SELF-LEARNING PACKAGE IN

BIOTECHNOLOGY
Biological Techniques
Learning Competency

Discuss the ways of preparation of reagents.

Rhoda M. Erdao
Oton NHS, SDO-Iloilo
 Ready to Launch!

Science is a never-ending story. The solution of one mystery is the

seed of many others. In all forms of science like biology or biotechnology,
one thing is common: an ultimate goal “to know.” Curiosity and inquiry
are the driving forces for the development of biotechnology. Scientists and
biotechnologists seek to understand the world and the way it govern. And
usually these begin in a laboratory.
Performing task in a laboratory like preparation of reagents always
starts like a child. A child who needs “to learn to walk before he can run”
is a saying many people probably heard when they were children. The
clear message here is that there are some basic skills one need to master
before he can move on to the next level. And there are plenty of good rea-
sons that following biological techniques, procedures, and methods for
laboratory activities will set one up for success, not the least of which—to
continue the metaphor—is to avoid tripping and falling on your face.

Aim at the Target!

After going through this module, you are expected to:

1. Define reagent.
2. Identify commonly used reagents in biology laboratory.
3. Discuss the preparation of some common chemical reagents .
 Try This!

Give Me that Word!

A word cloud below is an image composed of words related to biological
techniques in laboratory. Based on the presented word cloud, answer
the questions found below the image. Choose your answer from the
words included in the word cloud.

Word Cloud

Questions:
1. In one word: It is a substance or compound added to a system
to cause a chemical reaction or added to test if the chemical
reaction occurs.
2. In one word: It refers to a particular form of procedure.
3. In one word: It is a series of actions conducted in a certain order
of manner.
 Keep This in Mind!

Activity: Select and Match

Name ________________________________________________Score _____________

Year and Section ____________________________________ Date _____________

Activity 4 - Select and Match

Procedure:
1. Match the reagents in Column A with their description in Column B.
2. Write the letter of the correct answer.

Column A Column B

1. Anthrone Reagent A. test for aldehydes, sugars, and

carbohydrates.

2. Benedict’s Qualitative Solution B. test for the presence of simple sugar

3. Iodine Solution (Lugol’s) C. test for protein

4. Biuret Solution D. used for common cellulose assay

and in colorimetric determination

5. Molisch Reagent of carbohydrates.

6. Hanus Solution E. used for starch detection and as a

disinfectant

F. used for the determination of iodine

number

G. used as a qualitative test for the

presence of unsaturation such as

double bonds.
Analysis

1. What is a reagent?
_____________________________________________

___________________________________________________________

____________________________________________________________

2. What reagent(s) is/are used to test carbohydrates? proteins?

___________________________________________________________

___________________________________________________________

____________________________________________________________

3. Why are reagents important in laboratory?

____________________________________________________________

____________________________________________________________

_____________________________________________________________

Abstraction and Generalization

What is a reagent?

A reagent is
a compound or mixture added to a
system to cause a chemical reaction or
test if a reaction occurs. A reagent
may be used to find out whether or
not a specific chemical substance is
present by causing a reaction to occur
Reagents
with it. shuttestock.com
 Preparation of Commonly used Reagents in Biology Lab

1. Anthrone Reagent

• It is used for a common cellulose assay and in

colorimetric determination of carbohydrates.

• Give a green color in dilute and a blue color in

concentrated solutions.

Preparation
1. Weigh 2 g of anthrone and dissolve in Anthrone Reagent
1 liter of concentrated sulfuric acid. tomasci.com

2, Store in a refrigerator.
3. Prepare a fresh solution every 2 day

2. Benedict’s Qualitative Reagent

• Benedict's reagent is a chemical reagent and a complex mixture of

sodium carbonate, sodium citrate and copper (II) sulfate pentahy-
drate.

• Benedict's Test is used to test for simple carbohydrates.

• It can be used to test for the presence of glucose in urine.

Preparation

1. Weigh 173 g of sodium citrate dehy-

drate and 100 g sodium carbonate an-
hydrous and dissolve in 800 mL DI
water .
2. Warm and stir to aid dissolution.
3. Filter if necessary. Benedict’s Qualitative Reagent
labtestinfo.com
4. In a separate container, dissolve 17.3 g copper (II) sulfate pen-
tahydrate in 100 mL DI water. Slowly, while stirring constantly,
add the copper sulfate solution to the first solution.
5. Let cool and dilute to 1 L with DI water.

3. Biuret Solution

• The biuret test, also known as

Piotrowski's test, is test used for
detecting the presence of proteins

• The biuret reaction can be used

to assess the concentration of
proteins because peptide bonds
occur with the same frequency .
per amino acid in the peptide. Biuret Solution
sciencephoto.com

Preparation .

1. Weigh 2.3 g of copper (II) sulfate pentahydrate , then dissolve

in 230 mL of DI water. Set a side.
2. Dissolve 308 g sodium hydroxide in 770 mL of DI water (very
exothermic; cool vessel in an ice water bath) and cool to room
temperature
3. Add all the copper sulfate solution to the sodium hydroxide
solution. Solution should be blue.

4. Hanus solution (for iodine number)

• Chemical name: Hanus solution

• Chemical synonyms: Iodine bromide solution

 • Used to determine iodine values in
oils containing unsaturated organic
compounds

Hanus Solution
atlanticlabo-ics.
5. Iodine Solution (Lugol’s)

• Used as reagent for starch detection

• Lugol's iodine, also known as aqueous

iodine and strong iodine solution.

• It is a solution of potassium iodide with

iodine in water.

• It is a medication and disinfectant used

for a number of purposes.

Preparation

1. Weigh 20 g of potassium iodide and

10 g of iodide.
2. Dissolve 20 g of potassium iodide
in 200 mL of DI water; add 10 g of
iodine; stir to dissolve then dilute
to 1 L.
3. Store in a dark bottle.
Lugol’s Iodine Solution

natural-healthshop.com
 6. Molisch Reagent

• Molisch test is a chemical test

used to detect aldehydes,
sugars, and carbohydrates

• It can be used to differentiate

proteins and amino acids from
carbohydrates. Liquid Molisch Reagent

indiamart.com
Preparation

1. Weigh 5 g 1-naphthol
2. Dissolve 5 g 1-naphthol in 100 mL of 95% ethyl alcohol

Application

My Lab Emotion

The most effective vehicle by which the process of inquiry can be

learned appears to be in a laboratory where the learner experiences,
firsthand, the inquiry process Laboratory is an effective means of
comprehension, understanding , and application of biological
knowledge. All these, go with much responsibility and awareness
while working in the area. The Dos and Don’ts in laboratory should
be mastered to get better result and successful experiment.
The “My Lab Emotion “table shows the Dos (allowed) and Don’ts (not
allowed) while preparing reagents in the laboratory . Answer the table
by putting emoticons: happy face for Dos and

Sad face for Don’ts

 My Lab Emotion Table

Place/Draw a happy face for Dos while preparing reagents

Place/Draw a sad face for Don’ts while preparing reagents

Dos Don’ts

1. Use clean and dry glass apparatus while preparing

the reagents.

2. Follow the specific procedure and amounts/volumes

of chemicals and solvents to be used for the reagent to
be prepared.

3. Dispose any waste paper or any kind of solid waste

anywhere.

4. Never touch, taste, or smell any chemical, whatever

it may be.

5. Never wear a lab-coat and safety goggles inside the

laboratory.
 Reflect

Sharing Thoughts!

Reagents and Coronavirus Testing

Covid-19 Test
youtube.com

Reagent is a fancy word for the main ingredients of any chemical-

based test, which in this pandemic includes inorganic solutions as
well as enzymes, probes, and primers created to match the corona-
virus genome. They are the tip of testing spear in the fight against the
coronavirus, to know if one is positive or negative and this is very im-
portant to the people in the community-local or international. What
will happen if there is a shortage of chemical reagent used to test
coronavirus? How this will affect the individuals and the community?
Please answer the questions on the space provided below or on the
paper.

Answer _____________________________________________________________
_____________________________________________________________
_____________________________________________________________
 Reinforcement & Enrichment

Write It!

Describe the uses of the following reagents commonly used in chemistry.

Write your answer on the paper.

1. Bayer’s reagent
2. Dimethyl glyoxime reagent
3. Fehling’s solution

Assess Your Learning

True or False: Write “T” if the statement is true and “F” if the
statement is false

1. Benedict's Test is used to test for complex carbohydrates.

2. In the preparation of Biuret solution, 2.3 g of copper (II) sulfate

pentahydrate , will be dissolved in 250 mL of DI water.
3. Anthrone reagent must be prepared every five days.
4. To test for aldehydes, iodine solution will be used.
5. When preparing for reagents follow the specific procedure and
amounts/volumes of chemicals and solvents to be used.
6. Hanus solution is used to test if protein is present.
7. Biuret solution is used for detecting the presence of proteins.
8. Excess reagents can be poured back into the stock bottle.
9. Lab coats much be taken off when exiting the lab and entering a non-
laboratory area.

10. Lugol's iodine, also known as aqueous iodine is used to test the
presence of starch.
 References & Photo Credits

http://www.sgtbkhalsadu.ac.in/colleges/dbtevents/1127260320121552021.pdf

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1l.html&psig=AOvVaw08VnPnMpAwzoFaaVC0xHuI&ust=1603123163935000&source=imag
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cCegQIBAA&oq=ad+face&gs_lcp=

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q=solutions+in+laboratory&tbm=isch&ved=2ahUKEwjhzdXc7r3sAhWCHaYKHRNECoUQ2-
cCegQIBAA&oq=solutions+in+laboratory&gs_lcp=CgNpbWcQAzIGCAAQBRAeMgYIABAFEB4
6BwgAELEDEEM6AggAOgQIABBDOgQIABAeOgQIABAYUIP3Fli0shdglr4XaABwAHgAgAHGA
YgBzQ-SAQQwLjE1mAEAoAEBqgELZ3dzLXdpei1pbWfAAQE&sclient=img&ei=xA-MX-
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&ved=0ahUKEwjb7uCkuMDsAhXVBogKHUe8BWoQ_AUIDygB&biw=1366&bih=625&dpr=1

https://www.thoughtco.com/definition-of-reagent-and-examples-605598