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.