PROCEDURE.

SAMPLE PREPARATION FOR IR ANALYSIS.

A. i) The KBr method

1. Benzoic acid and the KBr solid (IR grade) were obtained from the desiccator. 1 gram
of each were weight and was dried in the oven for about 2-3 hours at 110 °C.
2. The agate mortar and pestle were removed from the desiccator for use to grind the
benzoic acid and KBr.
3. About 1 mg of benzoic acid were took and grinded by using agate mortar and pestle
until the benzoic became very small and shiny.
4. 80 mg of KBr were took and grinded using agate mortar and pestle until it became
powdered.
5. The benzoic acid and powdered KBr were mixed in agate mortar for ration 1:80 and
the mixture were grinded for about 30 seconds until became homogenous.
6. The mixture was scraped into the middle using a spatula and the mixture were grinded
again for about 15 minutes to mix the benzoic acid thoroughly with the KBr.
7. The benzoic acid must be grounded finely, or the mixture will scatter the infrared
radiation excessively.
8. By using a spatula, the mixture was heaped in the centre of the agate mortar.

ii) Making the KBr pellet.

1. The die set were removed from the box or storage container.
2. The die set were cleaned with ethanol.
3. The mixture of benzoic acid and KBr was put into the die set. The mixture needs to
fills the surface of the die set.
4. The die set was tightly closed and was put into Hydraulic Press gauge. The Hydraulic
Press gauge was tightened.
5. The Hydraulic Press gauge was pressed until the pressure goes up to 7000 psi.
6. The air is released and rest for 2 minutes.
7. The Hydraulic gauge again was pressed until the pressure goes up to 8500 psi and rest
for about 1 minutes. The pressure was released.
8. The die set was slowly removed from the Hydraulic Press gauge.
9. The die set was opened and the KBr pellet that was obtained should appear clear like
a piece of glass or transparent disk.
10. The KBr pellet was put into a pellet holder for analysis.

B. Preparing Liquid Sample (neat liquid) using a Salt Plate.

1. A ‘neat liquid’ sample is used, sample need to be dry and no water.
2. Some of the liquid sample (1-2 drops) was put on a plate, and the plate was covered
with another plate. To cover the entire plate the liquid will spread out. Do not press.
To spread the sample the sample in case it does not cover well, try turning the top
plate or add a bit more sample.
3. The sandwich was placed in the IR salt plate holder and was covered with hold- down
plate.
4. At least 2 nuts were put on the posts of the holder and was gently spined to hold the
plates with even pressure. Do not force, it will crack the plates.
5. The holder was slides and placed into a bracket on the instrument in the sample beam.
6. The spectrum was run.
7. The salt plates were removed from the instrument and was cleaned with a little
acetone and dried using Kim Wipes. The salt plates were kept in a clean and dry
place.

C. Sample Preparation for FTIR (Reflectance)

Solid samples must be grounded finely before used.

1. The spectrum of benzoic acid was run using the Reflectance method.
2. An unknown was obtained from instructor and the spectrum was run.
D. OPERATION OF THE FTIR
i) Instrument: Thermo / Nicolet 380 FTIR

Operating Instruction

1. ‘EZOMIC’ program was selected.

2. The icon ‘COLLECT’ was selected and ‘EXPERIMENT SET UP’ was choose, to
analyze a pellet.
3. The data which appeared in the window was checked.
i) No of scan: 32
ii) Resolution: 4
iii) Final format: Transmittance, Absorbance.
iv) File handling: Save interferograms.
v) Background handling: Collect background before each sample.
4. ‘SAVE’ and ‘OK’ was clicked.
5. ‘COLLECT’ was selected and ‘COLLECT SAMPLE’ was clicked.
6. The title of the experiment was type and ‘OK’ was clicked.
7. To collect the background spectrum ‘OK’ was clicked.
8. The pellet was placed into the sample compartment after the scanning is complete.
9. ‘OK’ was clicked to obtain sample spectrum.
10. ‘Find peaks’ and ‘replace’ was clicked after the scan is complete.
11. Went to ‘process’, ‘Advanced ATR correction’ was chose, ‘GE’ then ‘OK’ was
clicked.
12. To identify the compound ‘Analyze’, ’Library setup’,’ Search’ was clicked.
13. The result was saved and printed.
ii) Instrument; Perkin- Elmer Spectrum-RX
Operating Instruction
1. Shift, scanned.
2. The silica gel was removed from the FT-IR.
3. The energy was checked, stop was pressed if the energy >70 %.
4. Scan was pressed again.
5. Background button was pressed. 3 times scanning was chosen.
6. Execute the scan was pressed.
7. Background button was pressed and put on the sample holder.
8. The axis X, Y, or Z was chosen. The screen shown like the background spectrum.
9. Arrows was used to modify the graph.
10. The peak was marked. Shift V cursor or Shift H cursor was used. Then the shift was
marked.
11. Lastly, the plot button was pressed to printout the result.
PRE-LABOTORY QUESTION.

1. Write the molecular formula.

-C9H8O2.
2. Draw the molecular structure.

3. Based on the molecular structure and using Table provided in the introduction (or any
other IR Correlation charts available):
i) predict the location of three peaks you might expect from its IR spectrum.
-1680-1640 cm-1 (m, w)
-3000- 2500 cm-1(b)
-1760-1670 cm-1 (s)

ii) Indicate what bonds were responsible for all the peaks you peak you identified
in (i),
RESULT

1. Observation during preparation of KBr pellet.

The agate mortar that contains KBr was mixed together with the benzoic acid until the
mixture is homogenous. By compressing a hydraulic press gauge the pellet was made.

2. Name of compound analysed.

i) Benzoic acid
ii) Acetone
iii) Caffeine
iv) Plastic
v) Cyclohexane
vi) Unknown

The molecular formula of compound analysed.

i) C7H6O2
ii) C3H6O
iii) C8H10N4O2
iv) (C2H4) n
v) C6H12
vi) C26H42O4
3.
i) Unknown number: vi
ii) Molecular formula of unknown compound: C26H42O4