BIOCHEMISTRY LABORATORY ACTIVITIES

Activity #1
A. Carbon and Hydrogen
-EA—->C+H20
-Heat denature the EA(egg albumin) from high molecular structure to loose,
random ones. EA formed chars when heated does mu black ang surfae
niya, this indicates the presence of carbon and water vapor in the mouth of
test tube indicates the presence of hydrogen.
B. Nitrogen
-EA+CaHNaO2(soda lime)—-->NH3 (ammonia)
-So si soda lime siyay liberator meaning iyang i liberate ang nitrogen
element from the egg albumin with heat, and since soda lime is composed
of an alkaline solution, ma transformed siya into an ammonia which has a
weak base. Si red litmus paper is a confirmatory test of the presence of
ammonia, once turned blue,this indicates the presence of ammonia
C. Sulfur
-
D. Phosphorus
E. Halogens

Activity #2
Activity #3

B. REACTION OF STARCH
1. Reaction with Iodine (IODINE TEST) -a test that determines the presence of
starch. It is also a test that can helpdistinguish starch from monosaccharide,
disaccharide and polysaccharide. This test can only be performed in acidic
conditions as the starch hydrolysis under such circumstances.

Reagent: Iodine in Potassium Iodide in Starch Solution

IODINE POTASSIUM IODIDE STARCH SOLUTION

Color: Yellow Color: Clear Color: Clear

ChemFormula: I2 ChemFormula: KI ChemFormula:

(C6H10O5) n

RESULTS: Brown/Yellow -negative, presence of starch is absent

Blue/Black -positive, presence of starch is present

2. Fehling’s Test -a test used for the detection of reducing and non reducing sugar.
It is also used to differentiate between ketone functional groups and water-
soluble carbohydrates.

Reagent: Sugar solution + Fehlings A & Fehlings B

Fehling’s A -CUSO4, Copper (II) Sulphate

Fehling’s B -KNaC4H4O6.4h2O, Potassium Sodium Tartrate, NaOH, Sodium
Hydroxide

RESULTS: BLUE -aldehyde-ketone is absent

RED -aldehyde-ketone is present

3. HYDROLYSIS OF STARCH
Boiling starch in hydrochloric acid in a diffusion experiment would likely cause the
starch to degrade and break down into simpler sugars. This process is called
hydrolysis. Iodine is often used as a test for the presence of starch, and when it is
added to a solution containing starch, it will typically change color from dark brown to
blue-black. However, in this specific scenario, the iodine would not be able to react with
the starch due to the hydrochloric acid breaking down the starch. Therefore, there
would be no color change observed. It's worth noting that hydrochloric acid is a strong
acid that can cause harm if handled improperly, so it is important to take appropriate
safety precautions when conducting experiments with it.

REAGENT: starch solution + HCl + Heat = Clear (add solution in spot plate and
add iodine)

Concentrated Hydrochloric Acid hydrolyzes or breaks down Starch

IODINE -yellow in color but after some time it becomes clear (achromatic point)
-this means that the starch is now converted into simple sugars or (mono). This
also makes the chemical reaction process change quickly

Neutralized Starch (nabilin sa bath) + Sodium HYdroxide with Fehling’s A and B

BLUE means completely hydrolyzed, NO PRESENCE OF STARCH

Activity #4
 A. Fehling’s Test

Purpose:
- used to identify the presence of aldehydes or groups that contain any aldehyde
functional group -CHO
- also used to differentiate a ketone group and water-soluble carbohydrates.

Reagent: copper sulphate and sodium hydroxide solution

Chemical Formula:
● Copper sulphate - CuSO4 (inorganic compound)
● Sodium Hydroxide - NaOH (often known as Caustic Soda or lye)

Result: Red color (positive, aldehyde-ketone is present)

Blue color (negative, aldehyde-ketone is absent)

B. Benedict's Test
Purpose: This test is used to determine the presence of reducing sugar
Reagent: a complex mixture of sodium citrate, sodium carbonate, and the
pentahydrate of copper(II) sulfate.
Chemical Equation: RCHO + 2 Cu2+ + 5 OH − → RCOO − + Cu 2O + 3 H 2O
Reaction: .

C. Trommer’s Test
Purpose: to identify mainly water-soluble aldehydes and differentiate them from
ketones. The former compounds undergo the reaction, while the latter do not.
Furthermore, the Trommer's test confirms the reducing properties of such
compounds.
Reagent: copper sulfate and potassium hydroxide (base)

ACTIVITY 5

A. BARFOED’S TEST
 Purpose: used to distinguish monosaccharides (simple sugars that have a positive result
with red precipitate) from disaccharides (two sugars that have a negative or slower
reaction).

- It relies on the ability of monosaccharides to react more quickly than

disaccharides with a copper reagent.

Reagent: 0.33 molar solution of copper (II) acetate in 1% acetic acid solution

Reaction: When ang solution nga naga contain og monosaccharide like glucose, galactose, and
fructose if ma heat sha sa barfoeds reagent, ang monosaccharides mo react sa copper ions in
the reagent, mao ni ang gitawag nga redox reaction, where the monosaccharides acts as a
reducing agent and reduces the copper ions in the reagent. The result of this reaction is the
formation of of a red or brick red precipitate meaning positive result sha and nag indicate nga
ang present ang monosaccharides in the tested solution. Disaccharides, which consist of two
sugars do not react as quickly with barfoeds reagent (pwede sha ma positive if ma dugayan og
boil)
2+ +
Chemical equation: RCHO + 2Cu + 2H2O → RCOOH + Cu2O↓ + 4H

B. Picric Acid Test (also known as phenol test)

- an organic compound with the formula (O2N)3C6H2OH. Its IUPAC name is 2,4,6-
trinitrophenol (TNP).

Purpose: a very sensitive chemical test for the presence of reducing sugar. It is another test
for detection of reducing sugars. The reducing sugars react with picric acid to form a red
colored picramic acid.

Reagent: picric acid, or 2,4,6-trinitrophenol, is the critical reagent. Picric acid is a yellow
crystalline compound.

Reaction: wala pa kay galibog pako huhu

C. Nylander’s Test

Purpose: a chemical test used to detect the presence of reducing sugars, specifically
monosaccharides, in a given solution.

- Reducing sugars are sugars that have a free aldehyde or ketone group and can act
as reducing agents in chemical reactions.
- Reducing sugars are sugars that have a free aldehyde or ketone group and can act
as reducing agents in chemical reactions.
- This test is often used in food analysis and other applications where the presence of
simple sugars needs to be identified.

Reagent: Reagent used in the Nylander’s test is the nylander reagent itself. Nylander is a
solution containing a bismuth nitrate (Bi(NO3)3) , potassium sodium tartrate (KNaC4H4O6·4H2O)
and potassium hydroxide (KOH).

Reaction: When a solution containing reducing sugars such as glucose, galactose, fructose,
and maltose, is detected by the formation of a black precipitate or a dark brown to black
coloration.This change in color or the formation of a black precipitate is a positive result, indicating
the presence of reducing sugars in the tested solution. Sucrose lacks a free aldehyde or ketone
group that is required for a sugar to act as a reducing agent. Therefore, when testing sucrose with
the Nylander's test, it should generally yield a negative reaction, with no black precipitate or dark
coloration. Starch does not contain an aldehyde or ketone group. Therefore, when you perform the
Nylander's test on starch, it should typically result in a negative reaction, meaning no black
precipitate or dark coloration will form.

Chemical Equation: Nylander Reagent (containing bismuth subnitrate, Rochelle salt, and sodium
carbonate) + Reducing Sugar → Formation of Black Precipitate or Dark Brown to Black Coloration