REINALYN C.

LUNETA | CLINICAL CHEMISTRY 4A

HANDOUT 1 – INTRODUCTION TO CLINICAL CHEMISTRY PREFIXES USED WITH SI UNIT\?-

• CLINICAL CHEMISTRY – systematic study of SELECT

FACTOR PREFIX SYMBOL
biochemical processes associated with health and DECIMALS
disease and the measurement of the constituents in 10-18 Atto a ----
body fluids or tissues to facilitate diagnosis of 10-15 Femto f ----
disease. 10-12 Pico p ----
• Scope of Clinical Chemistry: 10-9 Nano n ----
1. Biochemistry 10-6 Micro µ 0.000001
2. Analytical chemistry 10-3 Milli m 0.001
3. Endocrinology 10-2 Centi c 0.01
4. Toxicology 10-1 Deci d 0.1
5. Immunology 100 Liter, meter, gram Basic unit 1.0
101 Deka da 10.0
6. Pharmacology
102 Hecto h 100.0
7. Computers
103 Kilo k 1,000.0
8. Instrumentation
104 Mega M ----
• Malfunction of a cell may be caused by a variety of
109 Giga G ----
factors: 1012 Tera T ----
1. Destruction by trauma or by invasive agents. 1015 Peta P ----
2. Genetic deficiency of a vital enzyme. 1018 Exa E ----
3. Insufficient supply of one or more essential
nutrients. COMPUTATION
4. Insufficient blood supply.
5. Insufficient oxygen supply. For conversion:
6. Malignancy.
7. Accumulation of waste products. • From large unit to smaller unit, move decimal to the
8. Failure of a control system. right.
9. Defect in the cellular recognition of certain • From small unit to larger unit, move decimal to the
signals. left.
• Substances measured in serum fall generally into the Ex. Convert 1.0 L (large) to µL (smaller)
following categories: Therefore 1 L (1 x 100) =? µL (micro = 10-6), So move
1. Substances normally present with a function the decimal 6 places to the right.
in the circulation. 1.0 becomes 1,000,000, so that
▪ E.g., glucose, total protein, albumin, 1.0 L = 1,000,000 µL
electrolytes, triglyceride,
cholesterol, hormones and vitamins Convert 5.5 mL (small) to dL (larger)
2. Metabolites – non-functioning waste mL (10-3) while dL (10-1), the difference between the two
products in the process of being cleared. exponent is two. Therefore, move decimal 2 places to
▪ E.g., Urea, creatinine, uric acid, the left so that, 5.5 mL = 0.055 dL
ammonia, bilirubin
LABORATORY SUPPLIES
3. Substances released from cells as a result of
cell damage and abnormal permeability or • Used for three primary purposes:
abnormal cellular proliferation. 1. Storage
▪ E.g., Enzymes such as LD, AST, 2. Measurement
ALT, CK, AMS, GGT, ALP, and ACP 3. Confinement of chemical reactions
4. Drugs and toxic substances
▪ E.g., Substances of abuse, GLASSWARES PLASTICWARES
therapeutic drugs, poisons Breakable Disposable
Reusable Highly shock-proof; shatter-proof
UNITS OF MEASUREMENT Ideal for acidic solutions Ideal for alkaline solutions
High thermal and
QUANTITY BASIC UNIT SYMBOL Relatively inert
corrosion resistant
Length Meter m Good optical properties Flexible
Mass kilogram kg Low cost For cryogenic experiments
Time Second s
Electric current ampere A GLASSWARES
Temperature Kelvin K
Luminous Intensity Candela cd • Five general types:
Amount of substance mole Mol 1. High thermal resistant glass
o Borosilicate glass with low alkali
content.
o Resists heat, corrosion, and
thermal shock.
o Most common are Pyrex, Kimax,
and Exax

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 REINALYN C. LUNETA | CLINICAL CHEMISTRY 4A

a. Corex PLASTICWARES
▪ 6x stronger than
borosilicate glass; 1. Polyolefins (polyethylenes, polypropylenes)
alumina-silicate o Unique group of resins with
glass. relatively inert chemical properties.
▪ Ideal for higher o Unaffected by acids, alkalies, salt
temperature solutions, and most aqueous
thermometers, solutions.
graduated o Polypropylene is more expensive
cylinders, and than polyethylene, therefore the
centrifuge tubes. latter is used in most disposable
b. Vycor plasticwares.
▪ Recommended o Polypropylene is sterilizable
for use in however it absorbs pigment and
application tends to become discolored.
involving high 2. Polycarbonate resin
temperature and o Twice as strong as polypropylene
drastic heat (from -100°C to 160°C)
shock and o Glass clear and shatterproof.
extreme chemical o Ideal for centrifuge tubes and
treatment with graduated cylinders.
acids and dilute 3. Tygon
alkalies. o Non-toxic clear plastic of modified
▪ Heatable to PVC (polyvinylchloride).
900°C and o Flexible and used to handle most
withstand chemicals.
downshock from o Can be steam autoclaved or
900°C to ice chemically sterilized.
water. 4. Teflon fluorocarbon resins
▪ Ideal for ashing o Almost chemically inert with high
and ignition corrosion resistance at extreme
techniques. temperatures.
2. High silica glass o For cryogenic experiments and
o Radiation resistant, good optical work at high temperatures over
qualities, and temperature extended periods (-270°C to 255°C)
capabilities. o Pure translucent white and inert.
o Ideal for high precision analytical
PIPETS
work and can also be used for
optical reflectors and mirrors. • Two general types:
3. Glass with high resistance to alkalies A. Classification according to FUNCTION
o Boron-free glassware for strong 1. Volumetric or Transfer pipet
alkali solutions and digestion with 2. Graduated or Measuring pipet
strong alkalies.
o Less thermal resistant than 1. VOLUMETRIC/TRANSFER PIPET
borosilicate glass. • Designed to deliver (TD) a fixed
o Known as “soft glass”. volume of liquid.
4. Low actinic glass • Consists of a cylindrical bulb joined
o Amber or red colored to reduce the at both ends to narrower glass
amount of light passing through the tubing.
substance. • Used for accurate measurements of
o Highly protective laboratory aliquots of non-viscous samples,
glassware for handling heat-labile filtrates, and standard solutions.
substances in the 300-500nm
range.
5. Standard flint glass
o Soda-lime glass which is composed
of a mixture of oxides of Si, Ca, and
Na.
o Poorly resistant to high temperature Calibration mark
and sudden changes in
temperature.
o Release alkali into the pipetted
liquid.

SELAUSO, ROMY JED L. BMLS 3A | 2


a. Ostwald-Folin pipet
o Similar to volumetric pipet but have
their bulb closer to the delivery tip.
o Used for measuring viscous fluids
as blood or serum.
2. GRADUATED/MEASURING PIPET
• Plain, narrow tube drawn out to a tip.
• Graduated uniformly along its length.
• Two types:
o Mohr pipet – calibrated between
two marks on the stem.
o Serological pipet – has graduation
marks down to the tip and blown
out to deliver the entire volume of
the pipet if etched ring or double
rings are evident near the
mouthpiece.
• Intended for delivery of predetermined
volumes.
• Principally used for the measurement of
reagents and are not generally
considered accurate enough for
measuring samples and standards.
a. Serological Pipet
Mouthpiece
Stem
Tip/Delivery tip
B. Classification according to DESIGN
1. To contain (TC) pipet
• Calibrated to contain a specified
amount of liquid but are not
necessarily calibrated to deliver that
exact amount.
• Also called “rinse-out” pipet.
• E.g., Sahli-Hb pipet
2. To deliver (TD) pipet
• Calibrated to deliver the amount of
fluid designated on the pipet. The
volume will flow out of the pipet by
gravity when the pipet is held in a
vertical position with its tip against
the inside wall of the receiving
vessel.
REINALYN C. LUNETA | CLINICAL CHEMISTRY 4A
C. Classification according to DRAINAGE
CHARACTERISTICS
1. Blow-out – the fluid remaining in the tip
is blown out. Usually there are two
etched rings/frosted bands near the
mouthpiece.
2. Self-draining – the fluid is allowed to
drain by gravity and the remaining fluid
in the tip is not blown out.
• Micropipets –
contain or deliver
volumes ranging
from 1-500µL.
o Also called as
lambda pipet;
lambda is
equal to 1µL.
o E.g., Lang-Levy, Kirk, Capillary,
Unopette, Semi-automatic pipet
(Eppendorf)
• Automated
dispensers – used
when adding
repeatedly a specific
volume of reagent or
diluent to a solution.
CHEMICALS
• The American Chemical Society (ACS) established
the specifications of chemicals in the analytical grade
category.
• The analytical reagent grade chemicals are of very
high purity. These are labeled Analytical Reagent
(AR) or ACS which state the actual purity of each lot.
These include chemicals such as spectrograde,
nanograde, and High-Performance Liquid
Chromatography grade.
• The analytical reagent grade chemical is used in:
1. Most analytical laboratory procedures
2. Making up reagents for spectrophotometry,
chromatography, trace metal analysis,
atomic absorption spectrophotometry,
fluorometry, and standardization procedures.
• Unites States Pharmacopeia (USP) and the
National Formulary (NF) grade chemicals are not
injurious to humans and are used to produce drugs.
They are not pure enough for use in most chemical
procedures.
• Less pure grade chemicals are referred to as purified
and technical. The chemically pure (CP) or pure
grade chemicals are not recommended for reagent
preparations.
• Technical or commercial grade reagents are
primarily used in manufacturing and should not be
used in the clinical laboratory.
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 REINALYN C. LUNETA | CLINICAL CHEMISTRY 4A

STANDARDS • Carbon dioxide free water – obtained by boiling

• Primary standard (PS) – a chemical that is the
highest purity and can be measured directly to
produce a substance of exact known concentration
and purity. The International Union of Pure and
Applied Chemistry (IUPAC) requires that PS be at
least 99.98% pure and that the working standards be
99.95% pure.
• Standard Reference Materials – are used as
primary standard materials in the clinical laboratory
purified by the National Bureau of Standards. These
are relatively expensive and should be used as a
reference for the comparison of commercially
obtained standards and reagents.
• Secondary standard – a substance of lower purity
whose concentration is determined by comparison to
a primary standard.
WATER
• Reagent grade water (RGW) – water suitable for
reagent and standard preparation. Categorized as
Type I, Type II and Type III. Generally, this can be
obtained by initially filtering it to remove particulate
matter followed by reverse osmosis, deionization, and
a 0.2mm filter or more restrictive filtration process.
USES OF REAGENT GRADE WATER
• Type I – used for procedures that require maximum
water purity such as:
a. preparation of standard solution
b. ultramicrochemical analysis
c. measurement of nanogram or subnanogram
concentration
d. tissue or cell culture
e. for trace metal, iron and enzyme analyses
• Type II – acceptable for most analytic requirements
including reagent, quality control, and standard
preparation.
• Type III – used in most qualitative measurement,
most procedures in urinalysis, parasitology and
histology, washing glasswares.
Type II water. This is used when carbon dioxide,
ammonia and oxygen may affect analysis.
• Distilled water – purified to remove almost all organic
materials using a technique of distillation in which
water is boiled and vaporized.
• Deionized water – produced from distilled water
using either an anion or cation exchange resin
followed by replacement of the removed particles with
hydroxyl or hydrogen ions respectively.
Other methods of water purification
• Reverse Osmosis – a process that uses pressure to
force water through a semi-permeable membrane
made of cellulose acetate, aromatic polyamides, or
cellulose, producing water that reflects a filtered
product of the original water. The procedure removes
90% of dissolved solids, and 98% of organic
impurities, insoluble matter and microbes. It does not
remove dissolved gases. This produces Type III
water.
• Filtration – uses semi-permeable membranes with
pore size of about 0.2µm to remove insoluble matter,
emulsified solids, pyrogens and microorganisms.
• Ultrafiltration and Nanofiltration – excellent in
removing particulate matter, microorganisms and any
pyrogens or endotoxins.
• Adsorption – uses activated charcoal, clay, silicate,
or metal oxides to remove organic impurities.
• Ultraviolet oxidation – which removes some trace
organic material or sterilization processes at specific
wavelengths, when used in combination with ozone
treatment, can destroy bacteria but may leave behind
residual products.

College of American Pathologists and National Committee

for Clinical Laboratory Standards (NCCLS) issued
minimum specifications for RGW:

Specifications Type I Type II Type III

Bacterial content
<10 <102 NA
(maximum CFU/mL)
pH NA NA NA
Resistivity (megaohms/cm
1.0 1.0 0.1
at 25°C)
Silicates 0.05 0.1 1.0
Particulate matter (filter in
0.22um NA NA
system)
Activated
carbon
Organic contaminants NA NA
filter in
system

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