FELIPE, ARIANNE DAPHNE D.

10-12-
22

BSN 1-I

LAB ACTIVITY 9

A. CHEMICAL REAGENTS USED IN EACH TEST

1. BIURET TEST- biuret reagent is made of copper sulphate (CuSO 4), sodium
hydroxide (NaOH) and sodium potassium tartrate (also known as Rochelle salt).
Despite the name, this reagent does not contain biuret ((H 2N-CO-)2NH). It is a
vital component of biuret protein assay.

2. NINHYDRIN TEST- in 1oo ml of ethanol, dissolve 0.35g of ninhydrin solvent for

dilution (for quantitative testing): combine equal parts of water and n-propanol,
standard solution (1 percent protein solution), and sample solution.

3. XANTHOPROTEIC TEST-

- Test solution: 1% tyrosine, 1% tryptophan, 1% phenylalanine, 1% any other

amino acid, protein solution (albumin)
- Nitric acid
- 40% NaOH

4. MILLON’S TEST- millon’s reagent consists of mercuric nitrate and mercurous

nitrate dissolved in nitric acid and distilled water

5. HOPKIN’S COLE TEST

(1) TEST REGEANT- To 10g of magnesium powder add enough water to submerge it.
Then slowly add 250 ml of cold saturated oxalic acid solution. Keep cool. Filter
magnesium oxalate formed. Acidify the filtrate with 25 ml of glacial acetic acid and
dilute it to 1L with water.

- alternatively* glacial acetic acid which has been kept in sunlight for a few minutes
may be used as a test solution.
 (2) TEST SOLUTION- prepare solution containing 2% individual amino acids such as
tryptophan, tyrosine, phenylalanine, glutamic acid, etc. (3) Cone H2SO4

FELIPE, ARIANNE DAPHNE D. 10-12-

22

BSN 1-I

6. LEAD ACETATE TEST-

TEST SOLUTION

-amino acids: 1% solution of amino acids like glycine, cysteine, etc.

- 40% NaOH

-20% lead acetate

FELIPE, ARIANNE DAPHNE D. 10-12-
22

BSN 1-I

B. COLOR DEVELOPED FOR POSITIVE RESULT AND TO WHAT SUBSTANCE IS THE

COLOR DUE
BIURET TEST
-the color turns purple
-all protein and peptides give positive results
-only amino acid, histidine, gives a positive result.

NINHYDRIN TEST

- a positive result shows by the appearance of a complex with a purple color in the
test tube. This coloration denotes the presence of an amino acid in the sample.

XANTHOPROTEIC TEST-

- color changes to orange indicating the presence of aromatic amino acids.

MILLON’S TEST

a positive result in the Millon’s test is demonstrated by the formation of a red or

pink colored precipitate. This indicates the presence of tyrosine or tyrosine
containing protein.

HOPKIN’S COLE TEST

A purple ring appears between the two layers due to presence of tryptophan.

LEAD ACETATE TEST

- formation of black precipitate indicate the presence of sulfur-containing amino

acid.