I have peak area of pure rhein compound and on same rf value peak area of crude extracts. Does anyone know the formula?
If you have pure reference standard of rhein and you have developed HPLC method. The peak in the crude extract of rhein at same rf value should be confirmed first wetehr it is of rhein or something else.
First compare the absorbance value of peak of pure rhein with that of peak in crude extrcat.
If both peaks have same absorbance max. that add +ve control in the crude extract and then see the increase in peak area.
If it works than u can start calculating the amt of rhein in sample.
For this make a calibration curve of rhein with peak area at differnt concentrations and then calculate the amt of rhein from sample by putting peak area in regression equation. In this way you calulate the amount in particular concetration.
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