lOMoARcPSD|16683526

CLINICAL CHEMISTRY 1- LABORATORY • Not calibrated with sufficient tolerance to use in measuring standard or control
solutions.
I. LABORATORY EQUIPMENT • The two major type is the serologic and mohr pipette
1. Serologic
BASIC LABORATORY EQUIPMENT AND SUPPLIES
2. Mohr
A. PIPETS: 3. Bacteriologic
• PIPETTES are glass or plastic tubes, usually open at both ends, which are used 4. Ball, kolmer, or kahn
to transfer specific amounts of liquid from one container to another. 5. Micropipet
• They are usually used for volumes between 1 and 100 milliliters.
2. TRANSFER- more accurate than measuring pipette
CLASSIFICATION OF PIPETS 1. Volumetric- calibrated for one specific volume
2. Ostwald- folin - calibrated for one specific volume
• Depends on the amount of liquid needed to wet the interior surface of the ware and 3. Pasteur pipets
the amount of any residual liquid left in the pipet tip: 4. Automatic macropipets or micropipets

1. BASE ON DESIGN: MEASURING PIPETS

• To contain (tc)
• To deliver (td) MEASURING PIPETTES ARE DIVIDED INTO:
2. BASE ON DRAINAGE:
• MOHR PIPETTES
• Blow out
• The graduations on these always end before the tip
• Self-draining
• SEROLOGICAL PIPETTES
3. BASE ON USE:
• The graduation marks continue to the tip
• Measuring / graduated
• Volumetric / transfer OSTWALD - FOLIN PIPET (TRANSFER PIPET)

TC – TO CONTAIN PIPET • FOR VISCOUS FLUIDS such as protein and whole blood standards
• Mouthpiece has a frosted ring
• Contains a particular volume but does not dispense the exact volume
• BLOW OUT
• Rinse out pipet (diluting fluid) and not always used in clinical chemistry and used
• HAS BULB NEAR THE TIP
in diluting highly viscous fluid such as whole blood
• A small amount of fluid will cling to the inside wall of the pipet.
➢ Sahli pipette are smaller but similar to Oswald folin and is used to make dilution of
• Example:
whole blood for hematology
o Sahli-hemoglobin-used for measurement of hemoglobin and clot retraction
➢ HAS THE GREATEST DEGREE OF PRECISION
o Lang-levy
VOLUMETRIC PIPET (TRANSFER PIPET)
TD – TO DELIVER PIPET
• Used to deliver a single specific volume of liquid, usually between 1 and 100
• Dispense the amount of volume indicated
ml.
• Designed to be drain by gravity
• Shaped like rolling pins with a large belly, one blunt end, the neck, and one
• Must be held vertically and the tip placed against the side of the container and
tapering end, the tip.
must not touch the liquid in it.
• Volume calibration mark is etched at the neck of the pipette and no etched
• A small amount of fluid will remain in the tip of the pipette
ring at the mouth piece
• Meet requirements of transfer pipets
• To deliver and self-draining
• Ex:
• Cylindrical glass bulb at the center
o Mohr
o Serologic PASTEUR PIPET (TRANSFER PIPET)
o Volumetric transfer pipets
• Do not have calibration marks
CLASSIFICATION – DRAINAGE CHARACTERISTICS • Use to transfer solutions without consideration of a specific volume

1. BLOW OUT TYPES OF AUTOMATIC PIPET

• Has a continuous etched ring or two small continuous rings located near
the top of the pipet 1. AIR DISPLACEMENT
• Last drop should be expelled into receiving vessel • Relies on piston for suction creation to draw the sample into a disposable tip
• Ex: serologic, ostwald folin that must be changed for use.
2. SELF-DRAINING • The piston does not come in contact with the liquid
• Without etched ring to the top 2. POSITIVE-DISPLACEMENT
• Highly accurate pipettes • Operates by moving the piston in the pipet tip. Like syringe.
• Allows the content to drain by gravity • Doesn’t require different tips
• The tip of the pipet should not be in contact with the accumulating fluid • Rinsing and blotting between samples maybe required
in the receiving vessels during drainage except mohr pipet 3. DISPENSER AND DILUTOR/DISPENSER
• Ex: volumetric, mohr • Obtain the liquid from common reservoir and dispense it repeatedly.
1. BOTTLE-TOP
BLOW OUT PIPET 2. MOTORIZED
3. HANDHELD
• The frosted band should not be confused with thicker colored rings or colored
4. ATTACHED TO A DILUTOR
dots, which are a manufacturer’s code for the maximum volume of the pipette.
• Remember, only blow- out a serological pipette if it has a frosted band or two DISPENSER AND DILUTOR (AUTOMATIC PIPET)
thin rings.
• The dilutor often combines sampling and dispensing functions
CLASSIFICATION -BASED ON USE
PIPET BULBS/ASPIRATOR BULB
1. MEASURING OR GRADUATED
• Deliver the amount of liquid contained between two calibration marks and can • A pipette bulb is used to draw liquid up into the pipette. There are many types
deliver multiple measurements of pipette bulbs:
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1. RUBBER BULB • Berzeleus beakers

2. PIPET FILLER
3. PIPET AID Graduated cylinder
4. PIPET PUMPER
• Tall flask with multiple volume graduation
PROCEDURE ON USING THE PIPET • Used to alliquote volume of a fluid and to determine the volume of a fluid
• Used to measure liquid when high degree of accuracy is not essential
1. Hold the pipette about 8 cm below the mouthpiece with one hand. Then with • Calibrated to deliver
your other hand squeeze the bulb and touch the opening to the mouth of the • Can be used to measure specified volume of liquid
pipette. • Has higher degree of accuracy than the Erlenmeyer flask since the volume
2. Insert no more than one-half cm of the pipette into the bulb. markings are graduated to at least 5% of the total volume
3. Place the tip into the colored liquid and slowly release the pressure on the
bulb Thermometer
4. The liquid will be drawn up into the pipette and will form a curved surface
1. Liquid-in-glass
against the glass
• Use color liquid or mercury encased in plastic or glass material with
5. This surface is called the meniscus. Pull the bottom of the meniscus up about 1
bulb at one end and a graduated stem
cm past the desired level
• Usually measures temperature between 20 ̊c to 400 c̊
6. Then quickly, but carefully, remove the bulb as you slip your free index finger
over the tip of the mouthpiece hole Liquid-in-glass (3 types)
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 A. Partial immersion- used for measuring temp in units such as heating blocks
7. Then with your finger still on the end of the pipette, gently lift the pipette out and water bath. Should be immersed to the proper height as indicated by the
of the solution continuous line etched in the thermometer
8. Then raise your finger just enough to allow the bottom of the meniscus to line B. Total immersion- used for refrigeration application.
up with the desired graduation mark. You should observe the meniscus at eye- C. Surface thermometers- for flat surface such as oven or incubator
level while doing this.
- When the meniscus is at the desired level, touch the tip of the pipette to 2. Electronic thermometer/ thermistor probe
the inside of the container holding the colored water, to remove any drops • Advantage: size and millisecond
of liquid on the end of the pipette. Now, there is precisely (0.645 + 0.001) • Response time
ml of colored water in your pipette. • Disadvantage: expensive
9. Keeping your finger on the end of the pipette, wipe the sides of the pipet with 3. Digital thermometer
tissue paper & gently move it to the waste container.
10. Touch the tip to the inside of the tilted container, lift your finger off the end II. LABORATORY SAFETY RULES AND REGULATIONS
and allow the liquid to drain out of the pipette.
11. Hold the pipette in this position for a few seconds after it stops draining. Wipe Occupational Safety and Health Act
the pipet again before disposal or cleaning the pipet
• Public law 91-596
Meniscus - curvature in the top surface of the liquid. Pipette should be held that the • Their goal is to provide all employees with a safe work environment.
calibration mark is at the eye level. • Authorized to conduct on-site-inspection to determine whether an employer is
complying with the mandatory standards.
• Lower meniscus – clear solutions
• Upper meniscus- colored or viscous solutions SAFETY AWARENESS FOR CLINICAL LABORATORY PERSONNEL

LABORATORY VESSELS: Employer's Responsibilities

Calibration – glassware or other apparatus used in quantitative measurement is checked • Establish laboratory work methods and safety policies.
to determine its exact volume • Provide supervision and guidance to employees.
• Provide safety information, training, PPE, and medical surveillance to
National bureau of standards- Calibrated by weight using distilled water and analytic employees.
balance • Provide and maintain equipment and laboratory facilities that are free of
recognized hazards and adequate for the tasks required.
Volumetric flaks
Employee's Responsibilities
• Calibrated to hold one exact volume of liquid (tc) and has high accurate
concentration • Know and comply with the established laboratory safe work practices.
• Used in preparing solution of known volume • Have a positive attitude toward supervisors, coworkers, facilities, and safety
• Tall, slender neck and a pear-shaped body with a flat bottom and designed for training.
a single volume • Be alert and give prompt notification of unsafe conditions or practices to the
• Common volume are 250ml, 5000ml, 1000, and 2000ml immediate supervisor and ensure that unsafe conditions and practices are
corrected.
Erlenmeyer flask • Engage in the conduct of safe work practices and use of PPE.
• Designed to hold different volumes rather than one exact amount. Signage and Labeling
• Less accurate than the volumetric flask
• Conical container with multiple volume marker The National Fire Protection Association (NFPA) developed a standard hazard
• Used to dissolve a solid solute in a solution identification system (diamond-shaped, color-coded symbol), which has been adopted by
• Has a large surface area with a straight conical side many clinical laboratories.
• Used to prepare solution of known volume
• BLUE- health hazard
Beaker • RED- flammability hazard
• YELLOW- instability hazard
• Hold different volumes rather than one exact amount and has a small pour • WHITE- special hazard
pout
• Has the least accuracy
• Used to hold stock solution
• Griffin beakers
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SAFETY EQUIPMENT

All laboratories are required to have

• safety showers,
• eyewash stations, and
• fire extinguishers and to
• periodically test and inspect the equipment for proper operation.

Mechanical pipetting devices must be used for manipulating all types of liquids in the
laboratory, including water. Mouth pipetting is strictly prohibited.

Chemical Fume Hoods and Biosafety Cabinets

Fume Hoods

• required to contain and expel noxious and hazardous fumes from chemical Chemical Storage Equipment
reagents.
• Steel safety cabinets with self-closing doors are required for the storage of
• should be visually inspected for blockages.
flammable liquids
Biosafety Cabinets • Explosion-proof refrigerators may be used to store flammable materials
• Only the amount of chemical needed for that day should be available at the
• remove particles that may be harmful to the employee who is working with bench.
potentially infectious biologic specimens. • Gas cylinder supports or clamps must be used at all times,and
• Larger cylinders should be transported with valve caps on, using handcarts.
BSC CLASS
PPE and Hygiene

• Safety glasses,
• goggles,
• visors, or
• work shields - protect the eyes and face from splashes and impact
• Gloves
• rubberized sleeves
• protect the hands and arms when using caustic chemicals.
• Proper footwear is required; shoes constructed of porous materials, open toed
shoes, and sandals are considered ineffective against spilled hazardous liquids.
• Respirators may be required for various procedures in the clinical laboratory.
• Hand washing is a crucial component of both infection control and chemical
hygiene.

CLASSIFICATION OF INFECTIVE MICROORGANISM BY RISK GROUP BIOLOGIC SAFETY

• All blood samples and other body fluids should be collected, transported,
handled, and processed using universal precautions
• consistent and thorough hand washing is an essential component of infection
control.
• Antiseptic gels and foams may be used at waterless stations between washes,
but they should not take the place of an actual hand wash.
• Ideally, specimens should remain capped during centrifugation, or several
minutes should be allowed to elapse after centrifugation is complete before
opening the lid.

SPILLS

• Alert others in area of the spill.

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• Wear appropriate protective equipment. Carcinogenic Chemicals

• Use mechanical devices to pick up broken glass or other sharp objects.
• Absorb the spill with paper towels, gauze pads, or tissue. • Benzidine is a common example of a known carcinogen.
• Clean the spill site using a common aqueous detergent.
Chemical Spills
• Disinfect the spill site using approved disinfectant or 10% bleach, using
appropriate contact time. If a spill occurs, the first step should be to
• Rinse the spill site with water.
• Dispose off all materials in appropriate biohazard containers. • assist/evacuate personnel, and then
• confinement and cleanup of the spill can begin.
Bloodborne Pathogens
RADIATION SAFETY
• Special precautions must be taken when handling all specimens because of the
continual increase in the proportion of infectious samples received in the laboratory. Environmental Protection

Airborne Pathogens • 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
• Those workers in high-risk areas may be required to wear a respirator for personnel only.
protection
• Protective measures in the clinical laboratory generally involve work practice Personal Protection
and engineering controls focused on prevention of aerosolization,
containment/isolation, and respiratory protection of N-95 (filtration of 95% of • Users should be monitored to ensure that the maximal permissible dose of
particles >0.3 μm) or better. radiation is not exceeded.
• Radiation monitors must be evaluated regularly to detect degree of exposure
Shipping for the laboratory employee.
• Records must be maintained for the length of employment plus 30 years.
• Known or suspected infectious specimens are labeled infectious substances if
the pathogen can be readily transmitted to humans or animals. Non-ionizing Radiation
• Diagnostic specimens are those tested as routine screening or for initial
diagnosis. Each type of specimen has rules and packaging requirements. • These energies have varying biologic effects, depending on wavelength, power
intensity, and duration of exposure. Laboratorians must be knowledgeable
• Arrangements for the storage of chemicals will depend on the quantities of regarding the hazards presented by their equipment to protect themselves and
chemicals ancillary personnel.
• Proper storage is essential to prevent and control laboratory fires and accidents

CHEMICAL SAFETY

Storage and Handling of Chemicals

• Arrangements for the storage of chemicals will depend on the quantities of

chemicals needed and the nature or type of chemicals.
• Proper storage is essential to prevent and control laboratory fires and
accidents

FIRE SAFETY

Classification of Fires

• Class A: ordinary combustible solid materials, such as paper, wood, plastic, and
fabric
• Class B: flammable liquids/gases and combustible petroleum products○
• Class C: energized electrical equipment
• class D: combustible/reactive metals, such as magnesium, sodium, and
potassium
Flammable/Combustible Chemicals Types and Applications of Fire Extinguishers
• They are classified according to flash point, which is the temperature at which
sufficient vapor is given off to form an ignitable mixture with air.
• Some commonly used flammable and combustible solvents are acetone,
benzene, ethanol, heptane, isopropanol, methanol, toluene, and xylene.
• It is important to remember that flammable or combustible chemicals also
include certain gases, such as hydrogen, and solids, such as paraffin.

Corrosive Chemicals

• Typical examples include acids (acetic, sulfuric, nitric, and hydrochloric) and
bases (ammonium hydroxide, potassium hydroxide, and sodium hydroxide).
• External exposures to concentrated corrosives can cause severe burns and
require immediate flushing with copious amounts of clean water

Reactive Chemicals

• Some strong acids or bases react with water to generate heat. PSC
• Hydrogen is liberated if alkali metals (sodium or potassium) are mixed with
water or acids, and spontaneous combustion also may occur.
• The mixture of oxidizing agents, such as peroxides, and reducing agents, such
as hydrogen, generates heat and may be explosive.
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CONTROL OF OTHER HAZARDS Ergonomic Hazards

Electrical Hazards • The primary contributing factors associated with repetitive strain disorders are
position/posture, applied force, and frequency of repetition.
• precautionary procedures to follow when operating or working around • Remember to consider the design of hand tools (e.g., ergonomic pipettes),
electrical adherence to ergonomically correct technique, and equipment positioning
• Use only explosion-rated (intrinsically wired) equipment in hazardous when engaging in any repetitive task
atmospheres.
• Be particularly careful when operating high-voltage equipment, such as DISPOSAL OF HAZARDOUS MATERIALS
electrophoresis
• Use only properly grounded equipment (three-prong plug). Chemical Waste
• Check for frayed electrical cords.
• flush water-soluble substances down the drain with copious quantities of
water
• Promptly report any malfunctions or equipment producing a “tingle” for repair.
• Do not work on “live” electrical equipment. Radioactive Waste
• Never operate electrical equipment with wet hands.
• Know the exact location of the electrical control panel for the electricity to • - Many clinical laboratories transfer radioactive materials to a licensed receiver
your work area. for disposal.
• Use only approved extension cords in temporary applications and do not
overload circuits. (Some local regulations prohibit the use of any extension Biohazardous Waste
cord
• - All biomedical waste should be placed in a bag marked with the biohazard
• Have ground, polarity, and leakage checks and other periodic preventive
symbol and then placed into a leakproof container that is puncture resistant
maintenance performed on outlets and equipment.
and equipped with a solid, tight fitting lid. All containers must be clearly
Compressed Gas Hazards marked with the word biohazard or its symbol.

• Know the gas that you will use. DISPOSAL OF HAZARDOUS MATERIALS
• Store tanks in a vertical position.
• All sharp instruments, such as needles, blades, and glass objects, should be
• Keep cylinders secured at all times.
placed into special puncture-resistant containers before placing them inside
• Never store flammable liquids and compressed gases in the same area.
the bag and container.
• Use the proper regulator, tubing, and fittings for the type of gas in use.
• Needles should not be transported, recapped, bent, or broken by hand.
• 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.
III. PREPARATION OF SOLUTIONS AND DILUTIONS
• Make certain that acetylene tanks are properly piped (the gas is incompatible
with copper tubing) REPORTING OF MEASUREMENTS

• Do not force a “frozen” or stuck cylinder valve. Components of a Laboratory Result:

• Use a hand truck to transport large cylinders.
• Always check cylinders on receipt and then periodically for any problems such • Actual value
as leaks. • Unit
• Make certain that the cylinder is properly labeled to identify the contents.
• Empty tanks should be marked “empty. ➢ It is recommended that analytes be reported using moles of solute per volume of
solution (substance concentration)
Cryogenic Materials Hazards ➢ Reporting laboratory results is often expressed in terms of substance concentration
(e.g. moles) or the mass of a substance (e.g. mg/dL, g/L, mEq/L and IU) rather than
• Only containers constructed of materials designed to withstand ultralow SI units
temperatures should be used for cryogenic work.
• In addition to the use of eye/face protection, hand protection to guard against SOLUTIONS
the hazards of touching supercooled surfaces is recommended.
• The gloves, of impermeable material, should fit loosely so that they can be Solvent + Solute= Solution
taken off quickly if liquid spills on or into them.
SOLUTION
• Also, to minimize violent boiling/frothing and splashing, specimens to be
frozen should always be inserted into the coolant very slowly. • A homogenous mixture of two or more substances with each substance
• Cryogenic fluids should be stored in well-insulated but loosely stoppered retaining its own chemical identity
containers that minimize loss of fluid resulting from evaporation by boil-off • A solution contains two or more components: a solvent and one or more
and that prevent plugging and pressure buildup solutes.
• Solutions used in laboratories and clinical settings are most often liquids, and
Mechanical Hazards
the solvent is nearly always water.
Centrifuges
SOLVENT SOLUTE
• must be balanced to distribute the load equally. It is the component of a solution that is
It is the component of a solution that is
• The operator should never open the lid until the rotor has come to a complete present in the greatest amount
present in a lesser amount relative to that
stop. of the solvent
• Safety interlocks on equipment should never be rendered inoperable. TYPES OF SOLUTIONS

Laboratory glassware • Colloidal solution

• Emulsions
• Glass beads or boiling chips, should be added to help eliminate bumping/boil • Saturated solution PSC
over when liquids are heated. Tongs or insulated gloves should be used to • Unsaturated solution
remove hot glassware from ovens, hot plates, or water baths. Glass pipettes
should be handled with extra care, as should sharp instruments such as cork COLLOIDAL SOLUTION
borers, needles, scalpel blades, and other tools.
• Foam
• Milk
• Smoke
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• Detergents 2. What is the percent-by-volume concentration if a 2 mL of concentrated HCl is diluted

• Gel with 80 mL distilled water?
• Blood • Given:
• Paint ▪ 2mL conc. HCl
• Cosmetics ▪ 80mL dH2O
• Solution:
EMULSIONS Percent by volume = volume of solute x100
volume of solution
SATURATED SOLUTION
= 2mL conc.HCl x100
2mL conc. HCl + 80mL dH2O
= 2 mL conc. HCl x100
82 mL solution
= 2.44% HCl solution

3. What is the concentration of a 200 mL solution containing 1.8g of NaCl?

• Given:
▪ 1.8g NaCl
▪ 200mL solution
• Solution:
Mass-volume percent = mass of solute x100
volume of solution
= 1.8g NaCl x100
UNSATURATED SATURATED 200mL solution
More solute dissolves No more solute dissolves = 0.9% NaCl solution
CONCENTRATION OF SOLUTIONS = 0.9g/mL NaCl solution

• refers to the weight or volume of the solute present in a specified amount of CALCULATING THE AMOUNT OF SOLUTE OR SOLVENT IN A GIVEN PERCENT SOLUTION
solvent or a solution
1. Normal saline solution (NSS) is used to dissolve drugs for IV use which is 0.9% w/v
THREE BASIC TYPES OF SOLUTIONS: NaCl in water. How many grams of NaCl is needed to prepare a 50mL NSS?
• Given:
• Percent solutions
▪ Volume of solution = 50mL
• Molar solutions
▪ Mass-volume percent = 0.9% NaCl solution (NSS)
• Normal solutions
• Solution:
Mass-volume percent = mass of solute x100
1. PERCENT SOLUTIONS
volume of solution
• Amount of solute in a solution can be measured as a percentage of the total
volume of the solution 0.9% = x x100
• Expressed as equal parts per hundred or the amount of solute per 100 total 50mL
units of solution 0.9%(50mL) = x(100)
• Three expressions of percent solution: 0.9%(50mL) = x(100)
100 100
Percent by volume
Percent by mass (mass- Mass-volume percent x = 0.45g NaCl
(volume- volume percent
mass percent or %w/w) (%w/v)
or %v/v) 2. 10% bleach (Sodium hypochlorite) is used to disinfect benches before and after
work. It denatures protein in micro-organisms and is therefore effective in killing
is the mass of solute in a bacteria, fungus and viruses. How much bleach is needed to make 100mL of 10%
is the mass of solute in a
is the volume of solute in a bleach (sodium hypochlorite) solution? How much distilled water is needed to dilute
solution divided by the solution (in grams) divided
solution divided by the the bleach?
total mass of solution, by the total volume of
total volume of solution,
solution (in milliliters), • Given:
multiplied by 100 (to put
multiplied by 100. ▪ Volume of solution = 100mL
the value in terms of multiplied by 100.
▪ Percent by volume = 10% bleach (sodium hypochlorite)
percentage).
• Solution:
Percent by volume = volume of solute x100
mass of solute x100 volume of solute x100 mass of solute (g) x 100 volume of solution
mass of solution volume of solution volume of solution (mL)
10% = x x100
Practice Question: 100mL
10%(100mL) = x(100)
1. What is the percent-by-mass concentration of sucrose in a solution made by 10%(100mL)= x(100)
dissolving 7.5g of sucrose in 86.5g of water? 100 100
• Given:
▪ 7.5gsucrose x = 10mL bleach (sodium hypochlorite)
▪ 86.5g water
• Solution: 2. MOLAR SOLUTIONS
Percent by mass = mass of solute x 100 • Solution containing one gram molecular weight (one mole of the solute in one
mass of solution liter solution) of the substance per liter of the solution

= 7.5g sucrose x100 M = g of solute

7.5g sucrose + 86.5g water MW x L of solution
= 7.5 g sucrose x100
94 g solution M = mol of solute
= 7.98% sucrose solution L of solution
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Practice Question: 1. Determine the normality of a solution containing 15g KCl dissolved in enough
water togive 0.20L solution. (K-39.10; Cl-35.45)
1. Determine the molarity of a solution containing 4.35 moles of KMnO4 dissolved in
enough water to give 750 mL solution
• Given:
▪ 4.35moles KMnO4
▪ 750mL solution Conversion:
• Solution: 750mL x 1L = 0.75L
M = mol of solute
1000mL
L of solution

= 4.35 moles
KMnO4 0.75L
= 5.8M KMnO4 = 1.01N KCl solution

2. Determine the molarity of a solution containing 20g NaOH dissolved in enough Relationship of Normality and Molarity
water to give 1.50L solution. (Na-23; O-16, H-1)
• Normality is ALWAYS equal or greater than molarity of that compound.
• Given:
▪ 20g NaOH Practice Questions:
Molecular weight:
▪ 1.50L solution
• Solution: NaOH = 23 + 16 + 1 a) What is the molarity of a 2N NaCl solution?
M = g of solute = 40g/mol b) What is the normality of a 5M H2SO4 solution?
MW x L of soln’
Solutions:
= 20g NaOH
40g/mol x 1.50L a) M = Normality/valence
= 0.33M NaOH solution = 2/1
= 2M NaCl solution
a) N = Molarity x valence
3. How many grams of FeSO4 is needed to prepare 0.3L of 0.10M FeSO4 solution? (Fe- =5x2
55.85;S-32.06; O-14) = 10N H2SO4 solution
• Given: Molecular weight:
▪ 0.3L solution DILUTIONS
FeSO4 = 55.85 + 32.06 + 16(4)
▪ 0.10M
• Solution: • represents the ratio of concentrated or stock material to the total final volume
= 55.85 + 32.06 + 64 of a solution and consists of the volume or weight of the concentrate plus the
M = g of solute
MW x L of soln’ volume of the diluent, with the concentration units remaining the same.
= 151.88g/mol
• In the molar, normal or percentage solutions, the amount of solute contained
0.10M = x _ in a given volume of solution is equal to the product of volume times the
151.88g/mol x 0.30L concentration.
x = 0.10M (151.88g x mol x 0.30L) • Whenever the solution is diluted, the volume is increased and its
x = 4.56g FeSo4 concentration is decreased but the total amount of solute remains
unchanged.

3. NORMAL SOLUTIONS TYPES DILUTIONS

• Least likely to be encountered of the three concentration expressions to
be encountered in the clinical laboratories, but is often used in chemical − Simple Dilution
titrations and chemical reagent classification − Serial Dilution
• The number of gram equivalent weight per 1 L of solution.
SIMPLE DILUTION
N = g of solute
EW x L of solution Equivalent Weight = MW / valence

IDENTIFYING THE VALENCE OF ACIDS, BASES, AND SALTS

• ACIDS – count the number of Hydrogen ions

• BASES – count the number of Hydroxide ions
• SALTS – multiply the absolute value of the ions

ACID BASE SALT

HCl – 1 NaOH – 1 NaCl – Na(+1) x Cl(-1) = 1
HNO3-1 KOH – 1 CaCl2 - Ca (+2) x Cl(-1) = 2
H2SO4 –2 NH4OH– 1 FeCl3 – Fe (+3) x Cl(-1) = 3
H3PO4 -3 Ba(OH)2– 2

Ex. Find the GEW of KOH.

Solution. Determine the GMW of the compound KOH

K - 39.1 56.1 g
O - 16.0 1
H – 1.0 . = 56.1 g/Eq
56.1g/mol
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SERIAL DILUTION • CENTRIFUGE

− Usually, blood chemistry large amount of sample is needed for chemistry
analysis
− Ex. Lipid profile
− When doing venipuncture, you extract blood in the median, cephalic or basilic
at the antecubital fossa
− For geriatric and pediatric patient, we extract blood from the dorsal surface of
the hand or foot
− 3-4 inches from the bend of the elbow
− Median-cephalic-basilic vein

COLLECTION PROCEDURE

- Disinfection (hands, working station)

- Check the request form of the patient
o Proactively ask the identity of the px (name, b-day, age)
o Ask for ID
o Compare to the request form
Preparation of A Working Solution from A Stock Solution - Prepare and inspect the materials
o Check the needle if properly attached and check the plunger for air
C1V1=C2V2
bubbles (NO AIR!!!)
C1 = concentration of stock solution - Properly apply the torniquet
V1 = volume of stock solution o Ask the px if has allergies
C2 = concentration of the working solution o Apply 2 inches from the bend of elbow
V2 = volume of the working solution - Inspect the site of puncture
- Ask the px to close the fist
- Select the vein
Practice Question: ANTECUBITAL FOSSA
- BASILIC- near towards the body and nerve-endings (branchial
1. What is the initial volume of a 40% formaldehyde diluted to prepare 100mL of artery)
10%formaldehyde solution? - CEPHALIC- second choice, away from body
• Given: - MEDIAN CUBITAL- middle, best site for venipuncture
V1 = ? C1 = 40% V2 = 100mL C2 = 10% - Apply the anti-septic (circular motion or inner to outside)
• Solution: - Dried first before puncture
- Hold the syringe properly (15-30⁰ angle)
C1V1 = C2V2
- Insert the needle, BEVEL UP!
(40%)V1 = (10%)(100mL) - Remove the torniquet before the needle
(40%)V1 = (10%)(100mL) - Apply cotton after the needle, PATIENT CARE!!
40% - Remove the needle, insert the spx sideways
o EDTA- purple/ lavender, 8-10 inversions, CBC
V1 = 1000ml o Red- glass-no inversion, plastic- 5x
40 - Label the tube
V1 = 25mL o Name of px
o Date and time
2. What is the final concentration of a 50mL 90% methanol diluted to prepare a o Age/ Sex
200mLmethanol solution? o Initial of phleb
• Given: - DISNIFECT
▪ V1 = 50mL
▪ V2 = 200mL POST-LAB
• Solution:
COLLECTION AND HANDLING OF LABORATORY SPECIMEN
C1V1 = C2V2
Describe the appearance of the following processed specimen:
(90%)50mL = C2(200mL)
(90%)50mL = C2(200mL) a. Jaundice or Icteric Serum or plasma
200mL 200mL b. Lipemic blood, serum or plasma
c. Hemolyzed serum
C2 = 2000
200
C2 = 10% methanol solution

IV. COLLECTION AND HANDLING OF LABORATORY SPECIMENS

PRE-LAB

MATERIALS

• TOURNIQUET
• STERILE DRY SYRING
• STREILE NEEDLES (GAUGE 21)
• COTTON OR GAUZE PAD
• 70% ALCOHOL
• ANTICOAGULATED TUBES
• PLAIN TUBES
• APPLICATOR STICKS
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AST – ASPARTATE AMINOTRANSFERASE

ALT – ALANINE AMINOTRANSFERASE
CK – CREATINE KINASE
ALP – ALKALINE PHOSPHATSE
ACP – ACID PHOSPHATASE
PO4 – PHOSPHATE
LD– LACTATE DEHYDROGENASE

Why is that centrifugation of some

What can cause a blood specimen to become hemolyzed?
blood specimen is necessary?

− Centrifugation is a process in
which centrifugal force is
used to separate solid matter
from a liquid suspension.

Why is that centrifugation of some

blood specimen is necessary?

− Most clinical chemistry tests

require a sample of serum or plasma

- Tests performed in serum or plasma

- Mention some changes that may occur whenever serum is not removed from clot after
collection

- What blood analytes are affected by food intake?

- What substances are increased in a hemolyzed substances?

ENUMERATE WAYS TO AVOID HEMOLYSIS

• Use the correct needle size for blood collection (20-22 gauge).
• Avoid using butterfly needles, unless specifically requested by patient.
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• Warm up the venipuncture site to increase blood flow.

• Allow disinfectant on venipuncture site to dry completely.
• Collect blood sample in the correct blood collection tube
• Collect the correct volume for the tube size. Use smaller tubes for difficult
draws.
• Coagulation: place the tube in an upright position for 15-30 minutes at room
temperature. Then store at 4°C.
• Centrifuge samples for serum separation within 4 hours of sample collection.
• If tests are performed within 48 hours of collection, transfer serum to a sample
tube (cryovial) and store at 4°C.
• If tests will be performed more than 48 hours after collection, aliquot samples
and store at -20°C or lower.