your peaks are not labelled/integrated correctly, please ask you TA for help.
13. Please print out your spectrum using CTRL + P keyboard shortcut or press printer icon.

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NMR spectral analysis

Please trim your printed spectrum so that it can be inserted (glued/taped) into your notebook.
Lets look at the spectra and try to analyze them.1 Lets look at 1H NMR spectrum first.
Can you identify any obvious impurities or solvents that might be present? (Do we have a solvent
in this experiment?) Crossing them off at this stage is a valuable exercise in data reduction and
clears the way ahead so that you can concentrate on the important peaks.
Can you identify a signal that gives a clear integration for a known number of protons? Now work
from left to right, assigning each signal, or groups of signals that you observe, to protons in your
proposed structure. (If there is logic in starting at the left of the spectrum, it is that most
molecules have some aromatic or heterocyclic core, to which, various alkyl functions are
attached. If there is a problem with the core, then you will at least discover it promptly and be
able to relate it to the alkyl components of the molecule.)
Cross-examine each and every signal in your spectrum to check that they conform to the
expected values for the three crucial NMR parameters: (1) chemical shift, (2) coupling pattern
and (3) integration.

Figure 3. Sample 1H NMR spectrum. Please note peak labels and integrals.

13C spectra analysis.
Carbon NMR spectrum will give you information on carbon chemical environments present. These are
decoupled spectra so no splitting information is present. Quaternary carbon's frequently give small
peaks. Acquisition conditions are not optimized for integration.

1
Richards, S. A., and Hollerton, J. C.. Essential Practical NMR for Organic Chemistry (1). Hoboken, GB: Wiley, 2010. ProQuest
ebrary. Web. 24 August 2016.
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A DEPT-135 spectrum will help you distinguish CH3, CH, CH2 carbons. DEPT-135 stands for Distortionless
Enhancement of Polarization Transfer using a 135 degree decoupler pulse. This pulse sequence produces
a carbon spectrum with methyl (CH3) and methyne (CH) carbons are up. Methene (CH2) carbons are
down. Tertiary carbon atoms are not visible in this kind of spectrum.

Figure 4. Sample 13C NMR spectrum (top) and DEPT-135 (bottom) one.

Chemical formula the missing piece of information.

You are given a chemical formula of your unknown alcohol sample. Together with integrated 1H NMR
signals and 13C analysis you will be able to deduce the structure of your alcohol. Its like chemical Sudoku!

Here are the chemical formulas of your unknown samples:

Sample Chemical
number Formula
1 C3H8O
2 C3H8O
3 C4H10O
4 C4H10O
5 C4H10O
6 C5H12O

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Laboratory report

NMR data analysis can be a difficult task. Chapter 5 from book1 available at:
http://site.ebrary.com/lib/floridaatlantic/detail.action?docID=10441485 will help you to analyze your
NMR data.
Based on 3 NMR spectra (1H, 13C and DEPT-135) and provided chemical formula you will:
1. Elucidate the structure of your unknown alcohol.
2. Assign all proton signals (where possible) to a chemical structure. Please use letters AZ going
from left to right on the structure drawn on the spectrum.
3. Assign all carbon signals to a chemical structure. Please use numbers 1n going from left to
right on the structure drawn on the spectrum.
4. Provide IUPAC or common name of identified alcohol.
5. Additionaly, at home: predict 1H and 13C NMR spectra using on-line tools and compare them
with your experiment.

The website for 1H NMR prediction is here :
http://www.nmrdb.org/new_predictor/index.shtml?v=v2.53.5
The website for 13C NMR prediction is found here: http://www.nmrdb.org/13c/index.shtml?v=v2.53.5

The use of those website is pretty straightforward: Just draw your molecule and click Calculate
spectrum. Please see example below:
Step 1: Draw chemical structure of your molecule
Step 2: Click Calculate spectrum. Your spectrum will be generated.
You can interact with the spectrum (zoom, scroll) by moving cursor over peaks or particular atoms in
the formula window. Your corresponding peaks will be highlighted.

Figure 5. Screenshot from the nmrdb.org website.

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What is expected on the report

Format: 1-inch margins, Font size max 12, Line spacing 1.15 throughout the text.
1. Your laboratory report must contain the following sections (do not simply answer the following
questions these are just hints!):
Title of the experiment
Title clearly identifies the research question
Abstract
What have you done? What methods were used? What have you concluded from your data?
Introduction
Provides background information on the theoretical problem how to solve structures of organic
molecules? 250 words max.
Results and discussion
This section is to be written in first person (plural) form (e.g. we, ours).
Results:
drawn structure of your alcohol (must be drawn on the spectrum)
labels of all proton and carbon signals AZ, 1n
tabular comparison of your observed and calculated spectrum peaks (for both 1H and 13C)


For 1H NMR spectrum:
Atom Atom is Peak Peak Peak
part of a Multiplicity observed calculated
group (ppm) (ppm)
A CH2 doublet 1.23


For 13C NMR spectrum: For DEPT-135 spectrum:
Atom Atom is Peak Peak Atom Atom is Peak Peak
part of a observed calculated part of a observed phase
group (ppm) (ppm) group (ppm) (+/-)
1 CH2 40.2 1 CH2 40.2 +



Discussion:
Discussion of your signals (proton, carbon) and relationship between observed and calculated
peaks and provided chemical formula (similarities/differences, overlapsetc). Do not simply
mentioned them discuss your reasoning in structure elucidation. Note that predicted
spectrum may slightly vary. Why? Especially the OH peak signal may not be seen/predicted
correctly. Why? Add these points to your discussion as well. Remember: the discussion section
always compares ones findings with existing literature!

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Conclusion(s)
In this section you summarize the main features of the report, the objectives, findings and conclusions.
You briefly explain any possible sources of error relevant to your results. Have you successfully
determined the structure? Why/why not?
References
Please cite any references in ACS format. Remember to cite every source that helped you/you reference
to.

2. Attach original NMR spectra and original pages from your lab notebook to a hard copy report.