Thin Layer Chromatography


In this laboratory brain extract, cheese, milk, mayonnaise were used to identify their lipid components. 

Materials and Methods: 

Solution A(chloroform/methanol 1:1) was added as 500µl on each sample and vortexed. Eppendorfs were centrifuged for 3 minutes. Supernatants were transferred to falcons. 500µl solution B  (chloroform/methanol 3:1) was added on pellets and vortexed. Then centrifuged for 3 minutes. Supernatants were collected on their own falcon. Solution C( chloroform/methanol/1,2 M HCl 10:10:1) was taken as 500 µl and added to pellets and centrifuged for 3 minutes. Supernatant were fresh falcon and centrifuged for 3 minutes. . Then samples and standards were put on silica membrane drop by drop. After TLC, membrane washed with 30% sulfuric acid and heated. 

Results: 

 

Samples  a value- Distance (cm)  Rf value a/b b=7,7 
Diacylserine glycerol  5  0,649351 
Phosphotidyl ethanolamine  4,6  0,597403 
Glyceryl trioleate  7,5  0,974026 
Cholesterol     
Palmitic acid   7,5  0,974026 
Brain extract  6,7  0,87013 
Milk     
Cheese  4,4  0,571429 
mayonnaise  7,5  0,974026 

 Discussion: 

Interaction between component an polar silica gel can affect movement of the component. Strong polar interaction between component and silica gel cause less movement toward upward. 

Highest Rf value is belong to most hydrophobic, so glyceryl triolate palmitic acid and mayonnaise are most hydrophobic.Cheese is lest hydrophobic than others so it may contains glycryl trioleate and palmitic acid. 

References: 

  1. http://chemistry.syr.edu/totah/che276/support/6a1.handouts/4.DataInterp.pdf 
  1. http://www.chem.ucla.edu/~bacher/General/30BL/tips/TLC1.html 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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