Judahnyah Clare

Datasheet 8

Results:
Biuret Test

Substance Observation

Egg albumen The solution turns light purple (positive test)

Casein Turned bright purple (positive test)

Amino acid Turned clear blue (negative test)

A biuret test is used to detect proteins because it can detect peptide bonds between amino acids,
the structural unit of proteins. This works because The mechanism copper (II) binds to protein
peptide nitrogen atoms. This assay was used for protein determination in entire tissue samples
without being affected by amino acids. Ammonium sulfate ((NH4)2SO4) precipitation purifies
proteins, but buffers like ammonia interfere with biuret assay. Copper (II) ions react with peptide
nitrogens to displace peptide hydrogens (as long as the environment is sufficiently alkaline).
A chelate complex is formed when four nitrogen atoms donate lone pairs to create coordinate
covalent connections with the cupric ion. 540nm light absorbs this chelate complex, turning it
purple. Proteins in the analyte generate a purple complex. The purple color comes from the
analyte’s peptide bond concentration. A positive result occurs if the sample tested color changes
to purple. Conversely, if the sample solution remains light blue, the protein concentration is low
and considered a negative result. In the experiment, only two substances tested positive (egg
albumen and casein), while the amino acid tested negative because the amino acids were free
without peptide bonds. Biuret molecules are similar to peptides because they are both formed by
two concentrations of a substance.
Ninhydrin test

Substance Observation

Egg albumen The solution turns purple (positive)

Casein The solution turns a violet color (positive)

Amino acid Turns light yellow (negative)

Analysis II:
Ninhydrin is a chemical that reacts with amino acids in proteins to produce a purple color. The
ninhydrin test detects amino acid content in unknown compounds. It is also used in solid-phase
peptide synthesis to ensure that proteins are adequately stored for amino acid analysis. Because
of its high sensitivity, the ninhydrin test is frequently used to detect fingerprints.

Fig1 Ninhydrin structure

Fig2 Nynhydrin and amino acids in proteins reaction
Fig 3 Ninhydrin and amino acid reaction
Fig 4 Ninhydrin and ammonia reaction
III. Heavy metal salt precipitation

Substance Observation

Silver nitrate Heavy white participate forms

Mercuric chloride The solution turns milk white

Analysis III:
1. Describe how silver and lead ions react with proteins. How is this reaction related to these
metals' toxicities?

-The silver and lead ions react with proteins by separating the protein from the metal ions'
solution. Protein dissociates into protein anion (Pr-) on the alkaline side of the isoelectric pH
scale, which interacts with positive metal ions (cations) to generate insoluble metal proteinate
precipitates like lead albuminate and silver albuminate. These heavy metals' reactions are
transported and segregated into frame cells and tissues in the human body. They attach to
proteins and nucleic acids, which break down the macromolecules and impair cell function.
Heavy metal poisoning may therefore have various impacts on the human body.

2. Treatment for ingestion of silver or lead is egg white or milk consumption. Explain why

-Milk and egg white are an antidote for heavy metal poisoning because they consist of proteins
with sulfhydryl (-SH) groups in their structures, which can attach to heavy metal ions and help
the body get rid of them. Heavy metal is removed from the body by vomiting after being
absorbed by egg whites and milk, which also balances stomach acid.