Experiment No: 1

Experiment Name: Analysis of samples using Fourier Transform Infrared

spectroscopy.

I. AIM:
Detection of functional groups of (sample name ) using FTIR.

II. Procedure :
A. Steps for sample preparation:
1. KBr pellet method
2. Put KBr powder in oven at 105° C for 1 hr to remove moisture.
3. Clean the parts of pellet casting die, spatula, mortar and pestle with acetone and tissue paper to remove
any type impurity.
4. Mix KBr powder and sample powder in the ratio of 100:1.
5. Grind the above mixture for 3-5 min to get a homogenous mixture.
6. Now with the help of spatula put the grinded mixture in the casting die and press with piston rod
provided.
7. Place this casting die in hydraulic press and maintain the pressure upto 1.5 ton. Wait for 1 min and
remove it.
8. Now gently open the casting die to get the get a thin film of KBr mixed sample.
9. Put this KBr film in the pellet holder and place the holder in the instrument.
10. Now follow the step 11 to 14 from the next section to get the spectra.

B. Steps for using software (if any):

1. Switch on the instrument and PC.
2. Click on IR solution
3. Click on measure tab.
4. Click for initialization, and then click on yes.
5. Check three green signal boxes of humidity, temperature and laser.
6. Go to measurement mode, click on transmittance mode and set no. of scans: 45(>20) and Resolution 4.0
7. Select analysis range: 340 to 7800 cm-1.
8. Click on BCK for Background correction: put KBr pellet in the instrument before clicking (prepare KBr
pellet without addition of any sample).
C. For sample scan
1. Click on Measurement: write sample name in comment.
2. Data file: chose the location to save the file
3. Put sample (KBr mixed) in instrument.
4. Click on sample to get the spectra.
5. To get the spectra data, click on file tab then click on export and save file as ascii file. Direct print of
the spectra can be taken from the print option.

III. Result and Analysis

1. Instrument name (Make and Model):
2. Sample name :
3. Sample amount
4. Sample type(Solid/Liquid/Gas)
Plot Spectrum :

Table 1: Observations

Peaks (cm-1) Attributed to

Discussion:

IV. Precautions to be taken :

1. Never turn off the main power of the instrument as power is required to keep the instrument free from
moisture or humidity.
2. Always use oven dried KBr for sample preparation of IR grade.
3. Always clean the KBr die, spatula, mortar and pestle with acetone before preparation of sample.
4. Never spill the water or liquid on the instrument.
5. Never used pen drive to collect data, always use blank CD.

V. Example analysis
Aim : Detection of functional groups of Spirulina biomass using FTIR

Result and Analysis :

1. Sample name : Spirulina biomass
2. Sample amount: less that a pinch

Spectrum :

90
C-Br or C-Cl
85

80
670 OH
75
% Transmitance

1412
70 2920
1048
C-O
65
C=C C-H
60 1639

55

50
OH and N-H
45 3434

500 1000 1500 2000 2500 3000 3500 4000

-1
Wave number, cm
Table 1: Observations

Peaks Attributed to
3434 OH or NH
2920 C-H
1639 C=C
1412 OH
1048 C-O
670 C-Br or C-Cl

Spirulina biomass is organic material and functional groups present are shown in the above
table.