Biochemistry (MC2 L)

Laboratory Activity #1
Common Laboratory Equipment & Procedures

Name:__________________________________ Group:_______
Section:___________ Date:_______________

Welcome to Easter College, Incorporated! Chemistry is the gateway to careers in scientific research in human health.
You may be wondering why you, as a nursing student—or as someone with any major other than chemistry—is required
to take this course. The answer is that all matter is made up of molecules, and chemistry is the science that studies how
molecules behave. If you need to understand matter, you need to know chemistry, and specifically for your academic
course, biochemistry!

I. Biochemistry: An introduction
1. Introduction & Scope of Biochemistry
Biochemistry is the branch of life science which deals with the study of chemical reactions occurring in living cells and
organisms. The term ‘Biochemistry’ was first introduced by the German Chemist Carl Neuberg in 1903. It takes into
account the studies related to the nature of the chemical constituents of living matter, their transformations in biological
systems and the energy changes associated with these transformations. Biochemistry may thus be treated as a discipline
in which biological phenomena are analyzed in terms of chemistry. The branch of Biochemistry for the same reason, it
has been variously named as ‘Biological Chemistry’ or ‘Chemical Biology’. Modern biochemistry has two branches,
descriptive biochemistry and dynamic biochemistry. Descriptive biochemistry deals with the qualitative and quantitative
characterization of the various cell components and the dynamic biochemistry deals with the elucidation of the nature
and the mechanism of the reactions involving these cell components. Many newer disciplines have been emerged from
biochemistry such as enzymology (study of enzymes), endocrinology (study of hormones), clinical biochemistry (study of
diseases), molecular biochemistry (study of biomolecules and their functions) etc. Along with these branches certain
other specialties have also come up such as agricultural biochemistry, pharmacological biochemistry etc. Biochemistry is
related to almost all the life sciences and without biochemistry background and knowledge, a thorough understanding
of health and well-being is not possible. Those who acquire a sound knowledge of biochemistry can tackle the two
central concerns of the biomedical sciences (1) the understanding and maintenance of health, and (2) the understanding
and treatment of diseases.

1.1 Objectives of Biochemistry

The major objective of biochemistry is the complete understanding of all the chemical processes associated with living
cells at the molecular level. To achieve this objective, biochemists have attempted to isolate numerous molecules found
in cells, to determine their structures and to analyze how they function. Biochemical studies have illuminated many
aspects of disease and the study of certain diseases has opened up new therapeutic approaches. In brief the objectives
can be listed as follows:
 Isolation, structural elucidation and the determination of mode of action of biomolecules
 Identification of disease mechanisms
 Study of inborn errors of metabolism
 Study of oncogenes in cancer cells
 The relationship of biochemistry with genetics, physiology, immunology, pharmacology, toxicology etc.

Biochemistry is the study and application of substances, reactions and processes in animals, plants, bacteria and viruses.
Biochemistry has vast scope. Biochemists work in hospitals, industry, drug design and development, environmental
sciences, forestry, agriculture, dietetics, hormone production, vaccine research, virology, immunology, microbiology,
toxicology, food science, plant science and associated areas and in areas from marine biology to entomology not just to
carry out R&D work and develop new products but also to monitor the production, quality and safety of the product.
Biochemists provide diagnostic service, carrying out tests on blood, urine and other body fluids, while researching the
underlying causes of disease and methods of treatment. They find jobs in pharmaceutical and agrochemical companies,
food brewing and biotechnology industry. The postgraduate degree in biochemistry also provides opportunity for
advanced teaching in universities, colleges, medical, dental and veterinary schools and consulting or allied work.
Observational, organizational, computational skills, planning and team work are important for biochemists.

1.2 Importance of Biochemistry in Nursing/ Health Care.

Life is associated with living beings. A cell whether a unicellular cell, like a bacterial cell, yeast cell or a multicellular cell is
capable of self-existence and multiplication, either asexually or sexually. The classical definition describes a cell as living,
as long as it can multiply from one to two. Viruses are accordingly considered as living as they multiply inside a host, but
outside the host, they are non-living, not being able to multiply.
Life is not an oxidation–reduction process in which reduced fuels like carbohydrates, proteins and fats are consumed
and oxidized. Oxidation requires oxygen which we breathe, and in mitochondria, oxygen acts as a terminal acceptor of
electrons from the reduced compounds generating energy for life processes. Thus, each organism undergoes metabolic
processes which are performed by biological catalysts, the enzymes. Though the processes are regulated, the genetic
defects lead to metabolic disorders, which are manifested as disease. The diseases have clinical symptoms. For example,
a defect in the sequence of amino acid in the globin protein of hemoglobin results in ‘sickling’ of the protein and the
disease is known as ‘sickle cell’ anemia.
Biochemistry constitutes the study of chemistry of biomolecules and metabolic processes which constitute and are
essential for life. The nursing profession is concerned with the health of an individual and in order to understand
disease, its manifestation and its diagnostics, a nursing student must understand the metabolic processes of living cell
and their regulation. The identification, diagnostics and characterization of disease, which are based on biochemical
principles, is part of nursing degree curricula. Thus, the study of biochemistry is essential for nursing students and those
practicing nursing.

II. Rules for our Biochemistry laboratory and experiments.

1. Safety Rules:
What are the Safety Do’s and Don’ts for Students?
Life threatening injuries can happen in the laboratory. For that reason, students need to be informed of the correct way
to act and things to do in the laboratory. The following is a safety checklist that can be used as a handout to students to
acquaint them with the safety do’s and don’ts in the laboratory.

Conduct
 Do not engage in practical jokes or boisterous conduct in the laboratory.
 Never run in the laboratory.
 The use of personal audio or video equipment is prohibited in the laboratory.
 The performance of unauthorized experiments is strictly forbidden.
 Do not sit on laboratory tables/ working areas.

General Work Procedure

 Know emergency procedures.
 Never work in the laboratory without the supervision of a teacher.
 Always perform the experiments or work precisely as directed by the teacher.
 Immediately report any spills, accidents, or injuries to a teacher.
 Never leave experiments while in progress.
 Be careful when handling hot glassware and apparatus in the laboratory. Hot glassware looks just like cold
glassware.
 Never point the open end of a test tube containing a substance at yourself or others.
 Never fill a pipette using mouth suction. Always use a pipetting device.
 Make sure no flammable solvents are in the surrounding area when lighting a flame.
 Do not leave lit Bunsen burners unattended.
 Turn off all heating apparatus, gas valves, and water faucets when not in use.
 Do not remove any equipment or chemicals from the laboratory.
 Coats, bags, and other personal items must be stored in designated areas, not on the bench tops or in the aisle
ways.
 Notify your teacher of any sensitivities that you may have to particular chemicals if known.
 Keep the floor clear of all objects (e.g., ice, small objects, and spilled liquids).

Housekeeping
 Keep work area neat and free of any unnecessary objects.
 Thoroughly clean your laboratory work space at the end of the laboratory session.
 Do not block the sink drains with debris.
 Never block access to exits or emergency equipment.
 Inspect all equipment for damage (cracks, defects, etc.) prior to use; do not use damaged equipment.
 Never pour chemical waste into the sink drains or wastebaskets.
 Place chemical waste in appropriately labeled waste containers.
 Properly dispose of broken glassware and other sharp objects (e.g., syringe needles) immediately in designated
containers.
 Properly dispose of weigh boats, gloves, filter paper, and paper towels in the laboratory.

Apparel in the Laboratory

 Always wear appropriate eye protection (i.e., chemical splash goggles) in the laboratory.
 Wear disposable gloves, as provided in the laboratory, when handling hazardous materials. Remove the gloves
before exiting the laboratory.
 Wear a full-length, long-sleeved laboratory coat or chemical-resistant apron.
 Wear shoes that adequately cover the whole foot; low-heeled shoes with non-slip soles are preferable. Do not
wear sandals, open-toed shoes, open-backed shoes, or high-heeled shoes in the laboratory.
  Avoid wearing shirts exposing the torso, shorts, or short skirts; long pants that completely cover the legs are
preferable.
 Secure long hair and loose clothing (especially loose long sleeves, neck ties, or scarves).
 Remove jewelry (especially dangling jewelry).
 Synthetic finger nails are not recommended in the laboratory; they are made of extremely flammable polymers
which can burn to completion and are not easily extinguished.

Hygiene Practices
 Keep your hands away from your face, eyes, mouth, and body while using chemicals.
 Food and drink, open or closed, should never be brought into the laboratory or chemical storage area.
 Never use laboratory glassware for eating or drinking purposes.
 Do not apply cosmetics while in the laboratory or storage area.
 Wash hands after removing gloves, and before leaving the laboratory.
 Remove any protective equipment (i.e., gloves, lab coat or apron, chemical splash goggles) before leaving the
laboratory.

Emergency Procedure
 Know the location of all the exits in the laboratory and building.
 Know the location of the emergency phone.
 Know the location of and know how to operate the following:
a. Fire extinguishers
b. Alarm systems with pull stations
c. Fire blankets
d. Eye washes
e. First-aid kits
f. Deluge safety showers
 In case of an emergency or accident, follow the established emergency plan as explained by the teacher and
evacuate the building via the nearest exit.

Chemical Handling
 Check the label to verify it is the correct substance before using it.
 Wear appropriate chemical resistant gloves before handling chemicals. Gloves are not universally protective
against all chemicals.
 If you transfer chemicals from their original containers, label chemical containers as to the contents,
concentration, hazard, date, and your initials.
 Always use a spatula or scoopula to remove a solid reagent from a container.
 Do not directly touch any chemical with your hands.
 Never use a metal spatula when working with peroxides. Metals will decompose explosively with peroxides.
 Hold containers away from the body when transferring a chemical or solution from one container to another.
 Use a hot water bath to heat flammable liquids. Never heat directly with a flame.
 Add concentrated acid to water slowly. Never add water to a concentrated acid.
 Weigh out or remove only the amount of chemical you will need. Do not return the excess to its original
container, but properly dispose of it in the appropriate waste container.
 Never touch, taste, or smell any reagents.
 Never place the container directly under your nose and inhale the vapors.
 Never mix or use chemicals not called for in the laboratory exercise.
 Clean up all spills properly and promptly as instructed by the teacher.
 Dispose of chemicals as instructed by the teacher.
 When transporting chemicals (especially 250 mL or more), place the immediate container in a secondary
container or bucket (rubber, metal or plastic) designed to be carried and large enough to hold the entire
contents of the chemical.
 Never handle bottles that are wet or too heavy for you.
 Use equipment (glassware, Bunsen burner, etc.) in the correct way, as indicated by the teacher.

Chemical Waste
- All containers used for chemical waste should be labeled with:
 “WASTE” or “HAZARDOUS WASTE”
 Chemical name (as it appears on the MSDS)
 Accumulation start date
 Hazard(s) associated with the chemical waste

Storage Don’ts
 Do not place heavy materials, liquid chemicals, and large containers on high shelves.
 Do not store chemicals on tops of cabinets.
  Do not store chemicals on the floor, even temporarily.
 Do not store items on bench tops and in laboratory chemical hoods, except when in use.
 Do not store chemicals on shelves above eye level.
 Do not store chemicals with food and drink.
 Do not store chemicals in personal staff refrigerators, even temporarily.
 Do not expose stored chemicals to direct heat or sunlight, or highly variable temperatures.

Proper Use of Chemical Storage Containers

 Never use food containers for chemical storage.
 Make sure all containers are properly closed.
 After each use, carefully wipe down the outside of the container with a paper towel before returning it to the
storage area. Properly dispose of the paper towel after use.

2. Material Safety Data Sheets

The procedures in this biochemistry lab manual is
designed and described with an emphasis on safety.
However, no amount of planning or pretesting of
procedures substitutes for awareness and common
sense on the part of the student. All chemicals used
in the procedures outlined here must be handled
with care and respect. The use of chemicals in all
workplaces, including academic research and
teaching labs, is regulated by the government’s
Hazard Communication Standard, a document
written by the Occupational Safety and Health
Administration (OSHA).
Specifically, the OSHA standard requires all
workplaces where chemicals are used to do the
following: (1) develop a written hazard
communication program; (2) maintain files of
Material Safety Data Sheets (MSDS) on all chemicals
used in that workplace (an MSDS is a detailed
description of the properties of a chemical substance,
the potential health hazards, and the safety
precautions that must be taken when handling it); (3)
label all chemicals with information regarding
hazardous properties and procedures for handling;
and (4) train employees in the proper use of these
chemicals. Several states have passed “right-to-
know” legislation that amends and expands the
federal OSHA standard. If you have an interest in or
concern about any chemical used in the laboratory,
the MSDS may be obtained from your instructor or
laboratory director or from the Internet (www.sigma-
aldrich.com, for example). The actual form of an
MSDS for a chemical may vary, but certain specific
information must be present. Figure 1.1 is a partial
copy of the MSDS for glacial acetic acid, a reagent
often used in biochemical research. All chemical
reagent bottles in a workplace, lab, or stockroom
must be labeled to identify potential hazardous
materials and to specify personal protection FIGURE 1.1
necessary for handling. One standard hazard Partial MSDS for glacial acetic acid. MSDS information is
communication system used for this purpose is the subject to change. For the most current version, contact
Hazardous Materials Identification System (HMIS® III Sigma-Aldrich. Courtesy of Sigma-Aldrich Corp., St. Louis,
Wall Chart containing an HMIS® III Label in the lower MO; www.sigmaaldrich.com.
left-hand corner is shown in Figure 1.2). The health,
flammability, physical hazard, and personal FIGURE 1.2
protection codes for the chemical reagent are HMIS® III Wall Chart containing an HMIS® III label (lower
summarized on the bottle label for quick left-hand corner) for glacial acetic acid displaying the
hazard indices for the chemical. HMIS® III is a registered
trademark of the National Paint & Coatings Association, Inc. (NPCA). It is used with permission and may not be further
reproduced. NPCA has granted an exclusive license to produce and distribute HMIS® III materials to J. J. Keller &
Associates, Inc. Those wishing to utilize the HMIS® III system should contact J. J. Keller at 1-800-327-6868 or
www.jjkeller.com.

III. Laboratory Safety Signs/Labels and Personal Protective Equipment.

1. Common Safety Symbols

Hazards in the Lab

 Severe Toxicity - adverse effects of a substance that result either from a single exposure or from multiple
exposures in a short space of time (usually less than 24 hours)
 Irritant - causes redness, inflammation
 Corrosive - "eats away" tissue gradually
 Carcinogenic - causes cancer
 Flammable - easily sets on fire
 Biohazard - substances, materials that pose a threat to the health of living organisms, primarily that of humans.

2. Personal Protective Equipment

Personal Protective Equipment or PPE refers collectively to equipment such as safety glasses, goggles, aprons, lab coats,
gloves, protective shoes, respiratory protective equipment, ear defenders and similar equipment used to protect the
person during their work.

IV. Common Laboratory Apparatus or Materials Used.

Below are the commonly used biochemistry laboratory materials, equipment or apparatus:
Your task for your first experiment is to draw in a long coupon bond (with 1 cm margin on all sides) the common
biochemistry equipment, apparatus or materials used and must state therein the use of such equipment.