A. Protein Precipitation 1. a. protein extract + picric acid b. after addition of 10% NaOH 2. a. protein extract + 0.001 M CuSO4 b.

after addition of EDTA 3. a. protein extract + saturated (NH4)2SO4 b. after addition of distilled water B. Characteristic color reactions of some proteins 1. Biuret Test Protein extract + conc. KOH + 1% CuSO4 GRAY SOLUTION (should be PURPLE SOLUTION) 2. Xanthoproteic test Protein extract + conc. HNO3 + NaOH 3. Ninhydrin test Protein extract + 0.1% ninhydrin BLUE SOLUTION ORANGE MIXTURE WITH PRECIPITATE bright yellow mixture with yellow precipitate orange solution cloudy white with white precipitate cloudy solution cloudy white with white precipitate cloudy solution

C. Separation of amino acids by paper chromatography Chromatographic solvent used: 12:3:5, BUTANOL: ACETIC ACID: WATER Solvent Front Solute Front Computed Rf Amino acid Glu Leu Val Tyr Met Unknown 11.5 cm 11.4 cm 11.3 cm 11.3 cm 11.3 cm 11.3 cm 5.4 cm 6.5 cm 5.5 cm 5.5 cm 0.474 0.575 0.487 0.487 3.5 cm 0.304

D. Solubility and Color reactions of Fats 1. Solubility Fresh coconut oil + benzene Fresh coconut oil + ethanol SOLUBLE; CLEAR YELLOW INSOLUBLE; TOP: ETHANOL BOTTOM: OIL Fresh coconut oil + distilled water INSOLUBLE; TOP: OIL BOTTOM: WATER 2. Characteristic color reaction of Oils and Fats Species Used: Cocos nucifera Endosperm + Sudan III or IV Illustration: (100x) RED ORANGE

Study Questions 1. Why are amino acids considered amphoteric compounds?

2. What is protein denaturation?

3. What is the significance of Rf values? Is it possible to have an Rf value of greater than one and the same Rf s for a single amino acid chromatographed in different solvent? Explain.

4. Name at least three lipid groups in the plant body and give their respective functions.