Department of Education

Region V
Division of Camarines Sur
Nabua National High School
San Miguel, Nabua, Camarines Sur

LEARNING ACTIVITY SHEETS NO. 3.2

Name of Student: _________________________________ Subject Teacher: ___________________________

Learning Area : _________________________________ Date: ___________________________

EXPERIMENTAL RULES & SAFETY PROTOCOLS ON THE USE OF SOME EXPERIMENTAL UNITS AND
CHEMICALS

I. Introductory Concept:

Experiments are essential and crucial part of any research endeavor. They are done to discover if
something works or to test hypothesis if it is true. In addition to this, experiments can only be valid when
it is ethical, followed standard procedures and techniques and done with highest honesty and integrity.
In this learning activity sheet, you are expected to familiarize with the safety precautions when
working in a science laboratory, proper disposal of hazardous chemical waste, rules on potentially
hazardous biological agents, human participants rules, use of vertebrate animal rule, and hazardous
chemicals activities/devices rule.
To fully utilize this learning activity sheet, you have to read and understand the instructions and
consult your teacher. Enjoy performing the activities.

II. Learning Skills from the MELCs:

Familiarize standard methods & techniques in performing experiments.

III. Activities:

A. Recall
Activity 1
Laboratory Apparatuses

Direction: Complete each sentence to state the functions of each laboratory apparatus.

We use a BEAKER to
We use a TEST TUBE to
We use a GLASS ROD to
We use a PLATFORM BALANCE to
We use BUNSEN BURNER to
We use SPATULA to
We use GRADUATED CYLINDER to
We use a PIPETTE to

B. Study

A standard laboratory is a place where experiments are carried out. Aside from this, tests,
standard procedures and preparations of materials/ chemicals are done here. It is essential that a
researcher is in a safe place while conducting the experiment. That is why, it is important to know the
safety protocols inside a standard laboratory.

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 LABORATORY SAFETY RULES

The science laboratory is a fundamentally a dangerous place, where there are dangerous chemicals
and risky procedures. And so, to avoid accidents in doing experiments one must follow these safety rules:

1. Follow instructions at all times.

2. Know the location of safety equipment as well as the exit routes.
3. Dress for the laboratory.
4. Don’t eat or drink in the laboratory.
5. Don’t taste or sniff chemicals
6. Don’t play mad scientist in the laboratory.
7. Dispose laboratory waste properly.
8. Know what to do with laboratory accidents.
9. Leave experiments at the laboratory.
10. Don’t experiment on yourself.

RULES IN HANDLING HAZARDOUS CHEMICALS (Based on the Intel ISEF Guidelines)

Before conducting any experiment, it is a protocol to know the risk, the proper handling chemicals as
well as responsible disposal of wastes.

1. A proper risk assessment of chemicals must be done. It is a must include to review the
following factors;
a. Toxicity c. Flammability
b. Reactivity d. Corrosiveness
2. Ensure that the experiment adhere to the call of environmentally responsible chemistry. Thus,
avoiding the use and production of hazardous substances during chemical process.
The principles to observed are as follows:
a. Waste prevention
b. Use of safest possible chemicals and products
c. Design of least possible hazardous chemical synthesis
d. Use of renewable materials
e. Use of catalysts in order to minimize chemical usage
f. Use solvents and reaction conditions that are safe as possible.
g. Maximization of Energy
h. Minimization of accident potential
3. Always refer to the Chemical’s Safety Data Sheet before using one.
4. Familiarize the waste disposal of chemicals to be used.
5. Obtain proper clearance when buying government-controlled chemicals. E.g. hydrochloric
acid
6. Secure proper permit when buying prescription drugs. Student-researchers are not allowed to
administer prescription drugs to human participants. Also, when administering prescription
drugs to animals, a veterinarian must be present.

PROPER DISPOSAL OF HAZARDOUS CHEMICAL WASTE

Proper and responsible chemical waste disposal is also an essential part of experiment and keeping
a researcher safe from any laboratory accidents. Improper disposal of these wastes may pose greater risk
not only to the researchers doing the experiment but also in the community. Read the following to guide you
on proper waste disposal:

1. Read the Material Safety Data Sheet for each chemical to guide you in storing chemical wastes in
appropriate containers.
2. Segregate chemical wastes by compatibility not alphabetically.
3. Chemical waste containers must be labeled with the following information: Full chemical name and
quantity of the waste. For mixtures, each chemical must be listed. Abbreviations are not allowed;

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 date of waste generation; place of origin; bottle number assigned on corresponding waste sheet;
and the tag or label must have the words; “Hazardous Waste.”
4. You may also read Flinn Chemical Disposal guide online if copies are available.

USE OF VERTIBRATE ANIMALS RULE (Based on the Intel ISEF Guidelines)

The following are the guiding principles and rules for the use of animal subjects. These rules are to
ensure that animals will be treated properly and ethically.

Principles

a. Replace vertebrate animals with invertebrates, lower life forms, tissue/cell culture and /
computer simulations if possible.
b. Reduce the number of animals used without compromising statistical validity.
c. Refine the experimental protocol to minimize pain or distress to the animals.
d. Respect animals and their contribution to research.

As much as possible it is endorsed that student-researchers must conduct a non-animal research

method ad use alternative to animal research. However, when the use of animal is necessary the following
rules must be observed:

Vertebrate animals, as covered by these rules, are defined as:

a. Live, nonhuman vertebrate mammalian embryos or fetuses

b. Tadpoles
c. Bird and reptile eggs starting three days (72 hours) at hatching or birth.
Exception: Because of their delayed cognitive neural development, zebrafish embryos may be
used up to seven days (168 hours) post-fertilization.

Rules

1. Justify why there is a need to use an animal in the research experiment and a complete description
of the animal to be used as well as the detailed description of how the animals will be used.
2. The use of animals must adhere to the existing law and jurisdictions in which research is
performed.
3. There must be NO death of vertebrate animal due to the experimental procedure.
4. Animals must be free from stress.
5. Animal Handling protocols must be observed at all times.
• Animals must be treated kindly and cared for properly.
• Consultation and guidance of an expert (veterinarian) is required.
• Animals must receive proper medical assistance when emergency or illness occurs.
• Disposition of animals must be conducted in a responsible and ethical manner.
• Euthanasia for tissue removal and/or pathological analysis is not permitted for
researches conducted at home/school/ field.
• Livestock or fish raised for food using standard agricultural/aquaculture production
practices may be euthanized by a qualified adult.

POTENTIALLY HAZARDOUS BIOLOGICAL AGENT RISK ASSESSMENT (Based on the Intel ISEF
Guidelines)

Suppose, a study requires the use of a biological agent. Before the researchers start the
experiment, they must assess the risk of the agent to be studied.
Risk assessment defines the potential level of harm, injury or disease to plants, animals and
humans that may occur when working with biological agents. The end result of a risk assessment is the
assignment of a biosafety level which then determines the laboratory facilities, equipment, training, and
supervision required.

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 Biological agents, plant or animal, are classified according to biosafety level risk groups. These
classifications presume ordinary circumstances in the research laboratory, or growth of agents in small
volumes for diagnostic and experimental purposes.
1. BSL-1 risk group contains biological agents that pose low risk to personnel and the
environment. These agents are highly unlikely to cause disease in healthy laboratory workers,
animals or plants. The agents require Biosafety Level 1 containment. Examples of B S L -1
o r g a n i s m s a r e : Agrobacterium tumifaciens, Micrococcus leuteus, Neurospora crassa, Bacillus
subtilis.
2. BSL-2 risk group contains biological agents that pose moderate risk to personnel and the
environment. If exposure occurs in a laboratory situation, the risk of spread is limited and it rarely
would cause infection that would lead to serious disease. Effective treatment and preventive
measures are available in the event that an infection occurs. The agents require Biosafety Level 2
containment. Examples of BSL-2 organisms are: Mycobacterium, Streptococcus pneumonia,
Salmonella choleraesuis.
3. BSL-3 risk group contains biological agents that usually cause serious disease (human, animal or
plant) or that can result in serious economic consequences. Projects in the BSL-3 group are
prohibited.
4. BSL-4 risk group contains biological agents that usually produce very serious disease (human,
animal or plant) that is often untreatable. Projects in the BSL-4 group are prohibited.

Levels of Biological Containment

There are four levels of biological containment (Biosafety Level 1–4). Each level has guidelines for
laboratory facilities, safety equipment and laboratory practices and techniques.

1. BSL-1 containment is normally found in water-testing laboratories, in high schools, and in

colleges teaching introductory microbiology classes. Work is done on an open bench or in an
appropriate biosafety hood. Standard microbiological practices are used when working in the
laboratory. Decontamination can be achieved by treating with chemical disinfectants or by steam
autoclaving. Lab coats and gloves are required. The laboratory work is supervised by an
individual with general training in microbiology or a related science.

2. BSL-2 containment is designed to maximize safety when working with agents of moderate
risk to humans and the environment. Access to the laboratory is restricted. Biological safety
cabinets (Class 2, type A, BSC) must be available. An autoclave should be readily available for
decontaminating waste materials. Lab coats and gloves are required; eye protection and face
shields must also be worn as needed. The laboratory work must be supervised by a scientist who
understands the risk associated with working with the agents involved.

3. BSL-3 containment is required for infectious agents that may cause serious or potentially
lethal diseases as a result of exposure by inhalation. Projects in the BSL-3 group are prohibited.

4. BSL-4 containment is required for dangerous/exotic agents that pose high risk of life-
threatening disease. Projects in the BSL-4 group are prohibited.

Rules for Studies with Potentially Hazardous Biological Agents (PHBA)

Students are allowed to do research projects dealing with potentially hazardous biological agent
however they must meet all the rules specified on local and international guidelines. Following these
rules will assure the safety and welfare of the student researcher.

1. Experiments involving the culturing of PHBA, even BSL-1 organisms, is prohibited in a home
set-up. However, specimens may be collected at home as long as they are immediately
transported to a laboratory with the BSL containment.
2. Research determined to be at Biosafety Level 1 (BSL-1) must be conducted in a BSL-1 or
higher laboratory. The research must be supervised by a trained Designated Supervisor or a

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 Qualified Scientist. The student must be properly trained in standard microbiological
practices.
3. Research determined to be a Biosafety Level 2 (BSL-2) must be conducted in a laboratory
rated BSL-2 or above (commonly limited to a Regulated Research Institution). The research
must be reviewed and approved by the Institutional Biosafety Committee (IBC) if the
Regulated Research Institution requires the review. The research must be supervised by a
Qualified Scientist. For a high school BSL-2 laboratory, the SRC must review and approve.
The research must be supervised by a Qualified Scientist.
4. Students are prohibited from designing or participating in BSL- 3 or BSL-4 Research.
5. The culturing of human or animal waste, including sewage sludge, is considered a BSL-2
study.
6. Naturally-occurring plant pathogens may be studied (not cultured) at home, but may not be
introduced into a home/garden environment.
7. All potentially hazardous biological agents must be properly disposed at the end of
experimentation in accordance with their biosafety level. For BSL 1 or BSL 2 organisms:
Autoclave at 121 degrees Celsius for 20 minutes, use of a 10% bleach solution (1:10 dilution of
domestic bleach), incineration, alkaline hydrolysis, biosafety pick-up and other manufacturer
recommendations are acceptable.
8. Research involving human and/or non-human primate established cell lines and tissue
culture collections (e.g., obtained from the American Type Culture Collection) must be
considered a BSL-1 or BSL-2 level organism as indicated by source information and treated
accordingly.
9. The collection and examination of fresh/frozen tissues or body fluids or meat, meat
byproducts, pasteurized milk or eggs NOT obtained from food stores, restaurants, or packing
houses may contain microorganisms. Because of the increased risk from unknown potentially
hazardous agents, these studies must be considered biosafety level 2 studies conducted in a
BSL-2 laboratory under the supervision of a Qualified Scientist.
10. Human breast milk of unknown origin, unless certified free of HIV and Hepatitis C, and
domestic unpasteurized animal milk are considered BSL-2.
11. All studies involving human or wild animal blood or blood products should be considered at a
minimum a Biosafety level 2 study and must be conducted in a BSL-2 laboratory under the
supervision of a Qualified Scientist. Known BSL-3 or BSL-4 blood is prohibited. Studies
involving domestic animal blood may be considered a BSL-1 level study. Any tissue or
instruments with the potential of containing blood-borne pathogens (e.g. blood, blood products,
tissues that release blood when compressed, blood contaminated instruments) must be
properly disposed after experimentation.
12. Studies of human body fluids, where the sample can be identified with a specific person, must
have IRB review and approval, and informed consent.
13. Any study involving the collection and examination of body fluids which may contain
biological agents belonging to BSL-3 or -4 is prohibited.

EXEMPT STUDIES for pre-approval of the schools’ scientific review committee however,
needs risk assessment.

1. Studies involving protists and archaea.

2. Research using manure for composting, fuel production, or other non-culturing experiments.
3. Studies involving decomposition of vertebrate organisms.
4. Studies with microbial fuel cells.
5. Studies involving baker’s yeast and brewer’s yeast, except in rDNA studies.
6. Studies involving Lactobacillus, Bacillus thuringiensis, nitrogen-fixing, oil-eating, and algae-
eating bacteria introduced into their natural environment. (Not exempt if cultured in a petri
dish environment).
7. Studies involving water or soil microbes not concentrated in media conducive to their
microbial growth.

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 8. Studies of mold growth on food items if the experiment is terminated at the first evidence of
mold.
9. Studies of slime molds and edible mushrooms.
10. Studies involving E.coli k-12 (and other strains of E.coli used solely as food source for C.
elegans) that are performed at school and are not subject to additional rules for recombinant
DNA studies or use of antibiotic resistant organisms.

EXEMPT TISSUES

The following types of tissues do not need to be treated as PHBA:

1. Plant tissue
2. Plant and non-primate established cell lines and tissue culture collections. The source and
catalog number of cultures must be identified.
3. Fresh or frozen meat, meat by-products obtained from food stores, restaurants, or packing
houses and pasteurized milks or eggs.
4. Hair, hooves, nails and feathers.
5. Teeth that have been sterilized to kill blood-borne pathogen.
6. Fossilized tissue or archaeological specimen.
7. Prepared fixed tissue.

C. Activity

Activity 2
BETTER BE SAFE THAN SORRY

Direction: Write the observed laboratory safety rule based on the picture presented.

PICTURE A PICTURE B

Source: National Cancer Institute on Unsplash.com Source: National Cancer Institute on Unsplash.com
https://unsplash.com/photos/rb8hr3cXD4A https://unsplash.com/photos/jeT_DE3esSI
PICTURE C

Source: National Cancer Institute on Unsplash.com https://unsplash.com/photos/MOj06jBit0s

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 Activity 3
PROPER WASTE DISPOSAL

Direction: Read the news article below and then, answer the guide questions.

Man nabbed in Batangas for illegal discharge of hazardous waste

By Marita Moaje
August 20, 2020, 5:52 pm

MANILA – The Department of Environment and Natural Resources (DENR) will file multiple
charges against a man in Batangas who was arrested because of illegal discharge of
hazardous waste from used drums allegedly containing chemicals.
Department of Environment and Natural Resources Undersecretary Benny
Antiporda on Thursday said the arrest was made in coordination with the local government
of Batangas and the Philippine National Police (PNP).
The suspect, identified as Romano Cabrera, is said to be the owner of the lot located
in Barangay Bayudbod, Tuy, Batangas, where used drums containing chemicals used for
making soap are being washed.
Antiporda said the illegal discharge of hazardous waste caused great damage to the
environment.
"Grabe yung ginawang damage nito sa environment wherein naglutangan yung mga
hito at igat sa sapa (grave environmental damages resulted from this incident as catfish and
eels from the nearby stream died)," he said.
A total of 16 used drums were recovered from Cabrera, who is believed to be in the
business of buying and selling of the said drums.
He is now detained at Tuy, Batangas Police Station.
Cabrera will be facing multiple charges which include violation of the Republic Act
9275 or the Philippine Clean Water Act and Republic Act 6969 otherwise known as the
Toxic Substances and Hazardous and Nuclear Wastes Control Act of 1990.
Moreover, the LGU will also be filing charges against Cabrera for lack of necessary
permits from the municipality, including the Mayor’s Permit, to be able to operate his
business.
The DENR’s Environmental Management Bureau is already on-site to check the
water quality in the area and to determine what kind of chemicals were dumped by the
suspect. (PNA)

Electronically retrieved from Philippine News Agency https://www.pna.gov.ph/articles/1112944

Guide questions:

1. What is the existing law in the Philippines governing the proper disposal of hazardous waste?

2. Is the law still relevant especially that we are facing a global pandemic? Why?

3. What do you will happen next after the DENR’s EMB checked the quality of water and found it to
be contaminated by a certain substance?

4. As a student researcher, how will handle the disposal of your own chemical waste? How about your
household medical waste in relation to the pandemic issue?

D. Remember

The success of the experiment also means that the researcher was responsible enough on his
actions. This can only be done by abiding the safety rules, assessing the risk of the experiment
beforehand, ethical on using experimental units and responsible for waste disposal.

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
 E. Additional Task

Direction: Revisit your research problem. Assess the risk and safety measures you will employ in your
study. Use the table below and fill it with necessary information gained from your assessment.

Risk Assessment Safety Measures

Chemical Risks
Biological Risks
Physical Hazards

IV. ASSESSMENT:

Direction. Read the situation carefully and choose the letter of the best answer.

1. What is the benefit of reading the safety data sheet of a chemical?

a. The researcher will know how to safely handle the chemical and its waste disposal.
b. The researcher will know how to safely handle the chemical.
c. The researcher will know how to dispose the chemical.
d. The researcher will know the chemical’s nature.
2. Suppose you are doing an experiment at your school’s laboratory then suddenly you felt hungry,
what is the best thing to do?
a. Eat inside the laboratory.
b. Find a safe spot inside the laboratory to eat.
c. Remove your laboratory gown, clean up, then go out to grab some food.
d. Remove your gloves, wash your hands and invite your co-researcher to eat.
3. The science society encourages student researchers to not use live animals in experiments or at
least minimize the number of animal participation, why is that?
a. Animals have the right to live and be respected.
b. There is a scarcity of experimental animals.
c. The use of experimental animals is unethical.
d. Experimental animals are not a good test unit.
4. Which government agency in the Philippines govern the disposal of hazardous waste?
a. DOH
b. DENR
c. EMB
d. DOE
5. Suppose, you want to culture a BSL-1 agent in your research project, which is the least laboratory
that can accept your experimental unit?
a. BSL-1 facility
b. BSL-2 facility
c. BSL-3 facility
d. BSL-4 facility

V.REFLECTION

What I learned from this Learning Activity

Sheet
What I still need to know about research
design
What I need to clarify

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School
VI. REFERENCES:

Internationa Rules and Guidelines for Science Fairs. https://www.societyforscience.org/intel-isef

Moaje, M. (2020) Man Nabbed in Batangas for Illegal Discharge of Hazardous Waste. Philippine News
Agency https://www.pna.gov.ph/articles/1112944

How to Dispose of Chemical Waste. https://case.edu/ehs/waste-disposal/chemical-waste/how-dispose-

chemical-waste.

Photo sources

National Cancer Institute on Unsplash.com. https://unsplash.com/photos/rb8hr3cXD4A

National Cancer Institute on Unsplash.com. https://unsplash.com/photos/jeT_DE3esSI

Source: National Cancer Institute on Unsplash.com https://unsplash.com/photos/MOj06jBit0s

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LAS 3.2 | Experimental Rules & Safety Protocols on The Use of Some Experimental Units & Chemicals
| Jan Ervin S. Babor | Teacher I | Nabua National High School