Biochemical Techniques and Instrumentation

Materials

1. Whatman No.1 filter paper
2. Chromatography chamber
3. Hair – dryer or spot- lamp
4. Sprayer or Atomiser
5. Microsyringe or micropipette
6. Mobile Phase (Solvent System): Mix n-butanol, glacial acetic acid and water at the ratio of 4 : 1 : 5 in a separating funnel and allow to equilibrate for 30min. Drain off the lower aqueous phase into a beaker. Place the upper organic phase inside to saturate the chromatography chamber and use it for developing the chromatogram.
7. Standard amino acids : Dissolve different amino acids in distilled water separately at a concentration of 1mg/mL. To dissolve the free amino acids tyrosine and phenylalanine use very dilute HCl (0.05N) solution. To dissolve tryptophan, use very dilute NaOH (0.05N) solution.
8. Ninhydrin Reagent : Dissolve 100mg ninhydrin in 100mL acetone.
9. Elution Mixture : Prepare 1% copper sulfate solution. Mix ethanol and copper sulfate solution in the ratio 80:20(v/v).