I have tried different solvents yet I can observe only 1 spot. The compound seems to be highly polar and I have used different combinations of methanol:ethylacetate:Hexane:acetic acid and butanol still only one spot is there. After this I even used the UV spectrophotometer and I got one not too sharp but one peak. HELP!!!!
You can use different types of Spraying reagent, for highlighting the spot on plate and to detect the presence of bioactive components. Also, try to change your ratio of mobile phase, apply 2-3 different ratio on TLC/HPTLC plates individually. The one is showing the maximum result, you can finalize as your mobile phase.
Are you using SiO2 as stationary phase? The plates were activated (130 deg.C for 1 hour)?
Start with sole n-hexane as an eluent.
Regards
GB
Thankyou Divya Jain for your answer. I have tried and still only one spot is being shown. All the above mentioned solvents have been used in different ratios and still there is no differentiation which is why I am in a fix.
Subecha Rai on which bioactive components class you are focusing for isolation ?
Example- If you want to isolate Flavonoids, then search for flavonoids best Mobile phases for TLC, make a list, prepare small TLC plates for trials, apply all selected mobile phases individually. Which show dark and number of spots, apply on big plate parallely. If they show a number of spots, but not so dark, then try to change their ratio.
Note: Room Temperature, Humidity, and moisture can affect your TLC results, try to perform in cold temp, switch on your lab AC while performing, or you can wait for winter's.
Thankyou so much @Divya I will definitely try to keep all these in mind and go for it.
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