Lab Manual Csu

Name: ___________________________________________________ Section: _____________
Laboratory Exercise Title Scores
1 Laboratory Safety and Regulations
Preparation of Solutions and Standardization of

2
Solutions
3A Measuring small volumes using serologic pipets
3B Measuring small volumes using Automated Pipets
4 Venipuncture
5 Preparation of Blood Samples
6 Preparation of Protein Free Filtrate
Plotting of Quality Control Chart Using Levey-

7
Jennings
8 Spectrophotometer
9 Glucose Determination
10 Total Protein Determination
11 Albumin Determination
12 Blood Urea Nitrogen Determination
13 Blood Uric Acid Determination
14 Creatinine Determination
15 Lipid Determination (Triglycerides and Cholesterol)
INTRODUCTION
This course deals with the Basic Principles of Clinical Chemistry, Instrumentation, Automation and
different clinical analytes, Carbohydrates, Lipids, Non Protein Nitrogen and Proteins assays. The student
CLINICAL CHEMISTRY I 18
will be able to understand the basic principles of spectrophometry which are applied in most of the assays
to be undertaken. Aspects of quality control, proper pipetting techniques, proper techniques of specimen
collection and processing are expected to be learned, where skills and competencies will be acquired by
the student and which are applied in the various methodologies. More diligence in research for write up
questions is exhorted, such as those for other method and clinical analytes.
The methodology or procedures in some of these assays may vary according to the reagent kits
available for study; thus, appropriate revisions should be made.
Likewise, more materials may become available in the near future; thus additions to the kinds of
analytes for the study in the experiment are expected.
Safe handling of blood and serum samples is given adequate attention in the instruction, and a
precaution that these are potential sources of infection is emphasized. The student is required to be aware
of these from the start of sample collection, processing, testing, and disposal. Proper waste disposal must
always be observed during the performance of the assays.
Proper behaviour and work ethics is inculcated in the student while in the laboratory. Metal focus
and reserved actions are required in every one’s discipline. Appropriate management and efficient use of
time is to be monitored.
While in the laboratory class, the student is made aware of the dangers and risks that he / she is
exposed to. The following rules and regulations must be strictly observed.
1. EATING / DRINKING in the laboratory is prohibited.
2. Never begin the experiment unless the entire procedures have been read and clearly understood.
The student is required to have full attention during pre-laboratory instructions.
3. Wearing a laboratory gown is mandatory to protect oneself from chemical or specimen splatters.
4. Pipettes assigned for each sample and for every chemicals reagent should be appropriately used;
interchanging pipettes will cause serious contaminations.
5. Electrical equipment and connections should not be handled with wet hands.
6. Avoid spillage of samples and chemicals. If accidents happen, immediately the spill should be
wiped off. Blood and serum spill should be decontaminated.
7. After drawing an amount of the reagent, the cover should be securely replaced on the bottle. The
reagent should never be left uncovered.
8. Bottles containing the reagents should not grasped by the neck but firmly around the body.
9. Needles or lancets or broken tubes should be discarded into appropriate receptacles.
(Technicians room). These should never be thrown into the waste can.
10. Blood clots, serum samples are disinfected and then discarded into rest room bowls. Likewise,
bloodied cotton balls and applicator sticks must be disposed off properly.
11. Test tubes, cuvettes and pipettes must be thoroughly washed before returning them to the
laboratory custodians.
12. Unused samples may be saved. If so, samples should be properly labelled and covered to be
kept in the refrigerator.
13. Spinning of samples in the centrifuges must never be left unattended.
14. Due care for the laboratory glassware and instrument should be exercised in order to prevent
losses, breakage and breakdown.
15. Immediately report untoward accidents or events that may occur to the Instructor.
The student shall have demonstrated attainment of the general course competencies if he / she earn
a grade of at least 85% from the following criteria:
a. Participation in experiment ( Student Performance Checklist)
b. Laboratory reports/ write ups
c. Quizzes and practical examination
d. Term examinations
The grading of laboratory reports / activities shall include:
a. Procedural details/ flow chart illustration/ correct pictures
b. Correct color reactions ( determination of specific analytes )
c. Correct calculation of results and interpretation of values
d. Knowledge and understanding of the theory and principles of the methods used.
e. Write ups of side questions
Laboratory Exercise I
LABORATORY SAFETY AND REGULATIONS
Objectives:
At the end of the activity, the students will be able to:
1. demonstrate the correct procedure for performing the routine hand hygiene
2. demonstrate the procedures for donning and removing personal protective equipment
3. recognize different laboratory safety and hazard symbols
Materials:
1. Personal Protective Equipments
2. Laboratory Hazard Logos Chart
Procedure:
The instructor facilitates in discussing the Safety Measures in Clinical Laboratory, discuss
different uses of Personal Protective Equipment, and introduce to the students the laboratory
hazards
Drawing:
1. Draw the NFPA Hazardous material symbol and identify each classification
2. Draw the Radioactive Hazard Symbol
3. Draw the Biologic Hazard Symbol
QUESTIONS
1. Enumerate the Classification of Fire, its Hazards and the type of Extinguisher used.
2. What is Material Safety Data Sheet? Give at least 10 common hazardous materials and give its
safety information.
3. Give the categories and characteristics of Bioterrorism agents and give examples of each and the
diseases it cause
4. What is the Universal Safety Precaution Statement?
Laboratory Exercise IIA
MEASURING SMALL VOLUMES USING SEROLOGICAL PIPETS
Introduction
In a laboratory setting, volumes are frequently measured. The equipment that is used depends on
the volume needed. Large volumes over 10mL are typically measured with graduated cylinder. Volumes
between 1 and 10mL are often measured with Serological pipet. These pipets are named according to the
maximum volume they can measure, thus a 10mL pipet can measure up to 10mL. It takes practice to learn
how to accurately use a serologic pipet. The standard practice is to allow no more than 10% deviation
from the intended value, although some applications require much less than that.
In this activity, you will be introduced to techniques for pipetting. In the end, you will be asked to
master the pipetting of given volumes to be assessed by your instructor.
Objective
1. differentiate the types and classification of pipets
2. master the proper technique in pipetting
3. identify the errors enclosed in pipetting technique
Materials
1. Pipets
- Serologic (10mL, 5mL, 1mL)
- Automatic Pipet (10mL, 5mL, 1mL)
- Automatic Micropipet (1000uL, 500uL, 50uL)
2. Rubber aspirator
3. Cloth or tissue
4. Liquid reagents (distilled/tap water)
5. Food color
6. Beaker/volumetric Flask (50mL and 100mL)
7. Test tube (5mL, 3mL)
8. Ruler
Pipet Precautions
- Always wear the appropriate PPE
- Use a pipet pump to withdraw and dispense equipment (do not mouth pipet)
- Hold the bottom of the pipet pump when inserting and removing the pipet
- Always keep the pipet in an almost vertical position when there is fluid in the tip
- Use your thumb to roll the pipetting gear up and down. Do not pull or push on the top of the
pipet pump
- Do not pipet a liquid sample into another liquid, unless directed to do so
Procedure
1. Review the Standard Operating Procedures for pipetting found below
2. Label four glass tubes 1-4
3. Using standard pipetting techniques, measure the appropriate volumes of the dye solutions
into the tubes according to the chart below. Be sure to use the correct pipets and correct pipet
pumps.
Tubes Red Green Blue Yellow Total volume Actual % error

expected volume
Tube 1 6.3mL 0.5mL 0.25mL -----
Tube 2 2.4mL 1.08mL ---- 0.73mL
Tube 3 4.0 mL ---- 0.4mL 1.5mL
Tube 4 ---- 2.0mL 0.2mL 1.1 mL
4. After mixing the different dye solutions in the tubes, calculate the expected total volume in
each tube
5. Using a 10mL pipet, measure the volume of the mixed solution in each of the tubes and
record your results in the table
6. Calculate the percent error of your measurements in each tube by using the formula:
% error = expected volume – actual volume x 100

Expected volume
Standard Operating Procedure for Pipetting

A. Choice of correct pipet and pump
1. Choosing the correct pipet
i. For volumes less than 0.2mL, do not use serological pipet. Use a P-1000
micropipet instead
ii. For volumes between0.2mL and 1.0mL, use 1mL serological pipet
iii. For volumes between 1.0mL and 2mL, use 2mL serological pipet
iv. For volumes between 2.0mL and 5mL, use 5mL serological pipet
v. For volumes between 5.0mL and 10mL, use 10mL serological pipet
vi. For volumes greater than 10mL, use graduated cylinder
vii. The smaller the pipet, the more accurate the measurement, so choose the smallest
if there is any choice
2. Choosing the correct pipet pump
i. When using the 1.0 or 2.0 pipets, use the smaller, blue pipet pump
ii. When using the 5.0 and10.0 pipets, use the larger, green pipet pump
B. Assembling the pipet and pump
1. Hold the white bottom of the pipet pump while inserting the large bore end of the pipet into
the hole
2. Insert the pipet until it is surely fastened in the pipet pump, but not so far as to damage the
vacuum chamber. The proper depth is to the cotton plug, if present
C. Removing the liquid with the pipet
1. Place the tip of the pipet into the solution and keep the tip under the surface of the liquid at
all times when removing the liquid. Be careful the liquid does not overflow the container due
to displacement
2. With the tip of the pipet under the surface of the liquid hold the pipet and container at eye
level, gently roll the pump wheel up to pull the solution up into the pipet
3. Pull the solution up until the bottom of the meniscus is at the volume value desired
4. Never lay the pipet down with liquid in it. Never allow the liquid to reach the cotton plug or
to enter the pump. Alert the instructor if it happens.
D. Dispensing the liquid
1. Move the pipet into the recipient container. Roll the pump wheel down(all the way) to
dispense the solution from the pipet
2. Touch the tip of the pipet to the side of the container so that the adhesion pulls on any liquid
on the side of the pipet
3. Allow the solution to leave the pipet, but do not force out the last tiny bit. Pipets labelled TD
manufactured allow this last drop to remain in the pipet without affecting measurements.
4. When there is not a specific order required to dispensing solutions in the container, always
add the smallest volume first and then the larger volumes progressively.
E. Removing the pipet from the pump
1. Holding onto the bottom of the pump, gently twist and pull the pipet out of the pump
2. Discard the pipet if it is disposable or place it in the cleaning tank if it is not
Student Performance Checklist
Evaluation of Pipetting Techniques
Rating system
2 = Satisfactorily Performed 1 = Needs Improvement 0 = Incorrect/Did not perform
2 1 0
1. Systematic and organized preparation of materials needed
2. Correct selection of the type of pipette to be used
3. Correct assembly of the pipet pump
4. Correct measurement of the required volumes
5. Correct dispensing of liquid
6. Correct removal of pipet pump
7. Cleanliness and orderliness of area while
Score
Laboratory Waste Disposal
Non-hazardous liquid, for the most part, can be poured down the drain. Non-hazardous solid waste can
usually be put into the garbage, taking into consideration the proper segregation of waste.
LABORATORY REPORT
Laboratory Exercise IIB
Name: ___________________________________________________
MEASURING VERY SMALL VOLUMES WITH A MICROPIPET Date: ______________
Course/Year/Section/Set: __________________________________
Group No: ______________
Introduction
Clinical Laboratory often works at the micro-scale. The prefix micro means 10 -6. Therefore,
Results and
microliter (µl)
Observation
is one-millionth of a liter or 1/1000 of a millimetre. To measure this small of a volume, a
Paste the
digital micropipette
picture of your
is needed.
results on
Micropipettes
the box provided
are name by their maximum capacity so a P10 measures
from 1-10 µl, a P100 measures from 10-100 µl, and a P1000 measures from 100 – 1000 µl.
In this activity, you will learn how to use the micropipets to help you learn a “secret code”.
Micropipetting
Tube 1 PrecautionsTube 2 Tube 3 Tube 4
a. The adjustment release button next to the display button must be depressed for adjusting
volume
b. Do not adjust below or over the volumes stamped on the pipette under the display
window
CLINICAL CHEMISTRY I
c. Only aspirate liquids when a tip in laced on the tip holder
d. Always keep the pipette in a vertical position when the liquid is in the tip
e. Slowly and smoothly operate the button plunger when aspirating and dispensing liquids
Procedures
Aspiration: Immerse the pipette tip in the liquid. Allow the plunger to move up smoothly to
the rest position. Wait a second so that the liquid has time to move up into the tip.
Distribution: place the pipette tip at an angle of 10 to 45 degrees against the inside wall of
the receiving vessel. Depress the plunger smoothly to the first stop position.
Purge: wait one second, and then depress the plunger to the second stop position (blow-out
stroke) to remove any remaining sample from the tip
Home: allow the plunger to move up to the rest position. Eject the tip.
Sample Activity
1. Get a clean 96-well assay plate and identify the numbered columns and lettered rows
2. Using the chart below, add the appropriate volumes of each of the basic dye color
solutions to the appropriate wells. Note that the wells may only have one color, multiple
colors or none at all. Also note that the units are given in either µl or mL
3. When complete, exam the plate and determine the secret code
Pipet the following amount s in the designated assay plate cells

Cell Yellow Red Blue Green Cell Yellow Red Blue Green
D4 0 0 350µl 0 C7 50 µl 50 µl 50 µl 200 µl
E4 0 0 350 µl 0 D7 50 µl 50 µl 50 µl 200 µl
F4 350 µl F7 50 µl 50 µl 50 µl 200 µl
C8 0.2mL 50 µl 100 µl C6 0.1 mL 50 µl 50 µl 150 µl
F8 0.2mL 50 µl 100 µl E6 0.1 mL 50 µl 50 µl 150 µl
A12 300 µl 50 µl F6 0.1 mL 50 µl 50 µl 150 µl
G12 300 µl 50 µl C5 50 µl 100µl 200µl 0
B11 200 µl 100 µl 50 µl F5 50 µl 100µl 200µl 0
C11 200 µl 100 µl 50 µl C3 0 0 300µl 50 µl
D11 225 µl 100 µl 50 µl F3 0 0 300µl 50 µl
E11 200 µl 100 µl 50 µl C2 0 50 µl 250µl 50 µl
F11 200 µl 100 µl 50 µl F2 0 50 µl 250µl 50 µl
C10 50 µl 0.25mL 50 µl D1 0 100µl 250µl 0
E10 50 µl 0.25mL 50 µl E1 0 100µl 250µl 0
D9 350 µl F1 0 100µl 250µl 0
Evaluation of Pipetting Techniques
Rating system
2 1 0
2. Correct selection of the type of pipette to be used
3. Correct assembly of the pipet pump
4. Correct measurement of the required volumes
5. Correct dispensing of liquid
6. Correct removal of pipet pump
Score
Non-hazardous liquid, for the most part, can be poured down the drain. Non-hazardous solid waste can
usually be put into the garbage, taking into consideration the proper segregation of waste.
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________

Group No: ______________
Results and Observation

Paste a picture of your results on the box provided
Remarks
_______________________________________________________________
_______________________________________________________________
_______________________________________________________________
_______________________________________________________________
_______________________________________________________________
Post
_______________________________________________________________
Laboratory
Questions _______________________________________________________________
1. _______________________________________________________________ What
_______________________________________________________________ are the
two
general types of micropipette? Differentiate the two.
2. State the reason why the thumb finger is not used in controlling the flow of liquid in the
pipette.
3. What are the factors affecting pipetting performance
Laboratory Exercise III

PREPARATION AND STANDARDIZATION OF SOLUTIONS
Introduction
A Solution is a homogenous mixture which is made up of two or more substances. In an aqueous
solution, the water is usually considered as the solvent or medium in which the solute is dissolved. The
concentration of a solution refers to the amount of solute dissolves in a unit amount of solvent. The
determination of the concentration of a solution is important in chemistry.
Objectives
1. learn to express concentration of solutions
2. learn the basic principles in the preparation and standardization of solutions
Materials
Apparatus
Analytical Balance Burner
Burette Tripod
Timer Thermometer
Glassware
Erlenmeyer Flask Watch glass
50 mL Volumetric Flask Graduated Cylinder
Spatula Beaker
250 mL Volumetric Flask amber bottle
Chemicals
Potassium and acid phthalate Methyl orange
Sodium carbonate Phenolphthalein
Hydrochloric Acid Sodium Hydroxide
Distilled water
Procedures
I. Preparation of 1N HCl
a. Half fill the 50mL volumetric flask with distilled water
b. Gently place 4.17 mL concentrated HCl into the flask
c. Fill the flask with distilled water up to the mark. (This should be done with caution.
Remember the general rule: add Acid to Water)
d. Mix well by inversion and transfer to a clean glass container.
e. The prepared solution is 1N HCl
II. Preparation of 1N NaOH
a. Boil about 200mL of distilled water for 5 minutes to remove carbon dioxide. Allow to
cool, cover with a watch glass, and transfer while still warm (about 40C) to a 250mL
volumetric glassware
b. Weigh 2g of NAOH pellets
c. Place the sodium hydroxide pellets into a half-filled 50 mL volumetric flask with boiled
distilled water
d. Fill with boiled distilled water up to the mark
e. Mix well by inversion and transfer to a clean amber bottle
III. Standardization of Solution
a. Standardization of HCl against Sodium Carbonate
i. Dry 0.2g sample of primary standard sodium carbonate at 110C and dissolve into
50mL distilled water in an Erlenmeyer Flask
ii. Add 1-2 drops of modified Methyl Orange
iii. Prepare 0.1N HCl from previously prepared 1N HCl and place into an acid
burette
iv. Titrate the sodium carbonate solution against 0.1N HCl, taking an end point as
red to red-orange color
v. Calculate the volume of HCl of the acid using the equivalent weight of 53.0 for
sodium carbonate
b. Standardization of NaOH against Potassium and Acid Phthalate
i. Dry the 1260 mg sample of primary standard potassium acid phthalate at room
temperature and dissolve in 50mL of distilled water in an Erlenmeyer flask.
ii. Add 2-3 drops of phenolphthalein
iii. Prepare 0.1N NaOH from previously prepared 1N NaOH and place to a base
burette
iv. Titrate the potassium acid phthalate against 0.1N NaOH, taking the end-point as
white to faint pink
v. Calculate the normality of the base using the equivalent weight of 204.22 for
potassium acid phthalate
Rating system
2 = Satisfactorily Performed 1 = Needs Improvement 0 = Incorrect/ Did not perform
2 1 0
2. Correct preparation of Acid
3. Correct preparation of Base
4. Correct set up of acid / base burette
5. Correct observation of end-point
Score
A. Neutralization
If liquids meet all standards for the sanitary sewer except for pH, campus employees may
neutralize the solution before pouring down the drain. Use proper equipment. Goggles, gloves,
apron, and hood are required. Add neutralizing agent slowly, stirring constantly.
Acidic solutions ( pH <5)

Adjust the pH to 5-9 using a dilute solution (e.g. KOH, NaOH, NaHCO3). Use a pH
meter, indicator solution, or pH paper to determine the pH.
Flush down the drain of a chemical sink with 20 volumes of cool water.
Basic solutions ( pH > 9)
Adjust pH to 5-9 using a dilute solution (e.g. HCl, H2SO4, HNO3 ). Use a pH meter,
indicator solution, or pH paper to determine pH.
Note: For highly concentrated acids, neutralization with a relatively dilute basic solution will take
a very large volume of base and a long time. In this case, consider neutralization using a
concentrated basic solution with plenty of ice for an ice bath, performed slowly and carefully and
with constant stirring. Monitor the temperature of the solution with a suitable thermometer to
ensure that the solution doesn't get too hot. The same is true for neutralizing some concentrated
bases.
B. Safely Sewered
Small amounts of this material may be suitable for sanitary sewer or trash disposal. Sodium
carbonate and Potassium acid phthalate can be flushed down the drain.
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________
Results and Observations
Preparation of 0.1N HCl

Observation:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Color of Solution before titration:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Color of solution when the end point is reached:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Preparation of 0.1N NaOH
Observation:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Color of Solution before titration:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Color of solution when the end point is reached:
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Complete the table
0.1N HCl 0.1N NaOH
Initial Volume
Final Volume
Calculation
Show the computation for the Standardization of HCl
Show the computation for the standardization of NaOH
Problem Solving
1. What is the normality of a 200mL solution containing 5grams of H2SO4?
2. How much NaCl must be dissolved to prepare 500mL of 5 molar solutions?
3. How many mL of 8N H2SO4 will neutralize 45mL of 6N KOH?
4. Solve the equivalent weight of H2CO3?
5. Compute for the molarity of the following:
a.10N H2SO4
b. 8M HCl
Laboratory Exercise IV
VENIPUNCTURE
Introduction
Venipuncture is the collection of a blood sample from a vein for laboratory testing. Venous blood
is the specimen of choice for most routine laboratory tests. It is deoxygenated, dark red blood. The
venipuncture procedure is complex, requiring both knowledge and skill. The three common venipuncture
methods include syringe, evacuated tube system and butterfly method.
The syringe method is used when small amounts of blood are needed or when the patient has
small or fragile veins. The vacuum tube method is often utilized for routine blood draws and is good for
collecting multiple vials of blood. The butterfly method is more appropriate for patients with veins that
are difficult to access.
Objectives
At the end of the activity, the student shall be able to:
1. differentiate among the various needle sizes as to gauge, length and purpose
2. describe the OSHA – required safety needles and equipment
3. discuss methods to dispose of contaminated needles safely
4. differentiate among an evacuated tube system, a syringe, and a winged blood collection set,
and state the advantage and disadvantage of each
5. list the proper drawing order for collection tubes
6. identify the additive or preservative if present
Materials
1. evacuated tube system collection set
2. syringe
3. winged blood collection set
Procedure
I. Tourniquet Application
1. Position the vinyl or latex strip 3 to 4 inches above the venipuncture site. Avoid areas with a
skin lesion or apply the tourniquet over the patients gown
2. Grasp both sides of the toniquet and, while maintaining tension, cross the tourniquet over the
patients arm
3. Hold both ends between the thumb and forefinger of one hand close to the arm
4. Tuck a potion of the left side under the right side to make a partial loop facing the antecubital
fossa
5. A properly applied tourniquet will have the ends positioning up away from the venipuncture
site
6. Pull the end of the loop to release the tourniquet with one hand. The tourniquet should only
be for one minute.
II. Evacuated Tube System Method
1. Position the patients arm slightly bent in a down ward position so that the tubes fill from the
bottom up. Do not allow blood to touch the stopper – puncturing needle. Do not let the
patient hyperextend the arm. Ask the patient to make a fist.
2. Apply tourniquet 3-4 inches above the antecubital fossa. Palpate the area in a vertical and
horizontal direction to locate a large vein and to determine the depth, direction, and size. The
median cubital vein is the vein of choice followed by the cephalic vein. The basilica vein
should be avoided if possible. Remove the tourniquet and have the patient open his or her
fist.
3. Clean the site with 70% isopropyl alcohol in concentric circles moving outward and allow it
to air dry
4. Assemble the equipment while the alcohol is drying. Attach the multisample needle to the
holder
5. Insert the tube into the holder up to the tube advancement mark
6. Reapply the tourniquet. Do not touch the puncture site with an unclean finger. Ask the
patient to remake a fist. Patient should be instructed not to “pump” or “continuously clench”
the fist to prevent hemoconcentration
7. Remove the plastic needle cap and examine the needle for defects such as nonpointed or
barbed ends
8. Anchor the vein by placing the thumb of the non-dominant hand 1 to 2 inches below the site
and pulling the skin taut
9. Grasp the assembled needle and tube holder using your dominat hand with thumb on the top
near the hub and your other fingers beneath. Smoothly insert the needle into the vein at a 15
– 30 degree angle with the bevel up until you feel a lessening of resistance. Brace the fingers
against the arm to prevent movement of the needle when changing tubes.
10. Using the thumb, advance the tube onto the evacuated tube needle, while the index and the
middle fingers grasp the flared ends of the holder
11. When blood flows into the tube, release the tourniquet and ask the patient to open the fist
12. Gently remove the tube when the blood stops flowing into it. Gently invert anticoagulated
tubes promptly. Insert the next tube using the correct order of draw. Fill tube completely
13. Remove the last tube collected from the holder and gently invert
14. Cover the puncture site with clean gauze. Remove the needle smoothly and apply pressure or
ask the patient to apply pressure
15. Activate
16. Dispose the needle/holder assembly with the safety device into the sharps container
17. Label the tubes before leaving the patient and verify identification with the patient ID band
or verbally with an outpatient. Observe any special handling procedures. Complete
paperwork
18. Examine the puncture site and apply bandage. Place the bandage over folded gauze for
additional pressure
19. Prepare sample and requisition for transportation to the laboratory. Dispose used supplies
20. Thank the patient, remove gloves and wash hands
III. Syringe Method
1. Performs steps 1 – 3 of venipuncture using Evacuated Tube System
2. Assemble equipment as the alcohol is drying. Attached the hypodermic needle to the syringe.
Pull the plunger back to ensure that it moves freely and then push it forward to remove any
air in the syringe
3. Reapply the tourniquet, remove the needle cap and inspect the needle
4. Ask the patient to remake a fist, and anchor the vein by placing the thumb of the non-
dominant hand 1-2 inches below the site and pulling the skin taut
5. Hold the syringe in the dominant hand with the thumb on top near the hub and the other
fingers underneath. Smoothly insert the needle into the vein at 15-30 degree angle with the
bevel up until you feel a lessening resistance. A flash of blood will appear in the syringe hub
when the vein has been entered. Brace the fingers against the arm to prevent movement of
the needle when pulling back on the plunger
6. Pull back the syringe plunger slowly using the non-dominant hand to collect the appropriate
amount of blood
7. Release the tourniquet and have the patient open the fist
8. Cover the puncture site with gauze, remove the needle smoothly, activate the safety shield,
and apply pressure
9. Remove the needle from the syringe and discard it in the sharps container
10. Attached the blood transfer device to the syringe
11. Holding the syringe vertically with the blood transfer device at the bottom, advance the
evacuated tube onto the internal needle in the blood transfer device. Tubes will fill the
vacuum in the tube. Keep the tube in a vertical position to ensure that the tubes fill the
bottom up to avoid cross-contamination. Do not push on the plunger
12. Fill the tubes in correct order. Mix anticoagulated tubes as soon as they are removed from
the transfer device
13. /after tubes are filled, the entire syringe and blood transfer are discarded into a sharps
container
14. Label the tubes and confirm identification with the patient
15. Examine the puncture site and apply bandage
Evaluation of Tourniquet Application and Vein Selection Competency
Rating system
2 1 0
1. Positions arm correctly for vein selection
2. Selects appropriate tourniquet application site
3. Places tourniquet in flat position behind arm
4. Fastens tourniquet at appropriate tightness
5. Loop and loose end do not interfere with puncture site
6. Asks patient to clench fist
7. Palpates entire area or both arms if necessary, using right finger
8. Selects antecubital area
9. Removes tourniquet smoothly
10. Remove tourniquet in a timely manner
Score
Evaluation of Venipunture using an Evacuated tube Competency
Rating System
2 = Satisfactorily performed 1 = Needs improvement 0 = incorrect/did not perform
2 1 0
1. Cleanses the site and allows it to dry
2. Assembles equipment
3. Reapplies tourniquet
4. Does not touch puncture site with unclean fingers
5. Removes needle cap and examines the needle
6. Anchors vein below the puncture site
7. Smoothly enters vein at appropriate angle with bevel up
8. Does not move needle when changing tubes
9. Collects tubes in correct order
10. Fills tube completely and mixes promptly
11. Releases tourniquet within one minute
12. Removes last tube collected from the holder
13. Covers puncture site with gauze
14. Removes the needle smoothly and applies pressure
15. Activates any safety feature
16. Disposes of the needle in sharps container with the safety device activated
and attached to the holder
17. Labels tubes
Score
Evaluation of Venipuncture Using a Syringe Competency
Rating System
2 = Satisfactorily Performed 1 = Needs improvement 0 = Incorrect/Did Not perform
2 1 0
1. Cleanses the site and allows it to dry
2. Assembles equipment
3. Reapplies tourniquet
4. Does not touch puncture site with unclean fingers
5. Removes needle cap, examines the needle and check the plunger
movement
6. Anchors vein below the puncture site
7. Smoothly enters vein at appropriate angle with bevel up
8. Does not remove needle when plunger is retracted
9. Collects appropriate amount of blood
10. Releases tourniquet within one minute
11. Covers puncture site with gauze
12. Removes the needle smoothly and applies pressure
13. Use blood transfer device to fill tubes
14. Fills tube in correct order and mixed promptly
15. Disposes of the needle in sharps container with the safety device activated
and attached to the holder
16. Labels tubes
Score
Non-hazardous liquid, for the most part, can be poured down the drain. In this case, non-infectious blood
is drained in the sink.
Non-hazardous solid waste can usually be put into the garbage, taking into consideration the proper
segregation of waste. Place the cotton, tissue, syringe barrel and plunger and anything soiled with
contamination of blood and other body fluids in a yellow (infectious) waste container.
Needles and other sharp objects is placed in a Sharp Container and kept for proper disposal.
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________

Group No: ______________
Complete the table
Table 1. Order of Draw for Syringe Method

Order of Draw Tube Stopper Color Rationale for Collection Order
Table 2. Order of Draw for Evacuated Tube Method

Order of Draw Tube Stopper Color Rationale for Collection Order
Practical Examination
VENIPUNCTURE EQUIPMENT EXERCISE
Instructions: State or assemble (if requested) the appropriate equipment for the situations described in this
activity. Include the number and stopper color of evacuated tubes, needle size, syringe size, or winged blood
collection set, if appropriate. Instructors may specify the inclusion of other supplies.
1. collection of CBC from a 35-year old woman

_______________________________________________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________
_______________________________________________________________________________________
2. collection of CBC from a 3year old boy
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
3. collection of a CBC and electrolytes from a 40-year old woman
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
4. Collection of amylase from the hand of a patient who is taking anticoagulants
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
5. Collection of PT from an elderly patient
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
6. Collection of chemistry profile from patient with latex allergy
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
7. Assemble the equipment to collect a type and crossmatch on a 50 year old man
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
8. Assemble equipment to collect a cardiac risk profile and ESR from a patient with fragile veins
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
9. Assemble the equipment to collect a lead level from a 2 year old patient
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
10. Assemble tubes in the order they would be drawn from a CBC, APTT, and a glucose using an evacuated tube
system
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
Evaluation of Equipment Selection and Assembly Competency
Rating System:
2 1 0
1. Collects all necessary equipment and supplies
2. Selects appropriate tubes for requested test
3. Selects correct number of tubes or syringe size
4. Correctly attaches needle to holder or syringe
5. Does not uncap needle prematurely
6. Advances tube correctly into holder or checks plunger movement
7. Arranges supplies and extra tubes conveniently
Score
Laboratory Exercise V
PREPARATION OF BLOOD SAMPLES
Introduction
Blood consist of fluid portion (plasma) and formed elements. The plasma constitutes about 55%
to 60% volume of the whole blood. The formed elements are the red cells, white blood cells and blood
platelets. Preparation of blood specimen for quantitative analyses of tis constituents is in the form if
serum, plasma, whole blood and protein-free filtrate. If blood is removed from any vessel, it forms clots
or coagulates in a few minutes. The whole mass becomes gelatinous. If left undisturbed, a clear straw-
colored fluid is gradually squeezed out of the whole blood; this is called serum.
Blood plasma can be separated from formed elements an anticoagulant is added immediately after
the blood has been obtained. If it is centrifuged, a fluid that is also clear and straw color is obtained. This
is called plasma.
Since the whole blood is made of formed elements and plasma, it also contains proteins which
must be removed in order not to interfere with colorimetric determination. Such blood sample is treated
with substance to remove protein. The resulting solution is clear filtrate know as protein-free filtrate
(PFF).
Whole blood – formed elements = plasma
Whole blood – (formed elements – clotting factors) = serum
Plasma – protein = protein-free filtrate
Objective
At the end of the activity, the students will be able to
1. prepare and differentiate blood samples serum, plasma and whole blood
2. determine the use of each blood samples
Materials
1. Blood Extraction set
2. Evacuated tubes (Plain tube and Heparin)
3. Applicator stick
4. Centrifuge
5. Timer
6. 5mL test tube
7. Test tube rack
8. Air displacement pipet and disposable tips
Procedure:
1. Using the evacuated tube system of blood collection, draw blood into three (3mL) evacuated
tubes (1 tube without anticoagulant and 2 heparinized tubes)
2. Allow the blood specimen to stand for 10 minutes.
3. Centrifuge the blood specimen (1 without anticoagulant and 1 with anticoagulant) for 10
minutes at 2500 rpm.
4. Aspirate the liquid portion of the blood. Be very careful not to aspirate the red cells and
transfer in a clean test tube labelled as follows:
a. Serum (without anticoagulant)
b. Plasma (with heparin)
5. Label “whole blood” on 1 plain test tube and aspirate 2 mL of whole blood from a
heparinized tube.
Evaluation of Preparation of Blood Samples
Rating System:
2 1 0
1. Collects all necessary equipment and supplies
2. Selects appropriate tubes for requested test
3. Correctly labels tubes
4. Correctly followed the procedure
5. Correctly aspirates blood samples
6. Maintained a neat and clean work area
Score
Non-hazardous liquid, for the most part, can be poured down the drain. In this case, non-infectious blood
is drained in the sink.
Non-hazardous solid waste can usually be put into the garbage, taking into consideration the proper
segregation of waste. Place the cotton, tissue, syringe barrel and plunger and anything soiled with
contamination of blood and other body fluids in a yellow (infectious) waste container.
Laboratory Exercise VI
PREPARATION OF PROTEIN - FREE FILTRATE
Introduction
Blood consist of fluid portion (plasma) and formed elements. The plasma constitutes about 55%
to 60% volume of the whole blood. The formed elements are the red cells, white blood cells and blood
platelets. Preparation of blood specimen for quantitative analyses of tis constituents is in the form if
serum, plasma, whole blood and protein-free filtrate. If blood is removed from any vessel, it forms clots
or coagulates in a few minutes. The whole mass becomes gelatinous. If left undisturbed, a clear straw-
colored fluid is gradually squeezed out of the whole blood; this is called serum.
Blood plasma can be separated from formed elements an anticoagulant is added immediately after
the blood has been obtained. If it is centrifuged, a fluid that is also clear and straw color is obtained. This
is called plasma.
Since the whole blood is made of formed elements and plasma, it also contains proteins which
must be removed in order not to interfere with colorimetric determination. Such blood sample is treated
with substance to remove protein. The resulting solution is clear filtrate know as protein-free filtrate
(PFF).
Whole blood – formed elements = plasma
Whole blood – (formed elements – clotting factors) = serum
Plasma – protein = protein-free filtrate
Many substances are used to precipitate protein for the blood analyses. The choice of appropriate
method for PFF preparation depends on many factors, such as (1) type of proteinaceous substances to be
precipitated, (2) other constituents affected, (3) possible interference of access precipitant in the filtrate on
the later stages in the procedure, (4) ease of removal of precipitate from protein free filtrate. In general,
1mL of blood, plasma, or serum will yield about 6mL of filtrate in a 1:10 dilution.
Objective
At the end of the activity, the students will be able to
3. prepare a protein-free filtrate
4. differentiate serum from plasma
5. differentiate and identify the different methods in the preparation of protein free filtrate
Materials
9. Blood Extraction set
10. 5mL and 20mL test tubes
11. Test tube rack
12. Serologic pipet (5mL, 1mL)
13. Automatic pipet (5mL, 1mL)
14. Aspirator
15. Centrifuge
16. Applicator stick
Reagents
1. 10% Sodium Tungstate
2. 2/3N Sulfuric Acid
3. 0.3N Barium Hydroxide
4. 5% Zinc Sulfate
5. 5% Trichloroacetic Acid
6. Distilled water
Procedure
1. Prepare the following samples:
Tube 1 – serum
Tube 2 – plasma (3.8% Sodium Citrate)
Tube 3 – whole blood (3.8% Sodium Citrate)
2. Follow the following methods in the preparation of protein – free filtrate
A. Folin Wu Method
1. In three (3) 20mL test tube, measure 4.5mL of 10% Sodium Tungstate and 4.5mL of 2/3N
Sulfuric Acid
2. Add 1mL of the following and mix gently avoiding foam formation:
Tube 1 – serum
Tube 2 – plasma
Tube 3 – whole blood
3. Centrifuge at 2500rpm for 10 minutes
4. Collect the supernatant, put in a test tube label as tube A, B and C respectively
B. Nelson – Somogyi Method

1. Measure 5mL distilled water, 2mL 0.3 N Barium Hydroxide, 2mL 5% Zinc Sulfate in a
20mL test tube and mix
Tube 1 – serum
Tube 2 – plasma
4. Collect the supernatant, put in a test tube label as tube D, E and F respectively
C. Trichloroacetic Acid (TCA)

1. Measure 9mLof 5%TCA each in a 20mL test tube and mix
Tube 1 – serum
Tube 2 – plasma
4. Collect the supernatant, put in a test tube label as tube G, H and I respectively

A. Neutralization
If liquids meet all standards for the sanitary sewer except for pH, campus employees may
neutralize the solution before pouring down the drain. Use proper equipment. Goggles, gloves,
apron, and hood are required. Add neutralizing agent slowly, stirring constantly.
Acidic solutions ( pH <5)

Adjust the pH to 5-9 using a dilute solution (e.g. KOH, NaOH, NaHCO3). Use a pH
meter, indicator solution, or pH paper to determine the pH.
Basic solutions ( pH > 9)
Adjust pH to 5-9 using a dilute solution (e.g. HCl, H2SO4, HNO3 ). Use a pH meter,
indicator solution, or pH paper to determine pH.
Note: For highly concentrated acids, neutralization with a relatively dilute basic solution will take
a very large volume of base and a long time. In this case, consider neutralization using a
concentrated basic solution with plenty of ice for an ice bath, performed slowly and carefully and
with constant stirring. Monitor the temperature of the solution with a suitable thermometer to
ensure that the solution doesn't get too hot. The same is true for neutralizing some concentrated
bases.
B. Safely Sewered
Small amounts of this material may be suitable for sanitary sewer or trash disposal. Non-
hazardous liquid, for the most part, can be poured down the drain. In this case, non-infectious
blood is drained in the sink.
Non-hazardous solid waste can usually be put into the garbage, taking into consideration the
proper segregation of waste. Place the cotton, tissue, syringe barrel and plunger and anything
soiled with contamination of blood and other body fluids in a yellow (infectious) waste container.
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________

Paste your results on the box provided and describe its appearance:
A. Folin Wu Method
Test Tube Color Transparency Filtrate
Tube A
Tube B
Tube C
B. Neslon Somogyi Method

Tube D
Tube E
Tube F
C. Tricholoracetic Acid (TCA)
Tube G
Tube H
Tube I
Post Laboratory Questions

1. Discuss the principle involved in each method of Protein-Free Filtrate Preparation
2. Compare and contrast the serum, plasma and whole blood based on physical appearance.
3. Compare and contrast the different methods of protein-free filtrate preparation.
Laboratory Exercise VII

PLOTTING OF QUALITY CONTROL USING LEVEY – JENNINGS CHART
Introduction
Quality Control (QC) is the process of monitoring laboratory analysis to ensure accuracy of
results. It refers to the technique and procedure that is designed to detect, reduce, and correct deficiencies
in the internal analytical process conducted in the laboratory. This is done prior to the release of any
laboratory results to ensure reliability of each measurement performed on a sample.
QC involves a statistical process which is used to monitor and evaluate the analytical process.
The statistical process generates quality control data that require correct interpretation. QC is usually
presented in a form of graph or chart known as Levey-Jennings chart. The dates of analysis are plotted
along the x-axis and control values are plotted on the y-axis. The mean, 1SD, 2SD and 3SD are also
marked in the y-axis.
Objective
1. understand the principle of Quality Control
2. construct and use Quality Control Chart using Levey – Jennings
3. interpret the chart with application of Westgard rules
4. calculate for the given laboratory data using the basics in statistics
Materials
1. Graphing paper
2. Pencil
3. Ruler
4. Drawing materials
5. Calculator
Procedure
1. Solve for the Mean
2. Compute for the standard deviation and identify the confidence limits up to ± 3 SD
3. Designate at the Y-axis the observed values and dates at the X axis (the ranges of the observe
values follows the computed confidence limits)
4. Mark the mean parallel to the X axis and make it bold to make it visible and identifiable.
5. Locate and label the ranges of the x ± 1 SD, x ± 2 SD, and x ± 3 SD. Mark the following
with green, red and blue lines respectively
6. Plot the values and connect the dots to form a line
7. Interpret the plotted graph according to the Westgard rule
8. Identify the shift, trend and outliers
Formulas:
Mean = Summation of observed units (x)

Number of data (n)
Standard Deviation = √ ∑
❑
❑( x−X )
2
n−1
Confidence Limits:
1. ± 1SD
Upper limit : + 1 SD = x + 1 SD
Lower limit : - 1 SD = x – 1SD
2. ± 2 SD
3. ± 3 SD
Number of days Observed value Number of Days Observed value

1 15.6 16 16.6
2 16.0 17 16.8
3 16.1 18 15.8
4 16.7 19 16.3
5 16.6 20 16.7
6 17.0 21 17.1
7 17.0 22 16.0
8 16.5 23 16.2
9 16.7 24 16.8
10 16.9 25 15.9
11 16.2 26 16.2
12 16.8 27 16.7
13 16.1 28 16.7
14 16.5 29 16.6
15 17.0 30 17.0
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________

Group No: ______________
Complete the table
Number Observed Mean value Difference (x-x) Difference squared (x-

value x)2
1 15.6
2 16.0
3 16.1
4 16.7
5 16.6
6 17.0
7 17.0
8 16.5
9 16.7
10 16.9
11 16.2
12 16.8
13 16.1
14 16.5
15 17.0
16 16.6
17 16.8
18 15.8
19 16.3
20 16.7
21 17.1
22 16.0
23 16.2
24 16.8
25 15.9
26 16.2
27 16.7
28 16.7
29 16.6
30 17.0
Total
Place your graph here
Interpretation
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_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
1. Differentiate the following:

Trends
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
Shift
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
Outliers
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
2. What is a control?
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
3. Enumerate some examples of Pre – Analytical, Analytical and Post-Analytical variables
Pre Analytical Analytical Post Analytical
4. Enumerate the Westgard Rules and its interpretation

Westgard Rules Interpretation
5. Name and describe the other Charts used in describing Quality Control
Laboratory Exercise VIII

SPECTROPHOTOMETER
Introduction
The spectrophotometer uses light to measure the concentration in a solution. When interacting with
the solution, light can be reflected by the tube, transmitted through the solution, or absorbed by the
molecules in the solution. When you see a color of an object, it is because the molecules of that object
have absorbed all the wavelengths of light except the wavelength associated with the color that you see.
Therefore, a green leaf absorbs all colors of light except green which is reflected back to your eye.
Finding the wavelength that absorbs the maximum amount of light (lambda max) can be used to determine
the concentration of a molecule in the solution as there is a direct, linear relationship between the
absorbance and the concentration of the molecules.
Materials
Spectrophotometer
Cuvettes
Colored reagents
Filter paper
Test tubes
Precautions
- Turn on the spectrophotometer 15minutes prior to use to allow it to warm up
- Clean off the test tubes used as cuvettes t remove fingerprints and oils
Procedures
Part A: Discovering the Spectrophotometer
1. Cut a strip of filter paper to fit into a 13x100 mm glass tube
2. Place the tube in the sample holder so that the inside of the fold faces to the right
3. Set the mode to 100% Transmittance and adjust the wavelength to 400nm
4. With the sample holder open, cup your hands around it and look down into the tube
5. Have your partner slowly adjust the wavelength dial and note the color changes that you see
6. Record the colors and the corresponding wavelength that they appear
Part B: Absorption spectrum

A. Prepare the following solutions
Copper (II) sulphate pentahydrate. Different groups can be given an assigned weight of
sample from the following choices: 0.499, 0.936, 1.86, 2.711 and 3.556 mg. Place the
assigned amount into 100mL volumetric flask and fill with distilled water up to the mark.
Shake until the salt is dissolved.
Nickel (II) sulphate hexahydrate. Different groups can be given an assigned weight of sample
from the following choices: 1.262, 2.450, 3.655, 4.813 and 5.944 mg. Place the assigned
amount into 100mL volumetric flask and fill with distilled water up to the mark. Shake until
the salt is dissolved.
B. Do the spectrophotometric reading of these solutions
1. Set and warm up the spectrophotometer for 15-20 minutes.
2. Get two clean and dry cuvettes. Fill one with distilled solution and the other with one
sample of copper salt solution, or with nickel salt solutions after rinsing the cuvette
with solution three times. Wipe the outside of each cuvette with tissue
3. Set the instrument to zero absorbance (100% transmittance)
4. Adjust the wavelength to 400nm for copper (II) sulphate and 350nm for nickel
sulphate
5. Insert the cuvette with distilled water into the cell holder and cover. Set again to 100%
transmittance
6. Read the sample absorbance after inserting the cuvette containing the sample into the
cell holder
7. Repeat steps 5 and 6 for each wavelength used. Take the absorbance readings using
the given wavelength below.
Copper sulphate pentahydrate Nickel sulphate
400 nm 350 nm
500 nm 400 nm
550 nm 450 nm
600 nm 500 nm
650 nm 550 nm
8. After all sample solutions have been measured, turn off the spectrophotometer
9. Using a graphing paper, plot the absorbance versus wavelength for all solutions
containing copper sulphate and separate graph for nickel sulphate solutions
Safely Sewered
Certain heavy metals as listed below with restrictions on concentration and quantity. If both
concentration and quantity amounts are exceeded, liquid waste cannot be sewered:
Metal Concentration
MG/L Pounds/24 hours
Arsenic 0.5 0.2
Cadmium 2.0 0.8
Chromium (total) 10.0 4.0
Copper (total) 5.0 2.0
Cyanide (total) 5.0 2.0
Mercury 0.02 -
Nickel 10.0 4.0
Zinc 15.0 6.3
Acrylonitrile 1.0 -
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________

Complete the table
Wavelength Color UV / Infrared
400nm
450nm
500nm
550nm
600nm
650nm
700nm
Copper Sulfate solution

Wavelength Absorbance of the assigned weights
(nm) Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
400
450
550
600
650
Nickel Sulfate solution

Wavelength Absorbance of the assigned weights
(nm) Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
350
400
450
500
550
Plot the absorbance (y-axis) reading versus wavelength (x-axis).
Copper sulphate solution
Nickel sulphate solution
Laboratory Exercise VIII

GLUCOSE DETERMINATION
Introduction
Carbohydrates are the primary source of energy of the body. Assessment of the state of the
carbohydrate production, metabolism, and excretion primarily involves the measurement of glucose in
body fluids, especially in blood. Early recognition and diagnosis of diabetes mellitus allows better
management and thus prevents complications. The normal blood glucose level is approximately 70-100
mg/dL depending on the assay method variation. Any method used to quantittate glucose in serum or
plasma must be able to measure this material accurately throughout a wide range of values and ensures
specificity. This method used to quantitate glucose can have considerable influence on the apparent
differences between plasma glucose and whole blood. Much of the variation can be attributed to
interfering substance in red cells. Approximately, plasma glucose can be considered 10-15% higher than
corresponding whole blood concentration.
Objective
At the end of the activity, students will be able to:
1. perform glucose determination using O-toluidine method
2. differentiate the methods of glucose determination
3. understand the principle of glucose determination
PRINCIPLE OF THE METHOD

Glucose in the sample originates, by means of coupled reactions described below, a colored complex that
can be measured by spectrophotometry.
Glucose + ½ O2 + H2O Glucose oxidase Gluconated + H2O2
2 H2O2 + 4 – Aminoantipyrine + Phenol peroxidase Quinoneimine + 4H 2O2
COMPOSITION
A. Reagent: Phosphate 100 mmol/L, phenol 5 mmol/L, glucose oxidase > 10 U/mL, peroxidase
1U/mL, 4 – aminoantipyrine 0.4 mmol/L, pH 7.5
A. S. Glucose/Urea/Creatinine standard. Glucose 100md/dL, urea 50mg/dL, creatinine 2 mg/dL(177
µmmol/L). Aqueous primary standard.
STORAGE
Store at 2-8 °C.
Reagent and standard is stable until expiry date shown on the label when stored tightly closed and if
contaminations are prevented during their use.
Indications of deterioration:
- Reagent: presence of particulate material, turbidity, absorbance of the blank over 0.150 at 500 nm
(1 cm cuvette)
- Standard: presence of particulate material, turbidity.
ADDITIONAL EQUIPMENT
- Thermostatic water bath at 37°C
- Analyzer, spectrophotometer or photometer able to read 500 ± 20 nm
SAMPLES
Serum or plasma collected by standard procedures
Serum or plasma must be separated from the red cells promptly to prevent glycolysis. The addition of
sodium fluoride to the blood sample prevent glycolysis.
Cholesterol in the serum or plasma is stable for 5 days at 2-8 °C. Heparin, EDTA, oxalate and fluoride
may be used as anticoagulants
Cerebrospinal fluid collected by standard procedures. Cerebrospinal fluid may be contaminated with
bacteria or other cells and should therefore be analyze with glucose immediately
PROCEDURE
1. Bring reagent to room temperature.

2. Pipette into labelled test tubes (Note 1).
Blank Standard Sample

Cholesterol Standard ---------------- 10 µl -----------------
-
Sample ---------------- ---------------- 10 µl
- -
3. Reagent A 1.0 mL 1.0 mL 1.0 mL Mix
thoroughly and incubate the tubes for 10 minutes at room temperature (16-25 °C) or for 5 minutes
at 37 °C.
4. Measure the absorbance (A) of the standard and sample at 500 nm against the blank. The color is
stable for at least 2 hours .
CALCULATIONS
The cholesterol concentration in the sample is calculated using the following general formula:
A sample
X C standard = C sample
A standard
If the cholesterol standard provided has been used to calibrate (Note 2):
A sample X 100 = mg/dL

A standard X 5.55 = mmol/L
REFERENCE VALUES
Serum and Plasma
Newborn, premature 25 – 80 mg/dL = 1.39 – 4.44 mmol/L
Newborn, term 30 – 90 mg/dL = 1.67 – 5.00 mmol/L
Children, adult 70 – 105 mg/dL = 3.89 – 5.83 mmol/L
Cerebrospinal Fluid
Children 60 – 80 mg/dL = 3.33 mmol/L
Adult 40 – 7- mg/dL = 2.22 – 3.89 mmol/L
SITUATION 2 1 0
1. Correct venipuncture technique
2. Proper preparation of serum, which is not hemolysed.
3. Proper labelling of tubes.
4. Proper measurement of reagents and samples with respective pipettes.
5. Accurate measurement of the volumes of samples and reagents,

executing correct pipetting techniques.
6. Correct setting of temperature of the water bath.
7. Correct observation of incubation period.
8. Correct absorbance reading obtained, observing the correct sequences
of the tubes in the set up.
9. Correct calculation of results.
10. Correct correlation of results obtained that are consistent with colors
developed in the reaction.
11. Correct Preparation of result (form) and submitted on time, vis-à-vis
with checking of assay set up.
12. Proper disposal of lab wastes.
13. Maintenance of area of that is kept clean and orderly.
14. Preparedness in the performance of the experiment.
TOTAL POINTS
Score
Safely Sewered
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________
Paste a picture of the following in the box provided

Results Observation
Blank
Standard
Sample
Calculate for the final value
Interpretation
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

1. Name the different methodologies in glucose determination, give its principle and identify its
advantage and disadvantages
2. Give the sources of error in glucose testing
3. Give and differentiate the different specimen for glucose testing
Laboratory Exercise X
TOTAL PROTEIN DETERMINATION

Protein in the sample reacts with copper (II) ion in alkaline medium forming a colored complex that can
be measured by spectrophotometry
COMPOSITION
A. Reagent. Copper (II) acetate 6 mmol/L, potassium iodide 12 mmol/L, sodium hydroxide 1.15 mol/L
Protein Standard (S). Bovine Albumin. Concentration is given on the label. Concentration value is
traceable to the standard reference material 927 (NIST)
STORAGE
Reagent (A) Store at 15 – 30 degree Celsius
Protein Standard (S) Store at 2 – 8 degree Celsius
Reagents and Standard are stable until the expiry date shown on the label when stored tightly closed and
if contaminations are prevented during their use.
(1 cm cuvette)
SAMPLES
Serum, or heparinized plasma collected by standard procedures.
Stable for 8 days at 2-8 degrees Celsius.
Anticoagulants other than heparin should not be used.
PROCEDURE
1. Pipette into labeled test tubes: (Note 1)

1. Blank Standard Sample
Distilled water 20 µl ---------------- -----------------
-
Protein Standard ---------------- 20 µl -----------------
-
Sample ---------------- ---------------- 20 µl
- -
Reagent A 1.0 mL 1.0 mL 1.0 mL
2. Mix thoroughly and let stand the tubes for 10 minutes at room temperature.
3. Read the absorbance (A) of the standard and the sample at 545 nm against the blank. The color is
stable for 2 hours.
CALCULATIONS
The Creatinine concentration in the sample is calculated using the following general formula:
A sample
A standard
REFERENCE VALUES
Serum and Plasma
Ambulatory 64 – 83 g/L
Recumbent 60 – 78 g/L
Laboratory Exercise XI
ALBUMIN DETERMINATION

Albumin in the sample reacts with bromcresol green in an acid medium, forming a colored complex that
can be measured by spectrophotometry.
COMPOSITION
A. Reagent: Acetate buffer 100 mmol/L, bromcresol green 0.27 mmol/L detergent pH 4.1
S. Albumin standard: Bovine Albumin Concentration is given on the label
STORAGE
Reagent (A) Store at 2-8 °C.
Albumin Standard (S) Store at 2-8 °C.
(1 cm cuvette)
- Analyzer, spectrophotometer or photometer able to read 630 nm.
SAMPLES
Serum or plasma (EDTA, Heparin, Citrate ) collected by standard procedures
PROCEDURE
1. Pipette into labeled test tubes: (Note 1)

Albumin Standard ---------------- 10 µl -----------------
-
Sample ---------------- ---------------- 10 µl
- -
Reagent A 1.0 mL 1.0 mL 1.0 mL
2. Mix thoroughly and let stand the tubes for 1 minute at room temperature.
3. Read the absorbance (A) of the standard and the sample at 630 nm against the blank. The color is
stable for 30 minutes.
CALCULATIONS
The Albumin concentration in the sample is calculated using the following general formula:
A sample
A standard
REFERENCE VALUES
Serum
Newborn, 2 to 4 days 28 – 44 g/L

4 days to 14 years 38 – 54 g/L
Adult 45 – 50 g/L
>60 years 34 – 38 g/L
Laboratory Exercise XII

BLOOD UREA NITROGEN DETERMINATION (BUN)
Introduction
The major substance normally excreted by the kidneys comprises the Non protein nitrogen. Its
major component urea nitrogen which is about 45% of total non protein nitrogen. Other NPN include:
amino acids, uric acid, creatinine, creatine and ammonia. These substances are arranged in the order of
their decreasing quantitative distribution. These substances are arranged in the order of their decreasing
quantitative distribution. Determination of NPN is used to evaluate renal blood flow.
Urea is the end-product of the ornithine-arginine-urea cycle which is synthesized in the liver and
may rise from normal catabolism of endogenous exogenous (dietary proteins). Urea is freely diffusible
across cell membranes and most are excreted by the bacteria in the intestine.
Objective
1. be able to perform BUN testing using colorimetric reaction
2. illustrate other the principle involved in BUN determination
3. identify the clinical significance of BUN
Method
Urease/Salicylate
Principle
Urea in the sample originates by means of the coupled reaction where a coloured complex is measured
spectrophotometrically
Urea + H2O gluconic acid + H2O2
urease
NH4 + Salicylate + NaClO nitroprusside Indophenol
Materials
Test tube (5mL) Distilled water
Spectrophotometer Centrifuge
Automatic Pipettor Disposable tips
Timer Waterbath
Serologic pipet (5mL, 1mL) Aspirator
Blood extraction set
Procedure
1. Bring reagent to room temperature
2. Pipette into labelled tubes
BLANK STANDARD SAMPLE
BUN Standard 10uL
Sample 10uL
Reagent A 1mL 1mL 1mL
3. Mix thoroughly and incubate the tubes for 10 minutes at room temperature (16-25 minutes) or for 5
minutes at 37°C
4. Pipet 1mL of Reagent A on all tubes
5. Mix thoroughly and incubate the tubes for 10 minutes at room temperature (16-25 minutes) or for 5
minutes at 37°C
6. Measure the blank, standard and the sample spectrophotometrically. The color is stable for two
hours.
Concentration of Unknown (mg/dL) = Absorbance of Unknown x Concentration of standard

Absorbance of standard
Reference range: 1.65 – 8.13 mg/dL

Conversion factor: 0.0357 (mg/dL to mmol/dL)
Safely Sewered
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________

Results Observation
Blank
Standard
Sample
Interpretation
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

1. Name the different methodologies in urea determination, give its principle and identify its
SITUATION 2 1 0

TOTAL POINTS
Score
Laboratory Exercise XIII
BLOOD URIC ACID DETERMINATION (BUA)
Introduction
Uric Acid is the final product of purine catabolism in humans. Most of the uric acid in the body
dissolves in blood and travel to the kidneys. It is removed from the body in urine. A small amount of uric
acid appears in stool. If too much uric acid is being produced, its level in the urine will increase. High
blood levels of uric acid in the body can lead to a condition known as gout, which is characterized by the
disposition of solid crystals of uric in the joints. Determination of blood uric acid level is helpful tool in
the diagnosis of gout.
Objective
1. be able to perform BUA testing using colorimetric reaction
2. describe other the principle involved in BUA determination
3. understand the metabolism of BUA
4. determine the clinical significance of BUA
Method
Uricase/Peroxidase
Principle
Uric Acid in the sample originates by means of the coupled reaction where a coloured complex is
measured spectrophotometrically
Uric Acid + O2 + H2O uricase Alantoin + CO2 + H2O2
2 H2O2 + 4-Aminoantipyrine + DCFS peroxidase Quinoneimine + H2O2
Materials
Test tube (5mL) Distilled water
Spectrophotometer Centrifuge
Automatic Pipettor Disposable tips
Timer Waterbath
Serologic pipet (5mL, 1mL) Aspirator
Blood extraction set
Reagents
1. 2/3 N Sulfuric Acid
2. 10% Sodium Tungstate
3. 14% Sodium Carbonate
4. Phosphotungstic acid
Procedure
1. Prepare a Protein Free Filtrate serum sample using Folin Wu method
2. Prepare a serum sample
3. Pipette into labelled tubes
Tube No (1) BLANK (2) STANDARD SAMPLE
Distilled Water 3mL
BUA Standard 1mL
Serum 1mL (tube 3)
PFF 3mL (tube 4)
14% Sodium Carbonate 1mL 1mL 1mL
4. Mix thoroughly and incubate the tubes for 15 minutes at room temperature
5. Measure the blank, standard and the sample spectrophotometrically at 710nm. The color is stable
for 30 minutes.
Concentration of Unknown (mg/dL) = Absorbance of Unknown x Concentration of standard

Absorbance of standard
Reference range:
Male: 3.4 – 7.0 mg/dL
Female: 2.4 – 5.7mg/dL
Conversion factor: 59.6 (mg/dL to mmol/L)
Safely Sewered
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________

Results Observation
Tube 1
Tube 2
Tube 3
Tube 4
Interpretation
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

2. Name the different methodologies in uric acid determination, give its principle and identify its
SITUATION 2 1 0

TOTAL POINTS
Score
Laboratory Exercise XIV

CREATININE DETERMINATION
Introduction
Creatinine is the final product of creatinine phosphate in muscles. As creatinine is produced, it is
filtered through the kidneys and excreted in urine. Creatinine is made at a steady rate and is not affected
by diet or by normal physical activities. If the kidneys are damaged and cannot filter creatinine, its
amount in the urine is decreased and its level in the blood is increased. Blood creatinine determination
can be used to test for kidney function. Compared to blood urea nitrogen, creatinine determination is
more dependable diagnostic tool since creatinine is not dependent with protein dietary contents, on which
urea levels are dependent. Thus, creatinine determination offers a more reliable screening rest for renal
pathology and index of overall renal function.
The Jaffe reaction is used as a colorimetric method in clinical chemistry to determine creatinine
levels in blood and urine. In this reaction, creatinine reacts with alkaline picrate to produce a reddish-
orange colored solution, the intensity of which at 490nm is directly proportional to the creatinine
concentration.
Objective
1. be able to perform Creatinine test using colorimetric reaction
2. learn other the principle involved in Creatinine determination
3. describe the metabolism of creatinine
4. describe the method of creattinine clearance test

Creatinine is a sample that reacts with picrate in an alkaline medium forming a colored complex (Jaffe
method). The complex formation rate is measured in a short period to avoid interferences. Serum and
plasma samples contain proteins that react in a non specific way; nevertheless, the results can be corrected
subtracting a fixed value. The use of this correction is known as the Jaffe method compensated.
COMPOSITION
B. Reagent: Sodium hydroxide 0.4mol/L detergent
C. Reagent: Picric acid 25mmol/L
D. Glucose/Urea/Creatinine standard. Glucose 100md/dL, urea 50mg/dL, creatinine 2 mg/dL(177
ummol/L). Aqueous primary standard.
STORAGE
Store at 15 – 30 degree Celsius
Reagents and Standard are stable until the expiry date shown on the label when stored tightly closed and
if contaminations are prevented during their use.
(1 cm cuvette)
SAMPLES
Serum, plasma or urine collected by standard procedures. Dilute fresh urine 1/50 with distilled water
before measurement. Heparin, EDTA, oxalate and fluoride may be used as anticoagulants
Creatinine in samples is stable for 24 hours at 2-8 degrees Celsius.
PROCEDURE
1. Bring the working reagent and photometer to 37°C
2. Pipette into labeled tubes: (Note1)
Working reagent 1.0 mL
Standard (S) or Sample 0.1 mL
3. Mix and insert cuvette in the photometer. Start stop watch

4. Record absorbance at 500 nm after 30 seconds (A1) and after 90 seconds (A2)
CALCULATIONS
The Creatinine concentration in the sample is calculated using the following general formula:
(A2 – A1) Sample X C standard x Sample dilution factor
(A2 –
-Corrective factor 4,5,6 = C sample
A1)Standard
If the Creatinine standard provided has been used to calibrate (Note 2):
Serum and Plasma Urine

Jaffé Non -compensated Jaffé Compensated
X 2 = md/dL X2 – 0.37 = mg/dL X 100 =mg/dL
(A2 – A1) Sample
X 117 = µmol/L X 177 – 33 = X 8840= µmol/L
µmol/L
(A2 – A1)Standard
REFERENCE VALUES
Serum and Plasma
Method Jaffé Non -compensated Jaffé Compensated
Men 0.9 – 1.3 mg/dL = 80 – 115 µmol/L 0.7 – 1.2 mg/dL = 62 – 106 µmol/L
Women 0.6 – 1.1 mg/ dL = 53 – 97 µmol/L 0.5 -0.9 mg/dL = 44 – 80 µmol/L
Urine:
Men: 14 – 26 mg/kg/24hr = 124 – 230 µmol/kg/24hr
Women: 11 – 20 mg/kg/24hr = 97 – 177 µmol/kg/24hr

Safely Sewered
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________

Results Observation
Tube 1
Tube 2
Tube 3
Tube 4
Tube 5
Interpretation
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

1. Name the different methodologies in creatinine determination, give its principle and identify its
SITUATION 2 1 0

TOTAL POINTS
Score
Laboratory Exercise XV
LIPID DETERMINATION
Introduction
The most commonly employed lipid analyses are triglycerides and cholesterol determination. An
understanding of the concentration of these substances in serum and plasma is used as an index in the
diagnosis and management of certain diseases and disorders associated with lipid metabolism.
Triglyceride determination is carried out by enzymatic method. Here, lipoprotein lipase is
required to produce glycerine and fatty acid. Glycerine will then participate in the series of coupled
reactions to yield the desired colored complex which is proportional to the concentration of unknown
triglycerides.
Modified Ferro-Ham method is the most commonly used method got cholesterol determination.
This method employs the Liebermann-Burchardt reaction which gives a green color. This color is directly
developed without prior extraction of the lipids.
Objective
At the end of the activity, students will be able to:
1. perform triglyceride and cholesterol determination using colorimetric reactions
2. differentiate the methods of lipid determination
3. understand the principle of triglyceride and cholesterol determination
TRIGLYCERIDES

Triglyceride in the sample originates, by means of the coupled reactions described below, a colored
complex that can be measured by spectrophotometry.
Triglycerides + H2O lipase Glycerol + Fatty Acids
Glycerol + ATP Glycerol kinase Glycerol – 3 – P + ADP
Glycerol – 3 – P + O2 G-3-P-oxidase Dihydroxiacetone – P + H2O2
2 H2O2 + 4 – Aminoantipyrine + 4 – Clorophenol peroxidase Quinoneimine + 4H2O2
COMPOSITION
A. Reagent: Pipes 45 mmol/L, magnesium chloride 5 mmol/L, 4 chlorophenol 6 mmol/L, lipase
>100 U/mL, glycerol kinase >1.5 U/mL, glycerol-3-phosphate oxidase >4 U/mL, peroxidase >0.8
U/mL, 4-aminoantipyrine 0.75 mmol/L, ATP 0.9 mmol/L, ph 7.0
B. Triglyceride standard: Glycerol equivalent to 200 mg/dL (2.26 mmol/L) triolein. Aqueous
primary standard.
STORAGE
Store at 2-8 °C.
Reagent and A sample standard is
stable until A standard expiry date
shown on the label when stored tightly closed and if contaminations are prevented during their use.
(1 cm cuvette)
SAMPLES
Triglyceride in the serum or plasma is stable for 5 days at 2-8 °C. Heparin, EDTA, oxalate and fluoride
may be used as anticoagulants.
PROCEDURE
1. Bring the reagent to room temperature
2. Pipette into labeled tubes: (Note1)
Triglycerides standard ----- 10μL ----
Sample ----- ------ 10μL
Reagent A 1.0mL 1.0mL 1.0mL
3. Mix thoroughly and incubate the tubes for 15 minutes at room temperature (16-25 °C) or for 5
minutes at 37 °C.
stable for at least 2 hours.
CALCULATIONS
The triglyceride concentration in the sample is calculated using the following general formula:
If the triglycerides standard provided has been used to calibrate (Note 2):
A sample X 200 = mg/dL Triglycerides

A standard X 2.26 = mmol/L Triglycerides
REFERENCE VALUES
The following uniform cutoff points have been established by the US National Institutes of Health and
have also been adopted in many countries for the evaluation of the risk.
Up to 150 mg/dL = 1.7 mmol/L Normal

150-199 mg/dL = 1.70 – 2.25 mmol/L Borderline High
200-499 mg/dL = 2.26 – 5.64 mmol/L High
>500 mg/dL = >5.65 mmol/L Very High
CHOLESTEROL

Free and esterified cholesterol originates, by means of coupled reactions described below, a colored
complex that can be measured by spectrophotometry.
Cholesterol
esterase
Cholesterol ester + H2O Cholesterol + Fatty Acid
Cholesterol oxidase
Cholesterol + ½ O2 + H2O Cholestenone + H2O2
2 H2O2 + 4 – Aminoantipyrine + DCF peroxidase Quinoneimine + 4H 2O2
COMPOSITION
A. Reagent. Pipes 35 mmol/L, cholesterol esterase >0.2 U/mL, cholesterol oxidase > o.1 U/mL,
peroxidase > 0.2 U/mL, 4 – aminoantipyrine 0.5 mmol/L, sodium cholate 0.5 mmol/L,
dicholorophenolsulfonate 4 mmol/mL, pH 7.0
S. Cholesterol standard: Cholesterol 200 mg/mL (5.18 mmol/L). Aqueous primary standard.
STORAGE
Store at 2-8 °C.
(1 cm cuvette)
SAMPLES
Cholesterol in the serum or plasma is stable for 7 days at 2-8 °C. Heparin, EDTA, oxalate and fluoride
may be used as anticoagulants.
PROCEDURE
1. Bring reagent to room temperature.

2. Pipette into labelled test tubes (Note 1).
1. Blank Standard Sample

Cholesterol Standard ---------------- 10 µl -----------------
-
Sample ---------------- ---------------- 10 µl
- -
3. Reagent A 1.0 mL 1.0 mL 1.0 mL Mix
thoroughly and incubate the tubes for 10 minutes at room temperature (16-25 °C) or for 5 minutes
at 37 °C.
stable for at least 2 hours .
CALCULATIONS
The cholesterol concentration in the sample is calculated using the following general formula:
A sample
A standard
If the cholesterol standard provided has been used to calibrate (Note 2):
A sample X 200 = mg/dL HDL cholesterol

A standard X 5.18 = mmol/L HDL cholesterol
REFERENCE VALUES
Cholesterol concentrations vary considerably with age and sex. The following cut – off point has been
recommended for identifying individuals at high risk of coronary artery disease.
Up to 200 mg/dL = 5.2 mmol/L Desirable
200 – 239 mg/dL = 5.2 – 6.21 mmol/L Boarderline

High
>240 mg/dL = 6.24 mmol/L HIgh
LABORATORY REPORT
Name: ___________________________________________________ Date: ______________
Group No: ______________
A. Triglycerides
Results Observation
Blank
Standard
Sample
B. Cholesterol
Results Observation
Blank
Standard
Sample
Interpretation
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

1. Name the different methodologies in Triglycerides and Cholesterol determination, give its
principle and identify its advantage and disadvantages
2. Give the sources of error in triglyceride and cholesterol determination
SITUATION 2 1 0

TOTAL POINTS
Score

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Laboratory Exercise Title Scores

1 Laboratory Safety and Regulations

Preparation of Solutions and Standardization of

3A Measuring small volumes using serologic pipets

3B Measuring small volumes using Automated Pipets

5 Preparation of Blood Samples

6 Preparation of Protein Free Filtrate

Plotting of Quality Control Chart Using Levey-

10 Total Protein Determination

12 Blood Urea Nitrogen Determination

13 Blood Uric Acid Determination

15 Lipid Determination (Triglycerides and Cholesterol)

The grading of laboratory reports / activities shall include:

Tubes Red Green Blue Yellow Total volume Actual % error

% error = expected volume – actual volume x 100

Standard Operating Procedure for Pipetting

Student Performance Checklist

Evaluation of Pipetting Techniques

Pipet the following amount s in the designated assay plate cells

Student Performance Checklist

Evaluation of Pipetting Techniques

Laboratory Waste Disposal

Name: ___________________________________________________ Date: ______________

Results and Observation

3. What are the factors affecting pipetting performance

Laboratory Exercise III

Student Performance Checklist

Laboratory Waste Disposal

Acidic solutions ( pH <5)

Results and Observations

Preparation of 0.1N HCl

Show the computation for the standardization of NaOH

3. How many mL of 8N H2SO4 will neutralize 45mL of 6N KOH?

4. Solve the equivalent weight of H2CO3?

Student Performance Checklist

Evaluation of Tourniquet Application and Vein Selection Competency

Student Performance Checklist

Evaluation of Venipunture using an Evacuated tube Competency

Student Performance Checklist

Evaluation of Venipuncture Using a Syringe Competency

Laboratory Waste Disposal

Name: ___________________________________________________ Date: ______________

Complete the table

Table 1. Order of Draw for Syringe Method

Table 2. Order of Draw for Evacuated Tube Method

1. collection of CBC from a 35-year old woman

Student Performance Checklist

Evaluation of Equipment Selection and Assembly Competency

2 = Satisfactorily Performed 1 = Needs Improvement 0 = Incorrect/Did not perform

B. Nelson – Somogyi Method

C. Trichloroacetic Acid (TCA)

Laboratory Waste Disposal

Acidic solutions ( pH <5)

Results and Observation

B. Neslon Somogyi Method

Test Tube Color Transparency Filtrate

Name: _______________________________________ Section: _

Name: _____________________________________ Date:

Name: _____________________________________ Date:

Name: _____________________________________ Date: