CLINICAL CHEMISTRY

SMTY1209: CLINICAL CHEMISTRY [LEC]

OUTLINE
o Analytes: Creatinine, Blood urea nitrogen, uric
BASIC PRINCIPLES AND PRACTICES acid, blood glucose, cholesterol
o Measurement of electrolytes such as K, Na, & Ca
I. UNITS OF MEASURE/REAGENTS
1. Chemicals
2. Reference Materials ROLES OF MEDICAL TECHNOLOGIST
3. Water Specifications  Must understand the tools:
4. Solution Properties o Equipment – checks if it functions well
5. Concentration o Reagent – check the integrity in terms of supply,
6. Colligative Properties expiration date
7. Redox Potential o Principle of the testing methods
8. Conductivity o Knowledge of the medical uses of the
9. Ph and Buffers determinations
II. CLINICAL LABORATORY SUPPLIES  Units of Measure
1. Thermometers/Temperature o Primary purpose of a clinical chemistry laboratory:
2. Glassware and Plasticware to perform analytic procedures that yield accurate
3. Desiccators and Desiccants and precise information, aiding in patient
4. Balances diagnosis and treatment
III. CENTRIFUGATION o Achievement of reliable results: clinical laboratory
IV. LABORATORY MATHEMATICS & scientist must be able to correctly use basic
CALCULATIONS supplies and equipment and possess and
1. Significant Figures understanding of fundamental concepts critical to
2. Logarithms any analytic procedure
3. Concentrations o Quantitative measurement is expressed in
4. Dilutions defined units
5. Water of Hydration o Components of quantitative laboratory results:
6. Graphing and Beer’s Law  Number – related to the actual test
V. SPECIMEN CONSIDERATIONS value
1. Types of Samples  Unit – defines the physical quantity or
2. Sample Processing dimension: mass, length, time, or
3. Sample Variables volume
4. Chain of Custody  Substance concentration units:
5. Electronic and Paper Reporting of moles
Results  Derived units: mg/dL, g/dL,
mEq/L, IU
a. Liter – reference unit of volume
 SI Units
BASIC PRINCIPLES AND PRACTICES o International System of Units
UNITS OF MEAURE o Based on the Metric System
● CLINICAL CHEMISTRY o Systeme International d’unites (French)
o Basic science that utilizes the specialty of
chemistry to study human beings Base quantity
o Applied science when analysis are performed on Meter (m) Length
body fluids or tissues for diagnosis or treatment of Kilogram (kg) Mass
disease Second (s) Time
 MALFUNCTION Mole (mol) Quantity of
o Trauma or by invasive agents – parasites, substance
bacteria, viruses Ampere (A) Electric current
o Genetic deficiency of a vital enzyme Kelvin (K) Thermodynamic
o Defect in the cellular recognition of signals temperature
o Insufficient supply of blood, nutrients, and oxygen
Candela (cd) Luminous intensity
o Malignancy – tumors that can cause cancer
REAGENTS
ROLE OF CLINICAL CHEMISTRY LABORATORY
o Ready-to-use (so MTs will only need to put water
 Measure chemical changes in the body for diagnosis, or buffer)
therapy, and prognosis of disease o Types:
o Major Sample: Blood (specifically, plasma/serum)  Commercially prepared reagents
 Measure the concentration of a particular constituent (the  Usually provided by the suppliers of
analyte) in body fluids

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 1
 CLINICAL CHEMISTRY
SMTY1209: CLINICAL CHEMISTRY [LEC]

automated machines  Reverse Osmosis (RO) water

 In-house prepared reagents o Uses pressure to force water through a
 Requires preparation mixing of reagents semipermeable membrane
o Chemicals o Can be used for the pretreatment of
A. Analytical Grade (AR)
water and does not remove dissolved
 Suitable for most analytical procedures
 Carry designations as “AR” (analytical gases
reagent) or “ACS” (American Chemical  Ultrafiltration and Nanofiltered Water
Society) and “For Laboratory Use” or o Excellent in removing particulate matters,
“ACS Standard-Grade Reference microorganisms, and any pyrogens or endotoxins
Materials” o Through UV oxidation which removes trace
B. Ultrapure Chemicals organic materials; sterilization process at specific
• Suitable for techniques that require wavelengths; and combined with ozone treatment
extremely pure chemicals (e.g. AAS, EIA, (in the form of gas) that destroy bacteria.
MDx)  Reagent Grade Water
o Been through additional purification steps for use o Obtained by initial filter, followed by RO,
in specific procedures such as chromatography, deionization and a 0.2 mm filter
atomic absorption, immunoassays, molecular o Production of this water depends on the condition
diagnostics, etc. of the feed water
o Carry designations: “HPLC” (High performance
liquid chromatography) or “Chromatographic” CLASSIFICATION OF WATER
 Chemically Pure (CP)  Type I Water
o Impurity limitations are not stated o Used for test methods that requires minimum
o Preparation is not uniform interference such as trace metal analysis by
o Not recommended for clinical labs Frame Emission Spectrometry (FES) and Atomic
 United States Pharmacopeia (USP) & National Absorption Spectrometry (AAS)
Formulary (NF) Grade o Also used for blood gas, pH, enzyme and
o Used to manufacture drugs electrolyte analysis
o Standards are based on the criterion of not being  Type II Water
injurious to man o For analytical preparations
 Technical or Commercial Grade o Also used in reagent, QC, and standard
o Used for manufacturing preparation
o Should not be used in clinical labs  Type III Water
o Used for glassware washing but not for analysis
REFERENCE MATERIALS and reagent preparations
 Primary Standard
o Highly purified chemical that can be measured CLINICAL LABORATORY SUPPLIES
directly to produce a substance of exact known  Thermometers
concentration and purity o Analytical reactions occur at an optimal
 Secondary Standard temperature
o Substance of lower purity with concentration o Usage of circulating water or heating/cooling
determined by comparison with a primary metal blocks
standard o Integral/need to be placed
- Classifications
WATER SPECIFICATIONS  Liquid-in-glass
 Distilled water o Colored liquid or mercury encased in plastic/glass
o Purified to remove almost all organic materials material with a bulb at one end a graduated stem
through distillation where water is boiled and o Usually measures between 20°C-400°C • Types:
vaporized a. Total Immersion - The entire thermometer
o Liquid state is collected (bulb and liquid column) is exposed to the
o Tap water is not allowed temperature being measured
 Deionized water - Most commonly used in refrigerator, freezer,
o Purified by ion exchange incubator - Should be placed horizontally for the
o Water is produced using either an anion or cation temperature to be read
exchange resin and is followed by replace of the
removed ions with hydroxyl or hydrogen ions
o Removes some or all ions and is purified
previously by treated water (predistilled/distilled)
o Excellent in removing dissolved ionized solids b. Partial Immersion - Has an immersion line and
and dissolved gases should be immersed to the proper height

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 2
 CLINICAL CHEMISTRY
SMTY1209: CLINICAL CHEMISTRY [LEC]

- Used in water/cold baths • Flask is usually used in microbiology section for

- Should be used vertically preparation of agar plate
3. Graduated Cylinder
• Used to measure volumes of liquid
Immersion line – the part to be submerged for B. Pipets – Glass or plastic utensils used to transfer liquid
the temp to be read
c. Surface Thermometer - For checking
temperature on flat surfaces such as oven and
incubator
 Electronic Thermometer
o Thermistor Probe
o Fast reading, millisecond response time
 Digital Thermometer
o Used in body temperature

GLASS WARES AND PLASTIC WARES

 Commonly used in laboratory set-ups such as test tubes,
pipettes, etc.
 Glass wares: 1. Adjustable, mechanical pipettes, from left: 20-200μl, 2-20μl, 100-
A. Borosilicate 1000μl. (Has disposable tips)
o Kimax ® / Pyrex ® 2. Graded transfer pipette and electric pipette filter.
o Used for heating and sterilization 3. 25, 10, 5 and 2ml transfer pipettes.
o Most commonly used 4. Disposable tips for adjustable pipettes.
B. Aluminosilicate 5. 12 channel adjustable pipette for microplates
o Corex ® 6. (Mechanical pipette, has disposable tips) Low retention 0.5 - 10μl
o Strengthened chemically than thermally adjustable pipette.
o 6x stronger than borosilicate
7. Squeezable transfer pipettes (Disposable pipettes are used when
C. High silica
transferring biohazard samples i.e blood, etc)
o Boron
8. Digital adjustable pipette.
o High alkali resistant
D. Acid / Alkali resistant (Vycor) 9. Light guided pipetting system
o Utilized for high thermal blastic shock and
extreme chemical treatment with acids (except Pipette classification
hydrofluoric acid). Design
E. Amber Colored (Low actinic) To contain (TC) to deliver (TD)
o Protect photosensitive reagents Calibration marks/drainage characteristics
F. Lime Soda (Flint glass) Blowout Self-draining
o Made-up of soda lime glass and mixture of Ca, Si, Type
and sodium oxides. Measuring or graduated Transfer
o Poor resistance to high temperatures.
 Serologic  Volumetric
 Plasticwares
 Mohr  Ostwald-Folin
o Begins to replace glassware in the lab setting due
to high resistance to corrosion, breakage,  Bacteriologic  Pasteur pipets
disposable and flexibility.  Ball, Kolmer, Kahn  Automatic
A. Polystyrene  Micropipette macropipettes/
B. Polyethylene micropipettes
C. Polypropylene
D. Tygon Design
E. Teflon A. To Contain – does not dispose exact amount
F. Polycarbonate • Used for viscous samples
G. Polyvinyl chloride • Uses mercury as calibrating medium
• Proper use requires rinsing technique
CLASSIFICATION OF GLASS/PLASTICWARES • Hold Volume but does not dispense exact
A. Laboratory Vessels – designed to deliver (TD) or to contain (TC) volume
1. Volumetric flask B. To Deliver
 Calibrated to hold one exact volume of liquid. • Used for non-viscous samples
2. Erlenmeyer flask & Griffin Beaker • Uses distilled water as calibrating medium
• Hold different volume • Dispense the exact volume
• Used in reagent preparation

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 3
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SMTY1209: CLINICAL CHEMISTRY [LEC]

 Calibration Marks / Drainage Char. (TD)

A. Blowout Pipette
• Used in Serologic / Oswald-Folin measuring
• With etched ring / two small continuous rings

 2 rings; Note that when a substance is still present,

removing by blowing is necessary

B. Self-Draining
• Used in Volumetric / Mohr measuring
• Without marking. Drains completely

Volumetric pipet Ostwald-Folin

pipet
Size Larger Smaller
Location of the Located at the Located closer to
bulb center the delivery tip
(symmetrical)
 Type / According to Use Design To deliver To deliver
A. Measuring or Graduated Pipets
• Graduated uniformly along its length
• Designed to deliver any amount within its capacity
o Serolic Pipet
 Has graduation marks to the tip
 Blowout Pipet – continuous etched ring
o Mohr Pipet
 No graduation marks to the tip
 Self-draining – no continuous etched
ring

Mechanical and Automatic Pipets

A. Macropipet – deliver amount > 1mL
B. Micropipet – deliver amount < 1mL
C. Dilutor or Dispenser
• Automatic pipets that obtain the liquid from a
common reservoir and dispense it repeatedly
• The dispensing pipets may be bottle-top,
motorized, handheld, or attached to a dilutor.
C. Burets
B. Volumetric or Transfer Pipette • For dispensing liquid during titration E. Syringes
• Delivers an exact volume • Used to transfer small volumes in blood gas
o Ostwald-Folin Pipet analysis, chromatography, or electrophoresis
 For viscous fluids (blowout pipet)
o Volumetric Pipet
 Dessicators & Dessicants
 For aqueous solutions (self-draining)
o Uses hygroscopic (substances that take up water
when exposed to atmospheric conditions)
substance that take up water/moisture on
exposure to air

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 4
 CLINICAL CHEMISTRY
SMTY1209: CLINICAL CHEMISTRY [LEC]

 Balances
A. Top-loading Balance / Electronic Top-Loading
Balance
• For knowing the mass of substances (greater
quantity)
• Used for preparative experiments B. Filtration
• Alternative method to separate solid from liquid
• Filter materials: Paper, cellulose, polyester
fibers, and column materials.
C. Dialysis
• Method for separating macromolecules from a
solvent of smaller substancescute bullet.

SUBSUB TOPIC FORMAT

B. Analytical Balance
• For preparation of primary standards
• Measure exact mass but with lower capacities
(Operating ranges 0.01 mg to 160 g)

BASIC SEPARATION TECHNIQUE

A. 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 speed/Centrifugal force is expressed by:
 Revolution per minute (RPM)
 Relative centrifugal force (RCF) or
gravities (g)
 Centrifuged must be properly balances and free from
excessive vibrations. If only 1 tube needs to be centrifuged,
make a balancer where it has the same amount of volume
as the other tube.

EZEKIEL CRUZ. SAMANTHA CRUZ. IVAN ILAGAN. ROMINA LASCANO. KARYLLE SURIAGA. | FEU MT 2023 5