Add 1 mL dH2O

Add 1 mL saturated NaHCO3

Add 0.25 mL ethanolic sol’n DNFB (0.05 mL DNFB

per mL ethanol)

Shake the mixture

Adjust pH to about 8 using solid NaHCO3
Place a marble on top of the test tube
Heat sample for 15 minutes in a sand bath at
 Cool and subject to liquid-liquid
extraction

Add 2 mL of ether

Shake

Remove the ether layer

Repeat the extraction with

aqueous layer twice

Adjust to pH 1 with 6N HCl

Add 1 mL of saturated NaHCO3

Add 0.25 mL DNFB reagent

Shake the mixture

Adjust pH to about 8 using solid NaHCO3

Place marbles on top of the each test tube

Place marbles on top of the each test tube

Cool samples
Extract with 2 mL portions of peroxide free e
until yellow color is removed
Remove the ether carefully with a pipette or
94 mL Dichloromethane 5 mL Methanol 1 mL Acetic Acid

Mix well and pour in a filtered paper-lined chromatography

tank

Stand for 30-60 minutes to equilibrate

Spot about 0.05 mL of the sample and standards to

previously-prepared thin layer plates

Place in the tank

Bright yellow DNP-amino acids