MEDICAL DEGREE MD 3101 LOG BOOK

METABOLISM AND NUTRITION (BIOCHEMISTRY)

SEPTEMBER 2015 INTAKE

No

Page No.
Practical Topic

Optimal enzyme activity

Determination of presence of sugar, ketones

bodies and protein in urine

Urinalysis: Macroscopic, microscopic and urine

14

dipstick test
Estimation of total serum proteins and serum

21

albumin
Venipuncture, intravenous fluid infusion
procedure and blood glucose test (glucometer)

Year 1| Biochemistry | Practical Manual | 1

28

BIOCHEMISTRY LABORATORY SAFETY

Students working in a biochemistry laboratory must always be aware that the
chemicals used are potentially toxic, irritating and flammable. Students who come to
the laboratory session must have a complete understanding of the laboratory
procedures to carry out and be familiar with both the physical and chemical
properties of chemicals and reagents to be used. Become familiar with the location
and the use of standard safety features in the laboratory. The laboratory is equipped
with fire extinguishers, eye washes, safety showers, fume hoods and first-aid kits.
Any question regarding the use of these facilities should be addressed to your
instructor.

Safety rules in general

1. Eating, drinking and smoking in the laboratory are strictly prohibited.
2. Proper attire must be worn at all times.
Wear lab coat, protective glove and protective shoes that can protect your feet
when handling chemicals. Tie and pin long hair, scarves and headwear out of
the way to avoid contact with flames or chemicals.
3. Do not taste, smell, or touch any chemicals/reagents. NEVER put your nose
over the bottle! Avoid looking into the mouth of any reaction vessel in which a
reaction is in progress.
4. While heating a solution one should make sure not to overheat it; therefore,
vigorous mixing of the solution by shaking or stirring is required. The mouth of
the glassware containing the solution to be heated should never be pointed
toward anyone.
5.

Volatile liquids and solids that are toxic or irritating should be handled under
fume hoods.

6. Report ALL injuries, allergies and/or medical problems to the lab instructor.
7.

Before leaving the laboratory, electrical equipment and gas burner should be
turned off.

I, _______________________________________ , have read carefully and

understand the guidelines listed in BIOCHEMISTRY LABORATORY
SAFETY contained on this sheet. I agree to act responsibly and safely
in the laboratory at all times.
Year 1| Biochemistry | Practical Manual | 2
Students signature: _____________________

Date

PRACTICAL 1: OPTIMAL ENZYME ACTIVITY

Introduction
Salivary amylase is a digestive enzyme secreted by the salivary glands.
It is responsible for starting the breakdown of the insoluble
polysaccharide starch into soluble dextrins (oligosaccharides),
maltose (disaccharide) and glucose (monosaccharide) in the mouth so
that starch can be absorbed.
Complete digestion of starch occurs in the small intestine by the action
of pancreatic amylase.

Salivary amylase
Starch

dextrin + maltose + glucose

Objective:
1. To determine the effect of different factors (such as temperature and pH
on enzyme activity)
2. To differentiate the common precipitation and color-changing in enzyme
experiment.
Materials & Reagents:
Materials
Test tubes
Beaker
Ice
Water bath ( 37C and 70C)
Bunsen burner

Procedure:
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Reagents
Distilled water
0.5% sodium chloride (NaCl)
2% starch
Benedict reagent
Alkaline solution
Acidic solution
Neutral solution (normal saline)

Preparing enzyme solution

I. In a beaker, collect about 1 ml of your own saliva
II.
Add 9 ml of distilled water and 60 ml of 0.5% sodium chloride solution
III. gently mix.
IV.
Use this enzyme solution to carry out the following four experiments:
(a) Effect of temperature on the activity of salivary amylase:
Principle:
Salivary amylase is protein in nature; the optimum temperature for its activity
is 37C. In extreme temperatures (cold and heat), the activity of the enzyme
is inhibited (high temperature causes denaturation of the protein structure of
the enzyme).
Procedure:
1. Get 3 test tubes and number them 1, 2 & 3.
2. Put 2 ml of the previously prepared enzyme solution in each of the three
test tubes.
3. To each test tube, add 2 ml of 2% starch solution and mix well.
4. Immediately put:
test tube 1 on ice,
test tube 2 in a 37C water bath
test tube 3 in a 70C water bath
*each for 20 minutes
5. Remove each tube to room temperature
6. Carry out Benedicts test to check for the presence of the starch
hydrolysis product maltose as follows:
add 2 ml of Benedicts reagent to the enzyme-starch mixture in each
tube
put in a boiling water bath for 5 minutes
7. After 5mins, place them in a test tube rack.
8. Wait for few minutes or until the colour change is complete
9. Observe the colour changes. Record all results in the data table

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(b) Effect of pH on the activity of salivary amylase:

Principle:
The optimum pH for the activity of salivary amylase is the neutral pH (about
7). In the acidic pH of the stomach, the activity of salivary amylase stops.
Pancreatic amylase, on the other hand, has an optimum activity in alkaline
medium.
Procedure:
1. Get 3 test tubes and number them 1, 2 & 3.
2. Put 2 ml of the previously prepared enzyme solution in each of the three
test tubes.
3. To each test tube, add 2 ml of 2% starch solution and mix well.
4. For:
Test tube 1, add 2 ml
Test tube 2, add 2 ml
Test tube 3, add 2 ml
*Put the 3 test tubes in

of acidic solution
neutral solution
alkaline solution
a 37C water bath for 10 minutes.

5. Remove each tube to room temperature

6. Carry out Benedicts test to check for the presence of the starch
hydrolysis product maltose as follows:
-add 2 ml of Benedicts reagent to the content of each tube
-put in a boiling water bath for 5 minutes;
7. After 5mins, place them in a test tube rack
8. Wait for few minutes or until the colour change is complete
9. Observe the colour changes. Record all results in the data table

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Results/Observation:
(a)

Effect of temperatures on amylase activity

Temperature

Amylase activity (colour changes)

0C

37C

70C

(b)

Effect of pH on amylase activity

pH

Amylase activity (colour changes)

Acidic
Neutral
Alkaline

Discussion
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a) Effect of temperature

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b) Effect of pH

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Conclusion

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PRACTICAL 2: DETERMINATION OF PRESENCE OF SUGAR,

KETONES
AND PROTEIN IN URINE
OBJECTIVES

To learn how to analyse glucose, ketone and protein of unknown

concentration urine sample
Student will become familiar with common precipitating and color-changing

PRINCIPLE
Benedicts reagent is used as a test for the presence of reducing sugar. This
includes all monosaccharides and the disaccharides, lactose and maltose. Even
more generally, Benedicts test will detect the presence of aldehydes (expect
aromatic ones), and alpha-hydroxy-ketones, including those that accur in certain
ketones. The three main ketone bodies are acetone, acetoacetic acid (diacetic acid)
and beta-hydroxybutyric acid.
INTRODUCTION
To presence of chemical species within urine, with their respective concentrations,
has routinely been used as a diagnostic tool for both illness and death. Nurses and
doctors commonly request a urine or blood sample in an attempt to diagnose health
condition, while toxicologist regularly analyze blood and urine to aid medical
examiners in determination of cause of death.
High levels of proteins, glucose and ketones in urine usually occur within six
potentially fatal medical conditions . Sometimes, elevated levels for pregnancy also
shown. Protein will be high urine for certain cases such as heavy metal poisoning,
kidney failure, diabetes-related and heart failure. While ketones concentration will
be elevated in dehydration cases, starvation and diabetes related. Glucose level in
urine will be increase in kidney failure, diabetes-related and pregnancy-related.
MATERIALS

Bunsen burner
Test tube
Test tube racks
Graduated cylinder
Urine glucose sample
Urine protein sample

Urine ketone sample

REAGENT
Benedict solution
Sodium nitroprusside
Concentrated ammonium sulfate
PROCEDURE
Part A
Obtain test tubes and a test tube rack. Using a graduated cylinder measure out 1ml
of water into a test tube. Mark the water level with a wax pencil or a marker. If all
your test tubes are the same, used the marked tube as a reference and mark all the
others. If the test tubes are different you will need to calibrate each tube separately.
1. Obtain standard simulated urine samples of protein, glucose, and ketones.
Label your samples if this has not already been done.
(a) TEST FOR PROTEINS:
2. Dispense approximately 1-2ml of standard simulated urine protein and 1-2ml
of blank simulated urine sample into two separates tubes.
3. Prepare and light your Bunsen burner. Hold the urine protein test tube over
the flame of the Bunsen burner. Follow al precautions when heating a test
tube in a open flame. Remove the test tube from the flame just as it begin to
boil. The precipitate formed in this experiment appears as soapy bubbles or
froth on the inside of the test tube.
4. Record your observations.
5. Turn off your Bunsen burner . Dispose of your samples in an appropriate
waste container and rinse your test tubes clean.
(b)TEST FOR GLUCOSE (BENEDICTS TEST)
6. Set up apparatus similar into part A, step 11; however, the water will need to
begin boiling for this procedure.
7. Add 5-10 drops of standard simulated urine glucose sample and 5-10 drops of
blank simulated urine into two separate tubes. Additionally, using graduated
disposable pipette, add 2ml of Benedict solution to each test tube.
8. After the water prepared in step 6 has begun to boil, place the test tubes into
the boiling water and allow it to heat for minutes. Remove from heat and
allow it to cool.
9. Record your observation.
10.Turn off your Bunsen burner . Dispose of your samples in an appropriate
waste container and rinse your test tubes clean.

(c) TEST FOR KETONES (ROTHERAS TEST)

11.Dispense approximately 1-2ml of standard simulated urine ketone sample
and 1-2ml of blank simulated urine sample into two separates tubes.
Additionally, add 1ml of Sodium nitropusside solution to each.
12.In a fume hood, slowly add approximately 1ml of concentrated ammonium
hydroxide to each test tube, so as to form a separate layer on top of the
solutions.
13.After 1-2 minutes, record your observation focusing on the interface between
the two layers.
14.Dispose of your samples in an appropriate waste container and rinse your
test tubes clean.

EXPECTED RESULT
(a) Urine protein

(b) Urine glucose

(BENEDICTS TEST)

(c) Urine ketones

(ROTHERAS TEST)

Standard sample
Foamy formation

Blank sample
Clear

Red or green or yellow

precipitate obtained

Clear

Purplish layer form

No colour form

RESULT

DISCUSSION
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CONCLUSION
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PRACTICAL 3: URINALYSIS
(MACROSCOPIC, MICROSCOPIC & URINE DIPSTICK TEST)
Experiment 3(a): Macroscopic analysis
Materials:
1. Urine container
2. Gloves
Procedure:
1. Collect fresh urine specimen into a clean and dry container.
2. Observe and compare the colour and odour between normal and
abnormal urine specimen.
Urine Colour/Appearance
Type
Clear, transparent, pale yellow
Colourless

Milky
Orange

Red

Greenish
Dark brown, brown red, yellow

Brown, brown black, black

Indication
- normal
- hydrated urine.
- diabetes mellitus/insipidus.
- nervousness.
- diuretic.
- alcohol intake.
- purulent genito-urinary tract disease.
- chyluria.
- concentrated urine.
- urobilinogenuria.
- fever.
- excessive sweating.
- beet root ingestion.
- haematuria.
- haemoglobinuria.
- medication (pyridium).
-

jaundice.
phenol poisoning.
very concentrated urine.
acute febrile diseases.
bilirubinuria.
haemorrhage in urinary tract.
haemoglobinuria.
porphyria.
bile pigment

- food and drugs (chloroquine and methyldopa).

Urine Odour
Type
Pungent
Ammonical

Indication
- Normal
- concentrated -dehydrated.
- bacterial action.

Foul

- infection-causing bacteria.

Sweet

- uncontrolled diabetes or a metabolic disorder.

Musty

- liver disease or metabolic disorder.

Fruity

- ketosis.

Results/Observation:
No.
Container 1

Colour

Odour

Container 2

Discussion:
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Conclusion:
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Experiment 3(b): Microscopic analysis

Objective:
i)
To screen the presence of red blood cells, casts, crystals and
bacteria in urine
Principle:
In health, the urine contains small numbers of cells and other formed
elements from the whole length of the genitourinary tract; casts and
epithelial cells from the nephron; epithelial cells from the pelvis, ureters,
bladder and urethra; mucous threads and spermatozoa from the prostate. A
few erythroctyes and leukocytes apparently reach the urine from any part of
the urinary tract. In renal parenchymal disease, the urine usually contains
increased numbers of cells and casts discharged from an organ, which is
otherwise accessible only by biopsy or an operation. The urinary sediment
provides information useful both for diagnosis and prognosis. The presence
of red blood cells, pathological crystals and bacteria as an indicative of
haematuria/ proteinuria/UTI.
Material:
Microscope
Glass slide
Cover slip
Disposable pasteur pipetters

Tissue/paper tissue

Procedure:
i. Using a new disposable pipetter for each sample, pipette out 1.5 to 2
ml into
the clean microscope well unit.
ii.
Centrifuge tubes for five (5) minutes at 400g.
iii.
Remove the supernatant, leaving approximately 1 mL of concentrated
urine
sediment. Mix the sediment well to re-suspend.
iv.
Place a drop of the re-suspended urine sediment onto a glass slide.
v. Place a coverslip over the urine sediment suspension.
vi.
Adjust the microscope condenser down and close the diaphragm
vii.
Hyaline casts and other partially transparent solid elements may be
obscured by excessively bright light.
viii.
Continue scanning using high power objective for crystals , bacteria,
epithelial
cell, mucus, and other formed elements. Count and record the
elements shape

Microscopic Observation

Result Interpretation
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Experiment 3(c): Urine Dipstick Test

Objective:
i)
To screen urine components using dipstick method which provides
results for specific gravity, pH, protein, glucose, and blood in the
urine
ii)
To obtain information of metabolic state, nutritional status, kidney
function and acid-base balance.
Principle:
Urinalysis test strips are easy to use reagent strips for the detection of key
diagnostic chemical markers in human urine. The reagent pads react with the
sample urine to provide a standardized visible color reaction within 30
seconds to one minute depending on the specific panel screen. The color is
then visually compared to the included color chart to determine the level of
each chemical factor. Test results may provide useful information regarding
carbohydrate (sugar) metabolism (diabetes), kidney function, acid-base

balance, bacteriuria, occult blood, high leukocytes (infection) and other

conditions of overall health.
Materials:
1. Urine dipstick strip
2. Urine container
3. Gloves
4. Tissue paper or absorbent paper
Procedure:
3. Collect fresh urine specimen into a clean and dry container.
4. Remove one strip from the bottle and replace cap immediately. Do not
touch the test pads of the strip.
5. Briefly (no longer than one second) immerse all strip/reagent areas into
the urine.
6. Wipe off excess urine on the rim of the container or using the edge of
absorbent paper.
7. Compare the reagent areas to the corresponding color chart on the
bottle label at the time specified. *Hold strip in vertical position.
8. Hold the strip close to the color blocks and match carefully. Proper
reading times are critical for optimal results.

9. Avoid laying the strip directly on the color chart.

From time dipped, after:
30 seconds : glucose
60 seconds : protein, nitrite
2 minutes : leucocytes
10.

Record results in table.

Dipstick Test Result

Dipstick
Paramete
r

Observed result:
Test pad colour
or reading

Negati
ve/
Positiv
e

Indication or Discussion

PRACTICAL 4: ESTIMATION OF TOTAL SERUM PROTEINS

AND SERUM ALBUMIN
OBJECTIVE:

i.

To estimate the total serum protein and total serum albumin

using photometric method (spectrophotometer).

PRINCIPLE:
Principle of Lowry test is based on reaction between peptide nitrogen with the
Copper (II) ions under alkaline condition with the oxidation of aromatic protein
residues. Reduction of Folin- Ciocalteay phosphomolyb diphostungstic acid to
heteropolymolybdenum blue by copper catalysed oxidation of acids.

INTRODUCTION
The protein molecule has a large complex structure, each molecule is made up of
smaller units called amino acids. Amino acids are the building blocks of the protein
molecule. Amino acids are relatively small molecule. They are water-soluble and
consequently can easily diffuse through the walls of the intestine. There are 20
common amino acids. However there are eight amino acids which the body cannot
manufacture successfully. The body needs these hence they are known as the
essential amino acids. They are obtained by eating foods that have these amino
acids. The eight essential amino acids are valine, lysine, leucin, isoleucine,
methionine, phenylalanine, tryotophan, and threonine.
Lowry protein assay is a biochemical assay for determining the total level of protein
in a solution. This method is highly sensitive to low concentration of protein. The
total protein concentration is exhibited by color change of the sample solution in
proportion to protein concentration, which can then be measured using colorimetric
techniques. This method is sensitive to pH changes, therefore it need to be maintain

at 10- 10.5. unlike alkaline conditions the divalent copper ion forms a complex with
peptide bonds in which it is reduced to a monovalent ion. Monovalent copper ion
and the radical groups of tyrosine, tryptophan, and cysteine react with folin reagent
to produce an unstable product that becomes reduced to molybdenum/tungsten
blue.
REAGENT & EQUIPMENTS

Protein kit (Biuret reagent and protein standard)

Albumin kit (BCG reagent and albumin standard)
Spectrophotometer
Cuvette
Micropipette

PRACTICAL 4(A): TOTAL SERUM PROTEIN (Biuret Method)

Principle:
Biuret method has been used to measure total protein in serum. The
formation of a Cuprum-protein complex requires two peptide bonds and
produces colored product which is measured by absorption spectroscopy at
540 560 nm.
pH

Protein (sample/standard) + cuprum

12
(coloured product)

cuprum-protein complex

2 main components that measured in total protein are:

I.
II.

Albumin
Globulin

Low protein is primarily caused by:

glomerular nephritis
nephritic syndrome
protein malabsorption and digestion
cirrhosis of liver

High concentration of protein mainly due :

Multiple myeloma
bone marrow disorder
amyloidosis
chronic inflammatory condition
HIV/AIDS

PROCEDURE
Prepare
I.
II.
III.

3 cuvettes:
Blank
Sample A
Sample B

Pipette
into Microlitre (l)
cuvette
Reagent
1000 l
Sample/ standard 20 l
Mix and incubate for 5 minutes and read the absorbance (using
spectrophotometer) of all cuvettes at 560 nm against reagent blank

Wave length : 540nm

Incubation: 5 minutes
Standard concentration : 4 g/dl
Temperature: 37C

CALCULATION
Absorbance of sample
(g/dl)
Absorbance of standards
EXPECTED VALUE:

Concentration of standard = total protein

= 6.2 to 8.5 g/dl

CALCULATION/RESULT

DISCUSSION
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CONCLUSION

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PRACTICAL 4(B): TOTAL SERUM ALBUMIN

Principle
Serum albumin binds selectively to the dye bromocresol green (BCG) at pH
4.2. The increase in absorbance of resulting albumin-dye complex that read
at 630nm proportional to the albumin concentration.

pH
Albumin (sample/standard) + BCG
(coloured product)
Brij-35

BCG-albumin complex

Albumin will be measured together with TP, aspartate transferase (AST) and
alanine transferase (ALT) to see the liver function (LFT) and usually measured
with creatinine and Blood Urea Nitrogen (BUN) to check Renal Profile (RP).
Low albumin concentration is primarily caused by :
glomerular nephritis
nephritic syndrome
protein malabsorption and digestion
cirrhosis of liver
High concentration of albumin mainly due to :
dehydration
hepatic carcinoma
hypoadrenocorticism

Procedure
Prepare 3 cuvettes:
IV.
V.
VI.

Blank
Sample A
Sample B

Pipette into cuvette

Reagent
Sample/ standard
Mix and incubate for 5
spectrophotometer) of all

Microlitre (l)
1000 l
10 l
minutes and read the absorbance (using
cuvettes at 630 nm against reagent blank

Wave length : 630nm

Incubation: 5 minutes
Standard concentration : 4 g/dl
Temperature: 37C

CALCULATION
Absorbance of sample
(g/dl)
Absorbance of standards
EXPECTED VALUE:
= 3.5 to 5.5 g/dl

CALCULATION/ RESULT

Concentration of standard = albumin

DISCUSSION
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CONCLUSION
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PRACTICAL 5: VENIPUNCTURE, INTRAVENOUS FLUID

INFUSION PROCEDURE
AND BLOOD GLUCOSE TEST (GLUCOMETER)
PRACTICAL 5(a): VENIPUNCTURE PROCEDURE ON
DUMMY/MANNEQUIN
Objective:

Identify and describe equipment used for routine venipuncture.

Assess and select suitable sites used for venipuncture as well as

locations to avoid.

Describe and utilize the required steps to perform routine

venipuncture.

Understand the factors/errors that affecting the quality of a specimen.

Describe actions that ensure safety during performance of a

venipuncture.

Principle: Routine venipuncture is an excellent course for novice or

experienced phlebotomists. Important aspects of venipucture include; blood
collection procedures, the recommended order of draw, proper patient
identification, acceptable and unacceptable sites for blood collection, and
variables in collection procedures that can affect the accuracy of the blood
test.
Introduction:
Venipuncture evolved from the practice of phlebotomy. The word phlebotomy
is derived from two Greek words referring to veins and cutting; thus
phlebotomy can be defined as the incision of a vein for bloodletting or
collection.
Material:
1. Safety Needles or Butterfly needles
2. Syringes
3. Blood Collection Tubes. Tubes with different additives are used for
collecting blood specimens for specific types of tests.
(The color of the rubber stopper is used to identify these additives)
4.

Tourniquets

5. Antiseptic. Individually packaged 70% isopropyl alcohol wipes.

6. Gauze or cotton balls.

Safety/Precaution
1.

A lab coat or gown must be worn during blood collection procedures.

2.

Needles and hubs are single use and are disposed of in an appropriate
'sharps' container as one unit.

3.

Gloves are to be discarded in the appropriate container immediately

after the phlebotomy procedure.

4.

All other items used for the procedure must be disposed of according
to proper biohazardous waste disposal policy.

5.

Contaminated surfaces must be cleaned with freshly prepared 10%

bleach solution.

6.

In the case of an accidental needle stick, immediately wash the area

with an antibacterial soap, express blood from the wound and contact
your lecturer/MLT

Procedure:
VENIPUNCTURE PROCEDURE USING A SYRINGE:
1.
Place a sheathed needle or butterfly on the syringe.
2.
Remove the cap and turn the bevel up.
3.
Pull the skin tight with your thumb or index finger just below the
puncture site.
4.
Holding the needle in line with the vein, use a quick, small thrust to
penetrate the skin
and vein in one motion.
5.
Draw the desired amount of blood by pulling back slowly on the syringe
stopper.
6.
Release the tourniquet.
7. Place a gauze pad over the puncture site and quickly remove the
needle.
8. Immediately apply pressure. Ask the patient to apply pressure to the
gauze for at least
two minutes.
9. When bleeding stops, apply a fresh bandage, gauze or tape.
10. Transfer blood drawn into the appropriate tubes as soon as possible
using a
needleless.
11. Dispose of the syringe and needle as a unit into an appropriate sharps
container.

Venipuncture: Blood Draw Procedure

TROUBLESHOOTING HINTS FOR VENIPUNCTURE

If a blood sample is not attainable:

Reposition the needle.

Ensure that the collection tube is completely pushed onto the back of
the needle in the hub.

Loosen the tourniquet.

A patient should never be stuck more than twice unsuccessfully by a

phlebotomist.
The supervisor/lecturer should be called to assess the patient.

REFERENCES
NCCLS: Procedure for the Collection of Diagnostic Blood Specimens by
Venipuncture; Approved Standard, Sixth Edition, Vol 27, No 26 (H3-A6), 2007
Nursing Procedure Manual. Lippincott Online with addenda, 2004.

PRACTICAL 5(b): INTRAVENOUS FLUID INFUSION

Objective:
To expose the procedure and skills associated with intravenous fluids
and infusions
To discuss equipment used to setting up the infusions
To compare common types of IV fluid
Introduction:
Intravenous Fluid
Intravenous fluids are chemically prepared solutions that are administered to
the patient who is dehydrated, severely cramping, in shock, or as a part of
emergency medical care in a potentially life-threatening situation. They are
tailored to the bodys needs and used to replace lost fluid and/or aid in the
delivery of IV medications.
Once the prescribed intravenous fluid is selected by the sending institution's
medical staff, the I.V. Tubing should be inspected to confirm there are no air
bubbles and the container should be inspected for cracks or tears, foreign
matter, cloudiness, precipitation and any other sign of contamination. Should
any of this be apparent, the fluid must not be used. The fluid must not have
reached its expiry date. The expiry date is printed on the fluid container.
The most commonly used IV fluid are:
a) NORMAL SALINE (N/S) - An isotonic crystalloid solution that contains
sodium chloride (salt) as the solute, dissolved in sterile water (solvent).The
specific concentration for normal saline solution is 0.9%.
b) RINGERS LACTATE (Lactated Ringers) - An isotonic crystalloid
solution containing the solutes sodium chloride, potassium chloride, calcium
chloride, and sodium lactate, dissolved in sterile water (solvent).
c) D5W (5% Dextrose in water) - A carbohydrate solution that uses
glucose (sugar) as the solute dissolved in sterile water. Five percent dextrose
in water is packed as an isotonic solution but becomes hypotonic once in the
body because the glucose (solute) dissolved in sterile water is metabolized
rapidly by the bodys cells.
Solutions are available in volumes of 50, 100, 250, 500, 1000ml bags and are
sealed in an outer plastic wrap to help prevent contamination.

Equipment preparation
1. Hands must be washed and gloves should be worn.
2. Open an intravenous fluid container ensure its sterility, clarity, and
expiry date.
3. Mark the date and time intravenous fluid container was opened.
4. Hang container on a pole.
5. Remove administration set from wrapper.
6. Close clamp on administration set.
7. Attach extension set to administration set (if applicable).
8. Insert administration set drip chamber into intravenous fluid container
where indicated.
9. Squeeze and release drip chamber until half full of solution.
10.
Release clamp to fill tubing with solution.
11.
Raise end of tubing to avoid air bubbles.
12.
Invert check valve to clear and allow valve to function.
13.
Clamp tubing.
14.
The infusion should be labeled with:
the patients name and number;
the name and amount of additives;
the date and time prepared;
infusion expiry date / time;
route, diluents and final volume;
prepared by and checked by.
Procedure:
1. Ensure patient meets criteria.
2. Explain procedure to patient.
3. Select site antecutibal fossa preferred for trauma or unstable
patients.
4. Apply tourniquet.
5. Clean area over site with alcohol swabs.
6.
Prepare intravenous needle with cannula, adhesive tape, and
dressing.
7. Draw skin taut over vein.
8. Insert intravenous needle with cannula, bevel up, through the skin
into the vein.
9. Carefully remove needle, leaving cannula in vein.
10. Once vein successfully cannulated, attach intravenous tubing to
cannula.
11. Open clamp to ensure flow of intravenous fluid and apply dressing to
site.
13. Anchor intravenous tubing using tape.

14. Label site with date, type and size of cannula, and name of personnel
carrying out
procedure.
15.
Regulate rate of infusion.
16.
Intravenous Infusion Set

Fluid

Screw clamp (for flow rate adjustment)

Y injection site

Slide Clamp (For temporary flow interruption)

Piercing Pin
Protective Flange

Needle & catheter

Needle adapter

Drip chamber

PRACTICAL 5(C): BLOOD GLUCOSE TEST BY USING

GLUCOMETER
PRINCIPLE

Glucose determination was done to detect level of glucose in patient. It is

important to see whether the level of glucose can cause consequences to the
patient or not. It is either hyperglycemia or hypoglycemia. When
hyperglycemia occurs it is condition which the level of blood glucose is
higher than normal range, while hypoglycemia is low blood glucose. Both
conditions can give consequences to the patient.
Blood sugar monitoring devices called glucometer provide with instant
feedback and let know immediately what the blood sugar is. This can give
valuable information about whether the blood sugar is too low, too high or in
a good range.
MATERIAL

Alcohol prep pad

Lancet
Test strip
Glucometer

PROCEDURE
1. First, set out the glucometer, a test strip, a lancet and alcohol prep
pad.
2. Wash your hands to prevent infection.
3. Decide where you are going to obtain the blood from, usually a finger.
Some of the newer monitors let you use your forearm or another less
sensitive place.
4. Sometimes it helps to warm your hands to make the blood flow easier.
You can rub your hands together briskly or run them under warm
water.
5. Turn on glucometer and place a test strip in the machine when the
machine is ready. Watch the indicator for placing the blood to the strip.

6. Make sure your hands is dry and wipe the area youve selected with an
alcohol prep pad and wait until the alcohol evaporates.
7. Pierce your finger tip on the soft, fleshy pad and obtain a drop of blood.
He type of drop of blood is determined by the type of strip you are
using (some use a hanging drop of blood versus a small drop for
strips that draw blood in with a capillary action).
8. Place the drop of blood on or at the side of the strip.
9. The glucometer will take a few moments to calculate the blood sugar
reading. Follow your doctors orders for whatever blood sugar reading
you get.
10.
You may use the alcohol prep pad to blot the site where you drew
the blood if it is still bleeding.
11.
Write down your result. Keeping a record makes it easier for you
and your doctor to establish a good treatment plan. Some glucometers
can store your results in a memory, for easier record keeping.

EXPECTED RESULT

Diagnosis

Diagnosis blood glucose concentrations (mmol/L)

Sample
Plasma glucose
concentration

Normal

Fasting

<6.1

Impaired fasting
glycaemia

Fasting

>6.1 and <7.0

2h post glucose

<7.8

Fasting

<7

2h post glucose

>7.8 and >11.1

Fasting

>7.0

2h post glucose

>11.1

Impaired glucose
tolerance

Diabetes mellitus

RESULT

DISCUSSION
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CONCLUSION
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