6. Chromatography on normal-phase silica gel is based on which of the following
physicochemical processes?
a) Distribution
b) Adsorption
c) Ion exchange
d) Antigen-antibody interaction
Answers
Answer:
I think it would be absorption aur it can be distribution
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Answer:
‘Chromatography’ is an analytical technique commonly used for separating a mixture of chemical substances into its individual components, so that the individual components can be thoroughly analyzed. There are many types of chromatography e.g., liquid chromatography, gas chromatography, ion-exchange chromatography, affinity chromatography, but all of these employ the same basic principles.
Chromatography is a separation technique that every organic chemist and biochemist is familiar with. I, myself, being an organic chemist, have routinely carried out chromatographic separations of a variety of mixture of compounds in the lab. In fact, I was leafing through my research slides and came across a pictorial representation of an actual chromatographic separation that I had carried out in the lab. I guess that picture would be a good starting point for this tutorial!
Let me first explain what I was trying to do here. I had two reactants ‘A’ and ‘B’. I let them react with each other, under certain reaction conditions, to form a product ‘C’. After the reaction was complete, I ended up with a reaction mixture that contained unreacted A, unreacted B and my desired product C. Now my task was to separate out A, B and C to isolate and analyze pure product C.
Illustration of thin layer chromatography (TLC) and glass column chromatography
Illustration of thin layer chromatography (TLC) and glass column chromatography
First, as shown in the left hand side panel, I ran a thin layer chromatography (TLC) plate. This is basically a rectangular piece of glass plate, coated with a thin layer of silica. I applied a spot of the reaction mixture just above the base of the plate (denoted with a solid line), and placed the plate in a jar that contained an appropriate organic solvent (in this case, 1:1 volume by volume mixture of hexane:ethyl acetate was used), with just enough volume to dip the lower edge of the plate. Gradually by capillary action, the solvent started rising up the silica plate, and as you can see the reaction mixture separated into 3 spots with distinct colors by the time the solvent had reached the solvent front mark.
Next, in order to actually perform the separation, I assembled a glass column (as shown on the right hand side of the picture). I took a glass column with a stopcock attached at the bottom, inserted a cotton plug at the bottom of the column and packed the column with a slurry of silica gel (prepared in an organic solvent). Once the column was packed, and the solvent volume above the bed reduced to less than 5 mm, I carefully poured the reaction mixture over the bed of silica from the top of the column, with the aid of a glass pipette. I opened the stopcock and let the solvent run slowly through the column. I constantly kept adding solvent from the top of the glass column. As you can see, the reaction mixture started separating into three distinct bands - yellow, pink and orange corresponding to unreacted B, unreacted A and the desired product C, respectively. I collected individual bands in separate flasks and was thus able to obtain pure C!
Explanation: