COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Safety
Each professional must be “safety conscious”
at all times!
 Electric shock (use of electrical
machines), toxic vapors (coming from
reagents we are using), compressed
gases (LPG or liquid nitrogen),
flammable liquids, radioactive
material, and corrosive substances;
 Mechanical trauma (cuts from sharp
equipments that we use);
 Poisons; and
 Inherent risks of handling biologic
materials (samples that we receive or
process).
 Unsafe acts (not always recognized by
personnel).
 Unsafe environmental conditions.
o Slippery floor, improperly
ventilated area, or small
working space.
 First rule of self-protection
 alertness at all times.
 Stay informed.
 Use common sense.
 LISTEN to any instructions.
 Pertains to the proper way of thinking
or behavior inside the laboratory.
1. Laboratory safety necessitates the
effective control of all hazards that exists in
the clinical laboratory at any time.
2. Safety begins with the recognition of
hazards and is achieved through:
 Application of common sense.
 Safety-focused attitude.
 Good personal behavior.
 Good housekeeping in all laboratory
work and storage areas.
 Continual practice of good laboratory
technique.
3. Inexperience may cause some accidents;
others are results of ignoring known risks,
haste carelessness, fatigue or mental
preoccupation.
4. Preventive Measures:
 Annual safety reviews.
o Every year the safety
equipments in the laboratory
are inspected.
 Safety drills.
o Earthquake drill and fire drill.
 General consciousness.
o Use of common sense.
 Appropriate orientation to safety rules.
 Safe work environment.
o Obligation of the employers.
o According to OSHA, it is the
responsibility of the employers
to give safety work
environment to its employees.
 Center for Disease Control (CDC)
 Universal Precautions (1987)
o Blood and body fluid
precautions should be
consistently used for all
patients.
 Potentially infectious materials:
o Body fluids semen, vaginal
secretions, amniotic fluid,
saliva, tears, CSF, urine and
breast milk.
o Unfixed tissues, organs or blood
slides.
 Precautions:
o Appropriate barriers (gloves,
gowns or laboratory coats).
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Safety
o Appropriate engineering
controls.
 Universal Practices:
o Wearing of gloves  reusing is
not allowed.
o Handwashing.
o Laboratory coats  on site.
 Laboratory coats must
be worn only on the
laboratory.
o Prohibited: eating, drinking,
smoking, applying cosmetics,
touching contact lenses.
 Inactivation:
o Heat sterilization (250˚C for 15
minutes).
o Ethylene Oxide (450-500 mg/L
at 55-60˚C).
o 2% Glutaraldehyde.
o 10% hydrogen peroxide.
o 5.25 hypochlorite (bleach).
o 10% (v/v with tap water) of
common household bleach)
 HBV (10 minutes), HIV (2
minutes).
 Vaccination against HBV (medical
technologists, phlebotomists,
pathologists).
 Appropriate signs to identify hazards.
 Developed specifically for use in the
laboratory.
 Safety showers, eyewash stations, fire
extinguishers  periodically tested
and inspected.
 Mechanical pipetting device must be
used to manipulate liquids.
1. All samples and other body fluids
should be transported, handled, and
processed using strict precautions.
2. Gloves, gowns and face protection
must be used if splash or splattering is
likely to occur.
3. Specimens should be “capped” during
centrifugation.
4. Any blood, body fluid, or other
potentially infectious material spill
must be cleaned up.
 Wear appropriate protective
equipment.
 Use mechanical devices to pick-up
broken glass or other sharp objects.
 Absorb spills with paper towels,
gauze pad, or tissue.
 Clean the spill site using a common
aqueous detergent.
 Disinfect the spill site using approved
disinfectant or 10% bleach using
appropriate contact time.
 Dispose all materials in appropriate
biohazard containers.
5. OSHA Blood-Borne Pathogens
standard requires written “Exposure
Control Plan”.
6. Categories of Exposure:
a. Category I – daily exposure to
blood and body fluids.
b. Category II – regular exposure to
blood and body fluids.
c. Category III – no exposure to blood
and body fluids.
o Employers must offer HBV to all
personnel (Category I and II).
7. Biological Safety Cabinets should be
installed to facilitate manipulations of
infectious materials.
8. Safety against exposure to toxic
chemicals.
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Safety
a. Hazard Communication  Special information section
Standard:
1) Hazard communication program
2) Chemical Hygiene Plan
3) Inventory of hazardous
substances
b. Toxic chemicals
(Communication):
1) Labelling of containers
2) Information and training d. NFPA hazard labelling system
3) Program of Hazard

e. Fume hoods
communication o Reagent preparation.
c. MSDS  properties and effects of
each chemical.

 Product name and identification

 Hazardous ingredients
 Permissible exposure limit (PEL)
 Physical and chemical data
 Health hazard data and
carcinogenic potential
 Primary routes of entry
 Fire and explosion hazards o Required to expel noxious and
 Reactivity data hazardous fumes from
 Spill and disposal procedures chemical reagents.
 PPE recommendations
 Handling
 Emergency and first aid procedures
Flammable/Combustible Chemicals
 Storage and transportation
precautions 
 Chemical manufacturer’s name, temperature at which sufficient vapor
address, and telephone number is given off to form an ignitable mixture
with air.

Corrosive Chemicals

 Injurious to the skin or eyes by direct

contact or to the tissue of the

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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Safety
respiratory and gastrointestinal tracts
if inhaled or ingested.
Reactive Chemicals
 Spontaneously explode or ignite or that
evolve heat or flammable or explosive
gases.
Carcinogenic Chemicals
1. Lock-out or tag malfunctioning
electrical or mechanical equipment
until serviced.
2. Know how to knock a shocked person
loose using a non-conductive material.
 Use only explosion-proof equipment in
hazardous atmospheres.
 Be particularly careful when operating
highvoltage equipment, such as
electrophoresis apparatus.
 Use only properly grounded equipment
(threeprong plug).
 Check for frayed electrical cords.
 Promptly report any malfunctions or
equipment.
 Do not work on “live” electrical
equipment.
 Never operate electrical equipment
with wet hands.
 Know the exact location of the
electrical control panel for the
electricity to your work area.
 Use only approved extension cords and
do not overload circuits. (Some local
regulations prohibit the use of any
extension cord).
 Have ground checks and other periodic
preventive maintenance performed on
equipment.
 Radiation safety policy should include
environmental and personnel
protection.
 All areas where radioactive materials
are used or stored must be posted with
caution signs, and traffic in these areas
should be restricted to essential
personnel only.
 Records must be maintained for the
length of employment plus 30 years.
 Radiation monitoring utilizes film
badge or survey meter  maximum
permissible dose is 5000 mrem/year
whole body.
 The wipe test (leak test) involves wiping
laboratory surfaces with moistened
absorbent material and the radiation
contained in each wipe is counted.
 Danger of fire
 Explosion
 Asphyxiation
 Mechanical injuries
 Know the gas that you will use.
 Store tanks in a vertical position.
 Keep cylinders secured at all times.
 Never store flammable liquids and
compressed gases in the same area.
 Use the proper regulator for the type of
gas in use.
 Do not attempt to control or shut off
gas flow with the pressure relief
regulator.
 Keep removable protection caps in
place until the cylinder is in use.
 Make certain that acetylene tanks are
properly piped (the gas is incompatible
with copper tubing).
 Do not force a “frozen” or stuck
cylinder valve.
 Use a hand truck to transport large
tanks.
 Always check tanks on receipt and
then periodically for any problems such
as leaks.
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Safety
 Make certain that the cylinder is
properly labeled to identify the
contents.
 Empty tanks should be marked
“empty”.
 Liquid nitrogen  most widely used
cryogenic fluids (liquefied gases) in the
laboratory.
 It may cause fire or explosion,
asphyxiation, pressure buildup,
embrittlement of materials, and tissue
damage similar to that of thermal
burns.
 Centrifuges  must be balanced to
distribute the load equally.
o Never open the lid until the
rotor has come to a complete
stop.
o Safety locks on equipment
should never be rendered
inoperable.
 Glass beads – help eliminate
bumping/boil over when liquids are
heated.
 Infectious sharps - disposed in OSHA-
approved containers.
4 Basic Waste Disposal Technique
 Flushing down the drain
 Incineration
 Landfill burial
 Recycling
Chemical Waste
 Flush water-soluble substances down
the drain with large quantities of water.
 Strong acids and bases should be
neutralized before disposal.
 Foul smelling chemicals should never
be disposed down the drain.
 Flammable solvents  collected in
approved containers.
 Flammable material  specially
designed incinerators.
 Solid chemicals  landfill.
Radioactive Waste
 Depends on the type of waste (soluble
or non-soluble), its level of
radioactivity, and the radiotoxicity and
half-life of the isotopes involved.
Biohazardous Waste
 Medical waste  special waste from
health care facilities.
o Solid waste that, if improperly
treated or handled, “may
transmit infectious diseases”.
o Blood and blood products,
microbiologic waste, pathologic
waste and sharps.
 Steam sterilization, incineration,
thermal inactivation, burial, chemical
disinfection, encapsulation in a solid
matrix.
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Equipment
 Used for mixing, transferring or
measuring.
 Acts as a container in the laboratory.
 Some of the equipments are made up
of glass and we call them as a
glassware, can also be made up of
plastic, metal, or ceramics.
 Pipettes are glass or plastic tubes,
usually open at both ends, which are
used to transfer specific amounts of
liquid from one container to another.
 They are usually used for volumes
between 1 and 100 milliliters.
Depends on the amount of liquid needed to
wet the interior surface of the ware and the
amount of any residual liquid left in the pipet
tip:
I. Base on Design
a. To contain
o Types of pipette that can
aspirate or contain a specific
volume of liquid.
o Contains a particular volume
but does not dispense the exact
volume.
o Rinse out pipet (Diluting fluid)
o A small amount of fluid will
cling to the inside wall of the
pipet.
Ex: Sahli-hemoglobin, Lang-
levy, WBC pipette, and RBC
pipette
Sahli-
hemoglobin
Lang-levy
b. To deliver
o They aspirate or contain but at
the same time deliver the exact
volume of liquid.
o Dispense the amount of volume
indicated.
o Designed to be drain by gravity.
o Must be held vertically and the
tip placed against the side of
the container and must not
touch the liquid in it.
o A small amount of fluid will
remain in the tip of the pipette.
o Meet requirements of transfer
pipets.
Ex: Mohr, Serologic, Volumetric
transfer pipets
II. Base on Drainage
a. Blow Out
o It has a continuous etched ring
or two small continuous rings
located near the top of the
pipet.
o Last drop should be expelled
into receiving vessel.
 An aspirator bulb is
needed to remove the
remaining liquid.
o The frosted band should not be
confused with thicker colored
rings or colored dots, which are
a manufacturer’s code for the
maximum volume of the
pipette.
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Equipment
 Remember, only Ex: Serologic, Mohr,
blowout a serological Bacteriologic, Ball, Kolmer, or
pipette if it has a frosted Kahn, and Micropipet (pipette
band or two thin rings. that can aspirate and
Ex: Serologic, Ostwald Folin dispense liquid between 0.1 to
1 ml or less than 1 ml)

b. Transfer
o Ostwald-Folin Pipet
 For viscous fluids.
b. Self-Draining  Blow out.
o Allows the content to drain by  Has bulb near the tip.
gravity.
o The tip of the pipet should not
be in contact with the
accumulating fluid in the
receiving vessels during
drainage except mohr pipet.
Ex: Volumetric, Mohr
III. Base on Use o Volumetric Pipet
a. Measuring or Graduated  Has the greatest degree
o Deliver the amount of liquid of accuracy and
contained between two precision.o
calibration marks.  Used to deliver a single
o Not calibrated with sufficient specific volume of liquid,
tolerance to use in measuring usually between 1 and
standard or control solutions. 100 ml.
o Measuring pipettes are divided  Shaped like rolling pins
into: with a large belly, one
 Mohr Pipettes (the blunt end, the neck, and
graduations on these one tapering end, the
always end before the tip.
tip) o Pasteur Pipet
 Do not have calibration
marks.

 Serological Pipettes
(the graduation marks
continue to the tip)

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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Equipment
 Use to transfer solutions
without consideration of
Dispenser and Dilutor (Automatic Pipet)
a specific volume.
 The dilutor often combines sampling
and dispensing functions.

Pipet Bulbs

 A pipette bulb is used to draw liquid up

into the pipette. There are many types
of pipette bulbs:
o Types of Automatic Pipet 1. Rubber Bulb
a. Air Displacement 2. Pipet Filler
 Relies on piston for suction creation to 3. Pipet Aid
draw the sample into a disposable tip 4. Pipet Pumper
that must be changed for use.
 The piston does not come in contact
with the liquid. Rubber
b. Positive-Displacement Bulbs
 Operates by moving the piston in the
pipet tip.
 Like syringe.
 Doesn’t require different tips.
 Rinsing and blotting between samples
maybe required. Pipet
c. Dispenser and Dilutor/Dispenser Pumper
 Obtain the liquid from common
reservoir and dispense it repeatedly.
1. Bottle-top
2. Motorized
3. Handheld
4. Attached to a dilutor
Pipet Aid

Pipet
Pumper

Legend: (A&C) – air displacement automatic

pipet; (B&D) – positive displacement automatic
pipet
1. Hold the pipette about 8 cm below the
mouthpiece with one hand. Then with your
other hand squeeze the bulb and touch the
opening to the mouth of the pipette.
2. Insert no more than one-half cm of the
pipette into the bulb.

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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Equipment
3. Place the tip into the colored liquid and  Clear solutions.
slowly release the pressure on the bulb.
Upper Meniscus
4. The liquid will be drawn up into the pipette
and will form a curved surface against the  Colored or viscous solutions.
glass.
5. This surface is called the meniscus. Pull the
bottom of the meniscus up about 1 cm past
the desired level.
6. Then quickly, but carefully, remove the
bulb as you slip your free index finger over
the tip of the mouthpiece hole.
 NEVER USE YOUR THUMB--your index
finger will allow better control and will
also enable you to hold other items
with your free fingers when necessary.  Calibration
7. Then with your finger still on the end of the o Glassware or other apparatus
pipette, gently lift the pipette out of the used in quantitative
solution. measurement is checked to
8. Then raise your finger just enough to allow determine its exact volume.
the bottom of the meniscus to line up with
the desired graduation mark. YOU  Calibrated by weight using distilled
SHOULD OBSERVE THE MENISCUS AT water and analytic balance.
EYE-LEVEL WHILE DOING THIS.  Volumetric flasks
 When the meniscus is at the desired o Calibrated to hold one exact
level, touch the tip of the pipette to the volume of liquid (TC).
inside of the container holding the
colored water, to remove any drops of
liquid on the end of the pipette. Now,
there is precisely (0.645 + 0.001) ml of
colored water in your pipette.
9. Keeping your finger on the end of the
pipette, wipe the sides of the pipet with
tissue paper & gently move it to the waste
container.  Erlenmeyer Flask
10. Touch the tip to the inside of the tilted o Designed to hold different
container, lift your finger off the end and volumes rather than one exact
allow the liquid to drain out of the pipette. amount.
11. Hold the pipette in this position for a few
seconds after it stops draining wipe the
pipet again before disposal or cleaning the
pipet.

 Curvature in the top surface of the

liquid. Pipette should be held that the  Beaker
calibration mark is at the eye level. o Hold different volumes rather
than one exact amount.
Lower Meniscus
o Griffin beakers.

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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Laboratory Equipment
o Berzeleus beakers. Surface  For flat surface
Thermometers such as oven or
incubator.
2. Electronic Thermometer/ Thermistor
Probe
o Advantage:
 Size and millisecond
 Response time
 Graduated Cylinder
o Disadvantage:
o Used to measure liquid when
 Expensive
high degree of accuracy is not
3. Digital Thermometer
essential.
o Calibrated to deliver.
o Can be used to measure
specified volume of liquid.

 Thermometer
1. Liquid-in-glass
o Use color liquid or mercury
encased in plastic or glass
material with bulb at one end
and a graduated stem.
o Usually measures temperature
between 20 ̊C to400 ̊C.

Partial Immersion  Used for

measuring temp
in units such as
heating blocks
and water bath.
Should be
immersed to the
proper height as
indicated by the
continuous line
etched in the
thermometer.
Total Immersion  Used for
refrigeration
application.

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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Collection of Specimen
Antecubital Space
 Located where the elbow ends.
Arterial Blood Gas (ABG)
 Procedure that involves taking blood
from an artery.
Artery
 Blood vessel that carries richly
oxygenated blood from the heart to the
body tissues.
Capillaries
 Tiny blood vessels that connect the
smallest arteries to the smallest veins.
Gauge
Hematoma
 Loss of blood from a vessel.
Hemoconcentration
 Increase in the concentration of formed
elements in the blood caused by lack of
fluid in the blood.
Hemolysis
 Destruction or lysis of red blood cells.
Lumen
 Cavity or channel within the tubular
organ, such as blood vessel.
Phlebotomy
 Act of entering a vein with a needle for
the purpose of obtaining venous blood.
Thrombosis
 Blood clot in a vessel.
Tourniquet
 Device often a rubber band at least ½
inch wide, that reduces flow of blood in
the veins and allows the veins become
prominent.
Vacuum Tube system/Vacutainer System
 System used for blood collection; uses
a special disposable needle, a holder,
and blood collection tubes that have
little or no air.
Vein
 Blood vessels that carries
deoxygenated blood from the tissues
back to the heart.
Venipuncture
 Procedure where a needle is inserted
into a vein to obtain a venous blood
sample.
 If a fasting specimen is required,
confirm that the fasting order has been
followed.
 Always read the request first to see
whether the examination is complete
or incomplete, to determine the
amount of blood needed.
 Do not extract blood from patients
while they are receiving intravenous
medication because these solutions
may influence the chemical analysis.
 Never draw out the syringe without
removing first the tourniquet to avoid
hematoma.
 Avoid prolonged application of
tourniquet to avoid
hemoconcentration.
 The blood specimen collected with
anticoagulants must be well mixed to
prevent coagulation.
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Collection of Specimen
a. Whole blood
o Plasma and formed elements
(unclotted).
b. Serum
o Liquid portion of clotted blood.
o Contains albumin and globulin.
o Preferred specimen in clinical
chemistry because of the
following reasons:
 Certain substances like
the lipemia clearing
factor (LPL) are co
precipitated during
clotting.
 Clearer than plasma.
 There is no potential
interference produced
by anticoagulant.
c. Plasma
o Liquid portion of unclotted
blood.
o Contains albumin, globulin and
fibrinogen.
o Advantages of plasma over
serum:
 There is no delay
necessary in obtaining
plasma by
centrifugation of the
whole blood.
 Usually a greater
volume of plasma than
serum is obtainable
from a given volume of
blood.
 24-hr sample.
 Random sample.
 Timed sample.
 CSF (cerebrospinal fluid).
 Other specimens like ascitic fluid,
peritoneal fluid, etc.
 Fasting
o “Nothing by mouth”.
o Generally 6 -14 hours.
 Random
 24-Hour
 Postprandial
 Macromethod
 Micromethod
 Ultramicromethod
 Nanoliter method
a. Transient or immediate effects (within
the hour)
o Elevated lactate
o Elevated alanine
o Elevated fatty acids
b. Long lasting effects
o Increased activity of:
 AST (SGOT)
 CPK (CK)
 LDH
 ALD (Izoenzyme A)
c. Long term effects
o Elevated concentration of sex
hormones
 Testosterone
 Luteinizing hormone
 Sex hormone binding
globulin
o Elevated concentration of
steroids
 Generally 6-14 hours.
 Elevated blood glucose, potassium, and
lipids like cholesterol and neutral fats
are seen in patients not under NPO
and the reverse is true with inorganic
phosphorous.
 Prolonged fasting has been associated
with elevations in serum bilirubin,
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Collection of Specimen
plasma triglycerides, glycerol, free fatty
acids and a decrease in plasma
glucose.
 Any diet immediately increases
potassium and triglycerides
 2-4 hours after fatty meal – increases
ALP activity.
 High protein diet – increases urea,
ammonia and urates with no
significant increase in creatinine.
 Increase turbidity or lactescence –
triglycerides level exceeds 4.6 mmol/L
(4.0 g/L).
 Changes in position result to efflux of
filterable substances from the
intravascular space to the interstitial
fluid spaces.
 Non-filterable substances increase in
concentration.
 One-minute application
 Prolonged application results to:
o Venous stasis
o Anaerobiasis
a. Acute effects
o Increase in plasma NEFA
concentration.
o Increase in plasma
cathecolamines and serum
cortisol (due to nicotine) –
affects the leukocyte peripheral
count.
 Increase in neutrophils
 Increase in monocytes
 Increase in eosinophils
b. Chronic effects
o Increase in total WBC count.
o Increase in blood hemoglobin
values (carboxyhemoglobin).
o Increase in mean corpuscular
Volume (MCV).
 Increases plasma concentration of
lactate, urate, acetate and
acetaldehyde.
 Increases gamma glutamyl transferase
concentration.
 Affects hormone secretion.
 Results in hyperventilation leading to a
disturbance in acid-base balance in
the blood.
 Increases serum lactate.
 Increases plasma free fatty acids.
 Drugs
o Antacids
 For micromethod, ultramicromethod
and nanoliter method.
 The method of choice for:
o Pediatric infant
o Geriatrics
o Adults who are:
 Sites of Puncture
o Palmar surfaces of fingertips
o Plantar heel surface in infants
o Plantar surface of the big toe
o Earlobes
 Sites to be avoided
o Edematous area (swollen with
fluid)
o Cyanotic areas (purplish blue in
color)
o Scarred area
o Traumatized area (black and
blue)
o Heavily calloused area
 Materials and Equipments
o 70% isopropyl alcohol
o Absorbent cotton sponges/balls
or swabs
o Sterile puncture instrument
(lancet)
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COLLEGE OF MEDICAL LABORATORY SCIENCE
Clinical Chemistry 1
Collection of Specimen
o Capillary tube o Jugular vein
o Saphenous vein
 Generally used for the determination of  Small saphenous vein
blood oxygen, carbon dioxide tension  Great saphenous vein
and blood pH (Blood Gas Analysis). o Veins on the dorsal portion of
 Special training is required for this the hand
procedure.  Materials needed
 Tourniquet is not required. o 70% alcohol
 After removing the needle, apply o Absorbent cotton or sterile
moderate pressure with 2 x 2 sterile swabs
gauze until bleeding ceases. o Tourniquet
 Insert needle (still attached to syringe) o Syringe or Vacutainer set
in stopper to prevent air from entering
needle.
 Sites of puncture  Used for blood collection only
o Radial artery  For single draw
 Most preferred site  Parts
o Femoral artery (fem tap) o Bevel
o Brachial artery o Adapter
o Scalp artery o Shaft
o Umbilical artery o Barrel
 Materials needed o Hub
o Syringe with gauge 21 needle o Plunger
with rubber cap
o Cotton (wet and dry)
o Alcohol or Iodine  Large amount of blood can be
o Heparin obtained.
 Preferred anticoagulant  Ideal for blood chemistry
o Warm towel (45˚C) determinations.
 Arterialization  It reduces the possibility of error in
o Iced water bath resulting from dilution with tissue
 Preservative for juices.
transport

 Considered to be the most commonly  Requires more time and skill.

used method of blood collection in  Requires more equipment.
Clinical Chemistry.  More complications may arise.
 Sample obtained is called venous  Hard to do on infants, children and
blood. obese individuals.
 Sites of collection
o Veins in the antecubital fossa
 Median cephalic vein –
 Assembling equipments.
 Cephalic vein –
o Prepare the materials that will
 Basilic vein –
be needed in the procedure.
o Vein of the longitudinal sinus or
 Positioning and identifying the patient.
sagittal sinus
 Preparation of the needle and syringe.
o Femoral vein
 Applying the tourniquet.
o Wrist vein
 Selecting the vein.

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 Applying the antiseptic.
 Inserting the needle.
 Releasing the tourniquet.
 Withdrawing the blood.
 Withdrawing the needle.
 Transferring the blood.
 Checking the patient wound.
 Localized hemoconcentration or
Venous stasis
 Syncope or Fainting
o Due to sudden decrease of
blood supply to the brain.
o Remedy:
 Let the patient lie down.
 Failure of blood to enter the syringe
due to:
o Excessive pull of the plunger
which collapses the vein.
o Going through the vein
reaching the musculature.
o Very small angle of entry
wherein the needle reaches
only the walls of the veins
(transfixation of the vein).
 Thrombosis of veins
o Formation of blood clots inside
the lumen of the vein due to
trauma.
 Thrombophlebitis
o Inflammation of the vein due to
thrombus as manifested by an
inflammatory reaction on the
outer skin surface.
 Hematomas
o Blue or black skin discoloration
commonly due to repeated
trauma or puncture of the
veins.
 Serum Hepatitis
 AIDS
 This must be avoided because of the
following reasons:
o Most constituents, such as
SGOT, LDH, Acid Phosphatase
and Potassium are present in
large amount in erythrocytes
o Invalidates determination due
to color changes
o May directly interfere in a
chemical determination by
inhibiting an enzyme such as
lipase.
o Hemoglobin may interfere with
the diazotization of bilirubin.
 Hemolysis can be prevented By
o Proper collection of blood
 Venous stasis
 Moisture
 Excessive traction
 Small lumen needles
 Air leak
o Proper transfer of blood
 Expelling of blood
through the needle
 Shaking
o Separation of blood
 Freezing
 Over centrifuge
 This is caused by transient rise in
chylomicrons following a meal
containing fat.
 It causes interference with large
number of chemical analyses because
of turbidity.
 It disturbs the following investigations
particularly strongly:
o Amylase
o Bilirubin
o Protein
o SGOT
o SGPT
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 This may occur through dilution or

evaporation.
 The sources of the errors are:
o Use of syringe and needles
rinsed in saline solution.
o Use of liquid form of
anticoagulant.
o Allowing the blood to stand in
open container.
o Centrifugation of blood
specimens in open containers.

 Bacterial changes
o Such as formation of ammonia
from urea.
 Enzymatic changes
 Blood gas changes
 Extravascular interchange
o This is due to movement of
substance between the cells
and plasma or serum, like K,
water and chloride.
 Changes in lipid concentration
o Due to lipolysis.
 Changes in phosphates
o Due to hydrolysis of organic
phosphate esters.

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