Hi Revathi

Usually phenol/chlorofom combination is used as a phase separation technique in extracts. Aqueous samples can be mixed with equal volumes of a phenol/chloroform (similar dielectric constants and chloroform is not polar protic like water) mixture. Subsequent to mixing the mixture is centrifuged and two distinct phases are formed because the phenol/chloroform mixture is immiscible with water. The aqueous phase is on top because it is less dense than the organic phase of phenol/chloroform.

Total phenols were quantified better by adding known volume ofstandard NaOH solution then titrate the remind NaOH with standard Soln. of HCl