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
What are other methods which can be used in place of just the TCL
TCL scanner, which is a quantitative analysis or HPTLC
For what is the HPTLC important?
it can be used for QC of larger sample numbers in a short time
What’s the disadvantage of the GC
the sample has to be vaporizatied
- What’s the advantages of a HPLC compared to a TLC?
it is the degree of automation and the low detection limit
quantification and qualification of the substance are possible
- What’s the advantages of a HPLC compared to a TLC?
increase the efficiency of the analyticl method and decrese deviation which leads to a higher reproducibility
- What’s the advantages of a HPLC compared to a TLC?
TLC( plate) is nout reusable
what’s the disad. of HPLC
it needs a higher effort for method development
Different in the separating method of TLC and IEC
AA are separated according to their charge while in the TLC they are separated according to their diference in polarity
