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Author Topic: QUALITATIVE ANALYSIS OF RAFFINOSE AND SUCROSE CONTENT OF SOYBEANS WITH TLC  (Read 5111 times)
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Francis Umeoguaju
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« on: April 10, 2010, 09:13:44 am »




REAGENTS
   Soybeans
   91 and 71% Isopropyl alcohol
   TLC plates (pre-coated commercial plates)

PROCEDURE

Extraction of sample: 5 grams of powdered soybeans was blended in 10ml of water. The resultant slurry was filtered through the muslin cloth. Equal volume of 71% Isopropyl alcohol was mixed with the extract. This was centrifuge and the supernatant was concentrated by boiling over the Bunsen burner.

The concentrated extract was then centrifuged to obtain a clear extract.

Standard solutions of sucrose and raffinose was prepared by dissolving 2mg of each in 50ml of water.
   
The sample and standards were spotted on the TLC place and was ran with a mobile phase of 91%  isopropyl alcohol.
   
The color was developed by spraying with the carbohydrate spray reagent suggested by Murray (1973).  The sprayed plate was heated over the oven to visualize the bands.
   
The resultant chromatogram was scanned and analyzed with the UNSCAN IT software.

RESULT
   
The result obtained indicates the presence of sucrose and Raffinose. One more band was not identified. This would probably be stachyeous.
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