TCL Flashcards
on what does the separation in TCL depends?
the adsorption to the stationary phase and the partition coefficient
how are the spots visible
the TCL ülat is usually coated with fluorescnet compound
what’S the principle in TCL
different compounds will have different solubilities and equilibrium distribution between the two phases in which tehy are partitioned
on what does the equilibrium distribution depends?
on the nature and degree of interaction of the components with MP and SP
what is silica gel plate used for
for separating weakly to strong polar compounds as it contains OH-groups
two functions of the MP
responsible for the transprt of the sample and secondly, the create separation system
whats the disad of water as MP
it forms hydrogen bonds with the silica plate, thus prolonging the development of the TLC
ad. von water as MP
it has higher elution powder which is needed for the amino acids to elute from the plate
TCL is used as
as a qualitative technique
what is a IEC
exchange column chromatography (IEC) it is a continous flow chromatography where sample containing mixture of amino acids is loaded onto a columne of cation-exchange resin and buffers of different pH and strength pump through the columne to get the separation of AA
when can be used LC in AA
AA need derivatization either pre or pro derivitization
which derivatizing agent can one used?
o-phthalaldehyde, phenly isothiocyante (PITC), fluorescamine and dansyl chorid
what’s the process in the post column derivatization?
AA is first separated in the column and then mix with the derivatizing agent
what’s the benefits of post column derivatization
it offers very good qualitative and quantitative performace and reproducibility
What are some parameter that influenced the outcome of the purity testing by using the TCL
MP, SP, chamber saturation , polarity and also the visual evaluation
