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Republic of the Philippines
University of Northern Philippines
COLLEGE OF HEALTH SCIENCES

DEPARTMENT OF MEDICAL LABORATORY SCIENCE

BIOCHEMISTRY :
Activity Manual

FITZ GERALD P. CASTILLO, RMT

Instructor

FITZ GERALD P. CASTILLO, RMT

Presented by:

______________________________________
Student

2018

BIOCHEMISTRY LABORATORY MANUAL 1|Page

 v.2.0
PREFACE

This manual is designed for both clinical laboratory scientists and medical laboratory students
who are studying and interested in learning the important aspects of Biochemistry.

The authors has been tasked to undertake teachings on laboratory aspects of the subject and
that no laboratory manual is yet available and designed for local use. It has commenced that most
materials that should be presented to the students must be simple and easily absorbed.

The manual is presented in a concise manner that special structures are mentioned for the
student to understand well the theoretical concepts of Biochemistry in applying it to actual learning
method.

Constructive criticism and suggestions from knowledgeable colleagues and students are
encouraged, pointing out items that need to be changed, added or deleted for the improvement of
this manual.

FITZ GERALD P. CASTILLO, RMT

FITZ GERALD P. CASTILLO, RMT

BIOCHEMISTRY LABORATORY MANUAL 2|Page

 TABLE OF CONTENTS v.2.0

Front Page 1
Preface 2
Table of Contents 3
Policies, Guidelines and Procedures 4
Special Instructions 6
Activity No. Topic
1 INSTRUMENTATION IN CLINICAL CHEMISTRY 1 LABORATORY 7
2 GENERAL PRINCIPLES OF QUALITY AND SAFETY IN CLINICAL CHEMISTRY 10
3 PATIENT PREPARATION FOR SPECIMEN COLLECTION 12
4 BLOOD SPECIMEN COLLECTION FOR CHEMISTRY ASSAY 13
5 SAMPLE PREPARATION FOR CHEMISTRY ASSAY 18
6 SPECIMEN SAFETY HANDLING AND TRANSPORTATION FOR CHEMISTRY ASSAY 21
7 SPECIMEN REJECTION FOR CHEMISTRY ASSAY 22
8 LABORATORY MATHEMATICS AND SOLUTION PREPARATION 23
9 STUDY OF PIPETTING TECHNIQUES 26
10 CALIBRATION OF PIPETTE 31
11 PREPARATION AND USE OF QUALITY CONTROL CHART
(LEVEY-JENNINGS CHART WITH APPLICATION OF WESTGARD RULES) 34
12 CONSTRUCTING A REFERENCE OR STANDARD CURVE 38

FITZ GERALD P. CASTILLO, RMT

13 SPECTROPHOTOMETRY 40
14 GLUCOSE(HEXOKINASE) DETERMINATION 42
15 CHOLESTEROL DETERMINATION 45
16 TRIGLYCERIDES DETERMINATION 48
17 DIRECT HDL CHOLESTEROL DETERMINATION 51
18 UREA (BUN) DETERMINATION 54
19 CREATININE(KINETIC) DETERMINATION 57
20 URIC ACID DETERMINATION 59

Reference 62

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 v.2.0
POLICIES, GUIDELINES AND EQUIPMENTS

A. Requisition of Materials, Reagents and Equipment

1. Fill-out properly the requisition slips of materials and it should be submitted 2-3 days
before the scheduled performance of the laboratory activity to the College Laboratory
Technician.
2. Renew requisition slips for postponed activity or experiments.
3. Second requisition, loses and breakages incurred during the performance of laboratory
experiments or activities are equally charged to the members of the group or by the
whole class. Performance of experiments or laboratory activities starts from the time the
students get materials up to the time that they return such to the stock room or to the
College Laboratory Technician.
4. Request only the amount of chemicals, reagents, supplies and materials just enough for the
experiment or activity to be performed. Employ microscale or semi-microscale techniques
whenever possible.
5. The student or the instructor will provide some of the materials if it is not available to the
college stock room that is readily available in the commercial markets.

B. Laboratory Activities and Experiments

1. Each group should be composed of 5-10 members depending on the size of the class.
Individual performance is required for small classes.
2. The laboratory instructor should employ strategies to ensure that all students are engaged
during laboratory activities or experiments.
3. The laboratory instructor should conduct pre- and post-laboratory discussion including the
discussion of safety precautions needed in the activity or in the experiment and the
discussion of how wastes should be disposed of properly.
4. The laboratory instructor should discuss other safety measures not included in the safety

FITZ GERALD P. CASTILLO, RMT

policies and important laboratory procedures, operations and techniques specific to the
laboratory subject.
5. Materials borrowed should be returned fifteen (15) minutes before the end of the
laboratory period.
6. Set-up of materials made by any instructor or student should be dismantled by the same
and should dispose of properly the wastes generated from such set-up.

C. Laboratory Safety Policy

1. NO eating, drinking and chewing of gums in the laboratory. Keep water or beverage
container in your bags. None should be seen on the top of the tables and cabinets.
2. All chemicals in the laboratories should be treated dangerous. Never taste any laboratory
material. Smell materials only if instructed to do so. Smell chemicals by carefully fanning
the top of the tube or bottle so that a little of the vapor is directed towards your nose.
3. Know the proper use and location of the safety materials and equipment (fire extinguisher,
sandbox, medicine cabinet, emergency shower, spill cabinet, fire blanker and eye wash
area).
4. Avoid unnecessary noise in the laboratory
5. Students must always stay in their respective groups.
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6. Horseplay and other acts of mischief are strictly forbidden. v.2.0
7. Never leave an ongoing experiment unattended.
8. Work under the fume hood when using fuming and toxic chemicals.
9. NO student is allowed to enter the stock room unless the instructor told you to do so.
10. Only instructors, stockroom personnel and students in the scheduled laboratory and student
with permission are allowed in the laboratories. Casual visitors are NOT allowed in the
laboratories.
11. Report all breakages and loses to the instructor and to the laboratory assistants.
12. Read or check labels on chemical containers.
13. Label all containers.
14. Secure permission from the laboratory instructor whenever experiments have to be
performed outside of the regular class periods.
15. Do NOT perform any experiment alone or without the supervision of the instructor.
16. Report any chemical spill to your instructor. Spill cabinets have been provided and the
materials should be used ONLY for chemical spills.
17. Always know the hazards and the physical and chemical properties of the materials used.
Material Safety Data Sheet (MSDS) are located in the chemistry stockroom and are
available by asking the laboratory assistants.
18. Report to your instructor any accident, MINOR or MAJOR.
19. Follow all written and verbal directions carefully. If you do not understand the directions,
ask your instructor.
20. Return equipment or apparatuses borrowed from the stock room. Arrange tools after use.
21. Use the necessary Personal Protective Equipment (PPE – laboratory gown, hair net, hand
gloves, eye goggles, face mask, etc.).
22. Do NOT wear contact lenses, dangling jewelry and loose clothing. Tie back long hair.
23. Observe good housekeeping practices. Keep working are organized, clean and dry
always.
24. NO using of high-heeled and open shoes in the laboratory.
25. Do NOT apply cosmetics while in the laboratory.
26. Do not use your electronic gadgets inside the laboratory unless the instructor told you to do
so.
27. Put your electronic gadgets into a silent mode to prevent destructions inside the class.

FITZ GERALD P. CASTILLO, RMT

28. Only materials necessary for the activity or experiments should be in the working area.
Books, notebooks and pieces of paper must be kept away from the working area.
29. Throw your wastes in the proper trash bins. All wastes that are not biologically or
chemically contaminated should be surrendered to the laboratory assistants, who shall
place such in the receptacle found in the stock room.
30. Wash hands with soap and water after an experiments.
31. Treat preserved specimens with respect and should be disposed of properly following the
instruction given by your instructor.
32. Turn off faucets, gas tanks and outlets when not in use.
33. If you are the last to use the laboratory , check the following:
a. Faucets, gas tanks and outlets are shut off;
b. Electric fans and lights are put off;
c. Chairs or stools are arranged, and;
d. Windows and doors are closed.

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SPECIAL INSTRUCTIONS: v.2.0

 Use the 1.5” (left) and 0.5” (right, top, and bottom) margin with a RED color.

1 – Drawing
A 1 2 B
2 – Description
1 2

 Follow the margins and borders of your working are as shown above. The selection of
Figure A and B is depends on the special instructions of each activities.
 Scale and put the actual colors of all your drawings.
 In writing, used BLUE ballpoint pen ONLY.
 ALWAYS CITE YOUR REFERENCE WITH THE APA format in all of your activities.
o Bishop, M. L., Fody, E. P. & Schoeff, L. E. (2010). Clinical Chemistry: Techniques, Principles, Correlations. 6th
ed. Lippincott Williams & Wilkins.
o WHO (26 April 2006). Blood Safety and Clinical Technology. Guidelines on Standard Operating
Procedures for Clinical Chemistry. Retrieved on May 12, 2012 from http://www.searo.who.int.
 Group Leaders will collect the activity papers and arrange it alphabetically then

FITZ GERALD P. CASTILLO, RMT

attached the ¼ sheet of paper (write the members and your group Name
alphabetically) with a PAPER CLAMP.

I agree and abide by the set policies, guidelines and additional rules provided in print and
discussed verbally by our instructor and should carry out all tasks with utmost caution. Should
there be instances

Name of the Student (PRINT) _____________________________________

Signature of the Student _____________________________________

Date Signed _____________________________________

BIOCHEMISTRY LABORATORY MANUAL 6|Page

 NAME (SURNAME, FIRST NAME, M. I.) DATE PERFORMED (MM/DD/YYYY) GROUP No. SCORE

COURSE/YEAR/SECTION DATE SUBMITTED (MM/DD/YYYY)

v.2.0

Experiment No. 1
WATER & ITS PROPERTIES

INTRODUCTION:
Solubility in a liquid peculiar property of any substance and it
depends on the chemical makeup of the substance any substance dissolve
in a liquid is said to be soluble in that liquid while the substances which fail
to dissolve in any substance is said to be called insoluble.
Water is the predominant chemical component of living organisms.
Its unique physical properties, which include the ability to solvate a wide range of organic and inorganic
molecules, derive from water’s dipolar structure and exceptional capacity for forming hydrogen bonds.
The manner in which water interacts with a solvated biomolecule influences the structure of each. An
excellent nucleophile, water is a reactant or product in many metabolic reactions. Water has a slight
propensity to dissociate into hydroxide ions and protons. The acidity of aqueous solutions is generally
reported using the logarithmic pH scale. Bicarbonate and other buffers normally maintain the pH of
extracellular fluid between 7.35 and 7.45. Suspected disturbances of acid–base balance are verified
by measuring the pH of arterial blood and the CO2 content of venous blood. Causes of acidosis (blood
pH <7.35) include diabetic ketosis and lactic acidosis. Alkalosis (pH >7.45) may follow vomiting of
acidic gastric contents. Regulation of water balance depends upon hypothalamic mechanisms that
control thirst, on antidiuretic hormone (ADH), on retention or excretion of water by the kidneys, and on
evaporative loss. Nephrogenic diabetes insipidus, which involves the inability to concentrate urine or
adjust to subtle changes in extracellular fluid osmolarity, results from the unresponsiveness of renal
tubular osmoreceptors to ADH.

OBJECTIVES:

FITZ GERALD P. CASTILLO, RMT

At the end of the laboratory session, the student will be able to:
 Determine the properties of water that makes it a suitable medium for sustaining life in biological
systems.
 Determine the properties of water in different solutions.
 Determine the different biomedical importance of water.

MATERIALS:
1) Test tube 11) Sugar
2) Test tube rack 12) Gelatin powder
3) Dropper 13) Copper Sulfate (CuSO4)
4) Beaker 14) Lard (oil)
5) Thistle tube 15) Ethanol
6) Ring Stand 16) Benzene
7) Stirring rod 17) Powdered Citric acid
8) Distilled Water 18) Sodium bicarbonate
9) Starch 19) Silver nitrate (AgNO3)
10) Sodium chloride (NaCl) 20) Dialysis Bag (Pig Intestine)

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PROCEDURE: v.2.0
I. WATER AS UNIVERSAL SOLVENT
1) Get a clean test tube and label it with the solvent to be used.
2) Pour all the test tubes at least 3 mL of water in a test tube.
3) Put your agents to the test tube to test for their solubility.
4) Mix or Stir them equally.
5) In putting the solid particles in a test tube for solubility test, equal amount must be
observed.
6) Observe and note the changes occur during the experiment.
7) Note also the physical texture of the test substances for your references.

Substances Solubility in Water Solubility in Benzene

NaCl
Sugar
Gelatin
CuSO4
Lard (Oil)
Ethanol

II. WATER AS GOOD MEDIUM FOR BIOCHEMICAL REACTIONS

1) Get a clean test tube or any glassware that can hold a minimum of 10mL.
2) On a beaker, mix an equal amount (at least 1mL & 1 gram) of powdered citric acid and
Sodium bicarbonate.
3) Observe for any reaction.
4) After mixing it, try to add a small amount of water to the mixture up to 10mL and observe

FITZ GERALD P. CASTILLO, RMT

for changes.
5) Note the observation.

III. PROPERTIES OF WATER SOLUTIONS

1) Dialysis bag
i. Prepare a ring stand for this experiment.
ii. Make a 1% starch-NaCl solution in a beaker with water.
iii. Put the solution into a dialysis bag. Tie the ends to secure the solution inside the
dialysis bag.
iv. Put / string on it and put it on a water.
v. Make sure that water is in the same with the dialysis bag.
vi. Set a timer for 1 hour and observe what happened.
vii. After 1 hour, Ad. Enumerate the different functions of water in living systems.
viii. Based on your observations, define dialysis and osmosis. Site possible
applications of the above procedures.
ix. A few drops of a dialysate of Silver nitrate on the beaker.
x. A presence of a yellow precipitate indicates the presence of chloride ions that
formed Silver chloride.

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 xi. Observe & record your observations. v.2.0

2) Osmosis
i. Wrap a plastic into a thistle tube it on water and observe what happened.
ii. Check in the level of the water is the same of not.

QUESTIONS FOR RESEARCH:

1) Enumerate the different functions of water in living systems.
2) Based on your observations, define dialysis and osmosis. Site possible applications of the above
procedures.

FITZ GERALD P. CASTILLO, RMT

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