The increase in blue colour is measured in the spectrophotometer at 600 nm.
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TLC
rapid + sensitive method
seperates many compounds of biological + clinical interest
based on differntial partition of solute between a polar liquid phase (help by a thin layer of adsorbet spread on a glass plate) and a less polar moving (mobile) phase.
LIPIDS: the basis of separation is liquid/liquid partition
some separation by a different adsoprtion also occurs as the commonly used silica gel is not completely inert.
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Ninhydrin test
generaly used to detect amino acids and ammonium salts
upon reaction with these compounds a deep blue/purple product is produced (Ruhemanns purple)
Net result = ninhydrin is in a partially reduced form of hydrindantin
The ninhydrin condenses with the ammonia and hydrindantin to produce a purpleish colour
PROLINE:
-reacts with ninhydrin in a different way as the coloour is more yellow
-this is due to a substitution of the alpha amino group that ninhydrin reacts with
-this has to do with the carbon rings.
-Purpleish colour is usually associated with primary amino acids. Where the N is free to react with ninhydrin
-However, in proline the N is not available for reaction as it is locked in the ring structure.
-So no ammonia is produced so now blueish colour is produced.
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