COLLEGE OF ARTS, SCIENCES and TECHNOLOGY

Name: Mary Angelique Caligan Date: ________________

Section: VETM1C Group #: 5
Members:
Castillo, Alliah
Enriquez, Aloedia
Monreal, Charmae
Ramos, Trinity

Instructions:
1. When answering questions, do not repeat or copy from the laboratory sheet. You are encouraged to
use the laboratory sheet as a reference, but plagiarized answers will not be graded.
2. Do not answer the prelab with a partner – answer using your own words.
3. Do not remove instructions or any bolded text in this prelab sheet, but you are free to edit the bullets
provided.
4. Prelab sheet will be submitted in the assignment tab provided for this experiment .

BIOCHEMISTRY LABORATORY
BCHE102B
PRE-LABORATORY ACTIVITIES

Experiment #4 Analysis of Lipids

(Experiment Number and Title)

I. Safety Measures

1. Put on proper personal protective equipment like goggles, gloves, lab coat, and
mask.
2. Make sure the materials and equipment are clean and in proper condition.
3. Remember the proper handling of materials. Handle the hot test tubes with a test
tube holder and beaker tongs for hot beaker.
4. Before each test ensure that the medicine droppers are rinsed very thoroughly.
5. Follow the procedure diligently and ensure the right amount of samples and
reagents.
 COLLEGE OF ARTS, SCIENCES and TECHNOLOGY
6.
Use gloves when dispensing reagents to avoid burns from the acid or base, especially
when handling the nitric acid.
7. Do not inhale the vapors.
8. Wear gloves and avoid spillage when dispensing the molybdate reagent since it
contains sulfuric acid.
9. Clean the area and check if the materials are placed orderly and the solutions are
stored properly.
II. Materials and Equipment
Lab Materials  Samples  Reagents 

 8 pcs test tube   Corn oil   6M Nitric Acid (80mL) 

 1 pc test tube rack   Butter  6M NaOH (80mL) 
 2 pcs test tube holder   Egg Yolk  Molybdate solution
 Hot plate   Margarine (80mL) 
 Cheesecloth  Coffee Creamer  5 pcs Vitamin C tablet
 Dropper  Olive oil or any for Ascorbic acid
 Mortar and Pestle  vegetable oil solution 
 1pc 250 mL Beaker   30g Potassium hydrogen
sulfate (solid) 
 Distilled Water 
 

III.Procedure. Provide a bulleted list of tasks for this experiment.

1. First, prepare the necessary materials and equipment afterwards, make sure
that the work area is clean and appropriate for the experimentation.
2. There will be two tests, the first procedure is the Phosphate Test. Prepare the
six clean and dry test tubes with labeled 1 to 6 then add about 0.2 g of sample
to each. Add 3 mL of 6 M nitric acid to each test tube to hydrolyze the
compounds.
 COLLEGE OF ARTS, SCIENCES and TECHNOLOGY

3. Prepare the water bath in a 250-mL beaker on a hot plate by boiling an

estimated 100mL of tap water in it. Next is to place the 6 test tubes inside for
5 minutes. After cooling, add 3 mL of 6 M NaOH to neutralize the acid then
mix.
4. A precipitate may form during hydrolysis, particularly in the egg yolk sample.
The precipitate-containing samples must be filtered. On top of a 25-mL
Erlenmeyer flask, place a piece of cheese cloth. Pour the turbid hydrolysate
on the cheese cloth and filter the liquid through it.
5. Fill a clean and labeled test tube with 2 mL of each neutralized (and filtered)
sample. Fill each test tube with 3 mL of a molybdate solution and stir well.
Place the test tubes in a boiling water bath for 5 minutes and allow them to
cool to room temperature.
6. Afterwards, add 0.5 mL of an ascorbic acid solution to each test tube and
thoroughly mix the contents. Wait 20 minutes for the purple color to emerge.
A purple hue indicates that the sample is a phospholipid, while a negative test
indicates fat and cholesterol. Record your observations.

7. The next test is the Sudan IV Test. Repeat the first procedure as the
Phosphate Test. In a six, clean and dry test tubes put 0.2g of each sample to
each test tubes. Add each tube with 4-6 drops of Sudan IV. Place your thumb
over the top of each tube and shake it several times to mix the tube contents
with the water. After mixing, check the tubes again after 1 minute, 5 minutes,
and 30 minutes. Look for a red-orange layer near the meniscus (the liquid's
convex upper surface).
8. Sudan IV will stain the surface layer where the fats are floating or the entire
sample if the fats are mixed into the liquid (emulsion) if lipids are present. A
positive test is indicated by the presence of red-orange color in the solution.
9. Record all your observations.
10. Clean the area and materials after use. Ensure proper storage to each samples
and reagents.
 COLLEGE OF ARTS, SCIENCES and TECHNOLOGY
IV. Mater
ials Safety Data Sheet (MSDS). Search for the MSDS of every chemical included in this
experiment. Report the health hazard ratings, potential hazard, and prevention method
for the chemicals.
 Nitric Acid

- Health hazard ratings:

- Potential Hazard: It can cause severe eye and skin irritation. It can also damage the
respiratory and gastrointestinal tracts.
- Prevention method: Wear proper PPE, goggles to avoid damage to the eye, lab gown
and gloves to avoid skin irritation. Ensure the proper disposal and close container
when not in use.
 NaOH
- Health Hazard Ratings:
- Potential Hazard: Corrosive to metals, it can cause skin irritation, serious eye
damage, eye irritation and irritation to respiratory.
- Prevention Methods: Avoid ingestion and breathing dust/fume/gas. Avoid contact
with eyes, skin or clothing. Wash face, hands, and any exposed skin after handling. Proper
container and make sure the area is well-ventilated and clean. Keep away from
incompatible materials and oxidizing agents.
 Molybdate Solution
- Health hazard ratings:
- Potential Hazard: May cause skin and eye irritation. It can be harmful if inhaled and
ingested. It may cause dermatitis. It may irritate the nose and throat.
- Prevention method: Avoid spillage and store in an appropriate container. Make sure
that the area is well-ventilated. Wear masks, lab gown, goggles and gloves for protection.
Avoid inhaling vapors and wash before and after use.
 Potassium Hydrogen
- Health hazard ratings: Health (3), Instability (2)
 COLLEGE OF ARTS, SCIENCES and TECHNOLOGY
- Potential Hazard: It
respiratory tract.
- Prevention method: Wear nitrile gloves, use safety glasses, masks and lab gown.
Tightly close the container and store in a dry place. Avoid draining to the sink and avoid
spills.

Do not inhale. Clean area before and after use. Avoid contact to combustible or organic
materials.

PRE-LAB QUESTIONS 
1. Lipids are defined by property rather than structure. What is the ‘‘defining
property’’? 
- Lipids are organic compounds composed of fats and oils. They are insoluble
in water and soluble in non-polar solvents examples are ether and
chloroform.
 
2. What are the three groups of lipids and how do they differ? 
- The 3 groups of lipids are triglycerides or fatty acids, phospholipids, and
steroids. Triglycerides is the most common, they are fats and waxes that are
esters of fatty acids with different alcohols. Complex lipids are esters of fatty
acids and contain an alcohol and one or more fatty acids. Examples are
phospholipids and glycolipids. The last one is steroids which have 4 carbon
chains with variety of different side chains. They are substances that
obtained through hydrolysis of lipids.
 
3. Below are structures of several steroid hormones derived from cholesterol. Circle
the steroid nucleus in each. 
 
 COLLEGE OF ARTS, SCIENCES and TECHNOLOGY

 
 
4. Would any of the above give a positive Lieberman-Burchard test? Explain your
answer. 
- Yes, they would give a positive Lieberman-Burchard test because all of them
are steroids which also contains cholesterol. Lieberman-Burchard test uses
acetic anhydride and sulfuric acid to detect the presence of cholesterol by the
appearance of green color as its positive result.
 
5. Cholesterol is an alcohol. Alcohols dehydrate by losing the elements of HOH and
forming a carbon-to-carbon double bond. Draw the structure cholesterol forms upon
dehydration. Would this dehydration compound give a positive Lieberman-Burchard
test? 
- Yes, it would give a positive Lieberman-Burchard test.