CLINICAL CHEMISTRY

[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

OUTLINE
o understands fundamental concepts critical to any
BASIC PRINCIPLES AND PRACTICES analytic procedure
I. UNITS OF MEASURE • Must understand the tools:
1. SI units o Equipment – checks if it functions well
SI conversions o Reagent – check the integrity in terms of supply,
II. REAGENTS expiration date
1. Chemicals o Principle of the testing methods
2. Reference materials
o Knowledge of the medical uses of the
3. Water specifications
Classifications of water determinations
4. Solution properties
Concentration UNITS OF MEASURE
Colligative properties • Quantitative measurement is expressed in defined units
Redox potential
o Number – related to the actual test value
Conductivity
pH and buffers o Unit – defines the physical quantity or dimension:
III. CLINICAL LABORATORY SUPPLIES mass, length, time, or volume
1. Thermometers ▪ Substance concentration units: moles
Classifications of thermometers ▪ Derived units: mg/dL, g/dL, mEq/L, IU
2. Glassware and plasticware a. Liter – reference unit of volume
Glassware
Plasticware
Classifications SI UNITS
3. Desiccators and desiccants • International System of Units
4. Balances • Based on the Metric System
IV. BASIC CENTRIFUGATION TECHNIQUES • Systeme International d’unites (French)
1. Centrifugation
2. Filtration
3. Dialysis Table No. 1 SI units
Base quantity Name Symbol
Length Meter m
● CLINICAL CHEMISTRY Mass Kilogram kg
o Basic science that utilizes the specialty of Time Second s
chemistry to study human beings
Electric current Ampere A
o Applied science when analyses are performed on
Thermodynamic Kelvin K
body fluids or tissues for diagnosis or treatment of
temperature
disease
Amount of substance Mole mol
• MALFUNCTION
o Trauma or by invasive agents – parasites, Luminous intensity Candela cd
bacteria, viruses
o Genetic deficiency of a vital enzyme Table No. 2 Prefixes used with SI units
o Defect in the cellular recognition of signals Factor Prefix Symbol Select decimals
o Insufficient supply of blood, nutrients, and oxygen 10-18 Atto a -
o Malignancy – tumors that can cause cancer 10-15 Femto f -
10-12 Pico p -
ROLE OF CLINICAL CHEMISTRY LABORATORY 10-9 Nano n -
• Perform analytic procedures that yield accurate and 10-6 Micro µ 0.000001
precise information, aiding in patient diagnosis and 10-3 Milli m 0.001
treatment 10-2 Centi c 0.01
• Measure chemical changes in the body for diagnosis, 10-1 Deci d 0.1
therapy, and prognosis of disease
10-0 Liter, meter, gram Unit 1.0
o Major Sample: Blood (specifically, plasma/serum)
101 Deka da 10.0
• Measure the concentration of a particular constituent (the
analyte) in body fluids 102 Hecto h 100.0
o Analytes: Creatinine, Blood urea nitrogen, uric 103 Kilo k 1000.0
acid, blood glucose, cholesterol 106 Mega M 1000000.0
o Measurement of electrolytes such as K, Na, & Ca 109 Giga G -
1012 Tera T -
1015 Peta P -
ROLES OF MEDICAL TECHNOLOGIST 1018 Exa E -
• Achievement of reliable results:
o must be able to correctly use basic supplies and
equipment

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 1
 CLINICAL CHEMISTRY
[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

WATER SPECIFICATIONS
SI CONVERSIONS • Distilled water
• To convert between SI units, move the decimal by the o Purified to remove almost all organic materials
difference between the exponents represented by the through distillation where water is boiled and
prefix: vaporized
o to the right (from a larger to a smaller unit) o Liquid state is collected
o to the left (from a smaller to a larger one) o Tap water is not allowed
0.0 • Deionized water
decimal point o Purified by ion exchange
o Water is produced using either an anion or cation
REAGENTS exchange resin and is followed by replace of the
• Ready-to-use (so MTs will only need to put water or buffer) removed ions with hydroxyl or hydrogen ions
o Removes some or all ions and is purified
• Types:
previously by treated water (predistilled/distilled)
o Commercially prepared reagents
o Excellent in removing dissolved ionized solids
▪ Usually provided by the suppliers of
and dissolved gases
automated machines
o In-house prepared reagents • Reverse Osmosis (RO) water
▪ Requires preparation mixing of reagents o Uses pressure to force water through a
semipermeable membrane
CHEMICALS o Can be used for the pretreatment of
• Analytical Grade (AR) water and does not remove dissolved
o Suitable for most analytical procedures gases
o Carry designations as “AR” (analytical reagent) or • Ultrafiltration and Nanofiltered Water
“ACS” (American Chemical Society) and “For o Excellent in removing particulate matters,
Laboratory Use” or “ACS Standard-Grade microorganisms, and any pyrogens or endotoxins
Reference Materials” o Through UV oxidation which removes trace
• Ultrapure Chemicals organic materials; sterilization process at specific
o Suitable for techniques that require extremely wavelengths; and combined with ozone treatment
pure chemicals (e.g., AAS, EIA, MDx) (in the form of gas) that destroy bacteria.
▪ Been through additional purification • Reagent Grade Water
steps for use in specific procedures such o Obtained by initial filter, followed by RO,
as chromatography, atomic absorption, deionization and a 0.2 mm filter
immunoassays, molecular diagnostics, o Production of this water depends on the condition
etc. of the feed water
▪ Carry designations: “HPLC” (High
performance liquid chromatography) or CLASSIFICATIONS OF WATER
“Chromatographic” • Type I Water
• Chemically Pure (CP) o Used for test methods that requires minimum
o Impurity limitations are not stated interference such as trace metal analysis by
o Preparation is not uniform Frame Emission Spectrometry (FES) and Atomic
o Not recommended for clinical labs Absorption Spectrometry (AAS)
• United States Pharmacopeia (USP) & National o Also used for blood gas, pH, enzyme and
Formulary (NF) Grade electrolyte analysis
o Used to manufacture drugs • Type II Water
o Standards are based on the criterion of not being o For analytical preparations
injurious to man o Also used in reagent, QC, and standard
• Technical or Commercial Grade preparation
o Used for manufacturing • Type III Water
o Should not be used in clinical labs o Used for glassware washing but not for analysis
and reagent preparations
REFERENCE MATERIALS
• Primary Standard SOLUTION PROPERTIES
o Highly purified chemical that can be measured • Solute
directly to produce a substance of exact known o Substance dissolved in a liquid
concentration and purity o In laboratory science, these biologic solutes are
• Secondary Standard known as analytes
o Substance of lower purity with concentration • Solvent
determined by comparison with a primary o Liquid which the solute is dissolved
standard

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 2
 CLINICAL CHEMISTRY
[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

CONCENTRATION o Reciprocal of conductivity

Routinely, concentration is expressed as percent solution, molarity, o Measure of a substance resistance to the
molality, or normality concentration is expressed as percent passage of electrical current
solution, molarity, molality, or normality.
• Percent solution pH AND BUFFERS
o Expressed as equal parts per hundred or the • Buffers
amount of solute per 100 total units of solution o weak acids or bases and their related salts
▪ weight per weight (w/w) ▪ dissociation characteristics minimize
▪ volume per volume (v/v) changes in the hydrogen ion
• it is recommended to use concentration
grams per deciliter (g/dL) • pH
▪ weight per volume (w/v) o represents the negative/inverse log of the
• Molarity (M) hydrogen ion concentration
o Number of moles per 1 L of solution o ranges from 0 to 14
• Molality (m)
o Amount of solutes per 1 kg of solvent Equation No. 1 pH
• Normality
o Often used in chemical titrations and chemical
reagent classification
o Number of gram equivalent weights per 1 L of
solution
Where [H+] = the concentration of hydrogen ions in mol/L
COLLIGATIVE PROPERTIES
The behavior of particles or solutes in solution demonstrates four • Henderson-Hasselbalch equation
repeatable properties based only on the relative number of each o Describes the dissociation characteristics of
kind of molecule present weak acids (pKa) and bases (pKb) and the effect
on pH:
• Osmotic pressure
o Pressure that opposes osmosis when a solvent
flows through a semipermeable membrane to Equation No. 2 Henderson-Hasselbalch equation
establish equilibrium between compartments of
differing concentration
o Osmotic pressure is proportional to the
When the ratio of [A-] to [HA] is 1, pH = pK and the buffer has its greatest
concentration of molecules in a solution buffering capacity
• Vapor pressure
o Pressure at which the liquid solvent is in • Ionic strength
equilibrium with the water vapor o Concentration or activity of ions in a
• Freezing point solution/buffer
o Temperature at which the vapor pressures of the
solid and liquid phases are the same Equation No. 3 Ionic strength
• Boiling point
o Temperature at which the vapor pressure of the
solvent reaches one atmosphere

REDOX POTENTIAL
Where Ci = concentration of the ion
• AKA oxidation-reduction potential Zi = charge of the ion
• Measure of the ability of a solution to accept or donate ∑ = sum of the quantity (Ci)(Zi)2 for each ion present
electrons
• Reducing agents CLINICAL LABORATORY SUPPLIES
o Substances that donate electrons THERMOMETERS
• Oxidizing agents
• Analytical reactions occur at an optimal temperature
o Substances that accept electrons
• Usage of circulating water or heating/cooling metal blocks
• Integral/need to be placed
LEO the lion says GER
(lose electrons oxidized) (gain electrons reduced)
CLASSIFICATIONS OF THERMOMETERS
• Liquid-in-glass
CONDUCTIVITY
o Colored liquid or mercury encased in plastic/glass
• Measure of how well electricity passes through a solution material with a bulb at one end a graduated stem
• Conductivity (ohms-1) quality depends on the number of o Usually measures between 20°C-400°C
respective charges of ions present in the solution o Types:
• Resistivity (ohms) o

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 3
 CLINICAL CHEMISTRY
[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

▪ Total Immersion o Begins to replace glassware in the lab setting due

• The entire thermometer (bulb to high resistance to corrosion, breakage,
and liquid column) is exposed disposable and flexibility.
to the temperature being A. Polystyrene
measured B. Polyethylene
• Most commonly used in C. Polypropylene
refrigerator, freezer, incubator - D. Tygon
Should be placed horizontally E. Teflon
for the temperature to be read F. Polycarbonate
G. Polyvinyl chloride

CLASSIFICATIONS OF GLASS/PLASTICWARES
• Laboratory Vessels – designed to deliver (TD) or to
▪ Partial Immersion contain (TC)
• Has an immersion line and o Volumetric flask
should be immersed to the ▪ Calibrated to hold one exact volume of
proper height liquid.
• Used in water/cold baths o Erlenmeyer flask & Griffin Beaker
• Should be used vertically ▪ Hold different volume
▪ Used in reagent preparation
▪ Flask is usually used in microbiology
Immersion line – the part to be submerged for section for preparation of agar plate
the temp to be read o Graduated Cylinder
• Surface Thermometer - For ▪ Used to measure volumes of liquid
checking temperature on flat • Pipets – Glass or plastic utensils used to transfer liquid
surfaces such as oven and o Design:
incubator ▪ To Contain
• Electronic Thermometer • Does not dispose exact
o Thermistor Probe amount
o Fast reading, millisecond response time • Used for viscous samples
• Digital Thermometer • Uses mercury as calibrating
o Used in body temperature medium
• Proper use requires rinsing
GLASSWARE AND PLASTICWARE technique
• Commonly used in laboratory set-ups such as test tubes, • Hold Volume but does not
pipettes, etc. dispense exact volume
▪ To Deliver
• Used for non-viscous samples
GLASSWARE
• Uses distilled water as
• Borosilicate
calibrating medium
o Kimax ® / Pyrex ®
• Dispense the exact volume
o Used for heating and sterilization
o Calibration marks/ drainage characteristics:
o Most commonly used
▪ Blowout Pipette
• Aluminosilicate
• Used in Serologic/Oswald-
o Corex ®
Folin measuring
o Strengthened chemically than thermally
o 6x stronger than borosilicate • With etched ring/two small
continuous rings
• High silica
o Boron •
o High alkali resistant Figure No. 1 Blowout pipet
• Acid / Alkali resistant (Vycor)
o Utilized for high thermal blastic shock and
extreme chemical treatment with acids (except
hydrofluoric acid).
• Amber Colored (Low actinic)
o Protect photosensitive reagents 2 rings; Note that when a substance is still present, removing by
• Lime Soda (Flint glass) blowing is necessary
o Made-up of soda lime glass and mixture of Ca, Si, ▪ Self-Draining
and sodium oxides. • Used in Volumetric/Mohr
o Poor resistance to high temperatures. measuring
• Without marking
PLASTICWARE • Drains completely

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 4
 CLINICAL CHEMISTRY
[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

Figure No. 2 Self-draining pipet Figure No. 4 Correct and incorrect pipet positions

o Types (According to Use)

▪ Measuring or Graduated Pipets
• Graduated uniformly along its
length Figure No. 5 Correct pipet operation
• Designed to deliver any
amount within its capacity
• Serologic Pipet: has
graduation marks to the tip
• Examples:
o Blowout Pipet:
continuous etched
ring
o Mohr Pipet: no
graduation marks to
the tip
Figure No. 3 Serological vs Mohr pipets

Table No. 4 Volumetric vs Ostwald-Folin pipets

Volumetric pipet Ostwald-Folin pipet
Size Larger Smaller
Location Located at the center Located closer to the
of the bulb (symmetrical) delivery tip
▪ Volumetric or Transfer Pipette Design To deliver To deliver
• Delivers an exact volume
• Examples: Figure No. 6 Serological, Mohr, Volumetric, & Ostwald-Folin pipets
o Ostwald-Folin Pipet:
for viscous fluids
(blowout pipet)
o Volumetric Pipet: for
aqueous solutions
(self-draining)
Table No. 3 Pipet classification
Pipet classification
Design
To contain (TC) to deliver (TD)
Calibration marks/drainage characteristics o Mechanical and Automatic Pipets
Blowout Self-draining ▪ Micropipette – deliver amount > 1mL
Type ▪ Micropipette – deliver amount < 1mL
Measuring or graduated Transfer ▪ Dilutor or Dispenser
• Serologic • Volumetric • Automatic pipets that obtain
• Mohr • Ostwald-Folin the liquid from a common
• Bacteriologic • Pasteur pipets reservoir and dispense it
• Ball, Kolmer, Kahn • Automatic repeatedly
• Micropipette macropipettes/ • The dispensing pipets may be
micropipettes bottle-top, motorized,

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 5
 CLINICAL CHEMISTRY
[TRANS] UNIT 1: BASIC PRINCIPLES AND PRACTICE

handheld, or attached to a o Depends on

dilutor. ▪ Mass
▪ Burets ▪ Speed
• For dispensing liquid during ▪ Radius
titration o Expressed by:
▪ Syringes ▪ Revolution per minute (RPM) (speed)
• Used to transfer small volumes ▪ Relative centrifugal force (RCF) or
in blood gas analysis, gravities (g) (centrifugal force)
chromatography, or
electrophoresis Equation No. 4 Relative centrifugal force

DESICCATORS AND DESICCANTS Where 1.118x10-5 is a constant determined from the angular velocity;
• Desiccators r = radius in cm, measured from the center of the centrifuge axis to the bottom of test
o Closed and sealed containers that contain desiccant tube
material
o Uses hygroscopic (substances that take up water • Centrifuge must be properly balances and free from
when exposed to atmospheric conditions) substance excessive vibrations. If only 1 tube needs to be centrifuged,
that take up water/moisture on exposure to air make a balancer where it has the same amount of volume
o Hydrate as the other tube.
▪ Compound and the associated water
o Anhydrous Figure No. 8 Properly balanced centrifuge
▪ when water of crystallization is removed from
the compound
• Desiccants
o Drying agents
o Materials that remove moisture from the air and other
materials

Figure No. 7 Desiccator with desiccant

FILTRATION
• Alternative method to separate solid from liquid
• Filter materials: paper, cellulose, polyester fibers, and
column materials.
• Filtrate
• Deliquescent substances o Liquid that passes through a filter
o Compounds that absorb enough water from the
atmosphere to cause dissolution DIALYSIS
• Method for separating macromolecules from a solvent of
BALANCES smaller substances
• Top-loading Balance / Electronic Top-Loading Balance o A solution is put into a bag or is contained on one
o For knowing the mass of substances (greater quantity) side of a semipermeable membrane
o Used for preparative experiments ▪ Larger molecules are retained within the
• Analytical Balance sack or on one side of the membrane
o For preparation of primary standards ▪ Smaller molecules and solvents diffuse
o Measure exact mass but with lower capacities out
(Operating ranges 0.01 mg to 160 g) • Popularized by the Technicon AutoAnalyzer system
SUBSUB TOPIC FORMAT

BASIC SEPARATION TECHNIQUE

CENTRIFUGATION
• A process in which a centrifugal force is used to separate
solid matter from a liquid suspension
• Consists of head/rotor (attached to the shaft of the motor)
carrier and shields.
• The centrifugal force:

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 6