describe the processes involved in the extraction and separation of photosynthetic pigments from a leaf extraction grind leaves in fine sand - to rupture cells and release pigments grind leaves in acetone - extracts large quantity of soluble pigments filter / centrifuge mixture - removes cell debris
separation chromatography - pigments with different solubility carried different distances along paper by petroleum ether solvent repeated spotting of extract - to allow pigment bands to be seen after separation drying between spotting - to prevent spreading of spot paper not touching sides - to ensure solvent rises uniformly spot above solvent level - to prevent extract dissolving in solvent paper left in solvent for suitable time - to allow separation of pigments to occur solvent front marked at finish - solvent evaporates rapidly
calculate a pigment’s Rf value from a chromatogram divide the distance moved from the line of origin by the pigment front by the distance moved by the solvent front