LESSON 1

UNITS OF MEASURE ▪ Ultrapure

• Quantitative Laboratory  Have been put additional purification
o Actual test value steps for use in specific procedures
o Units such as chromatography, atomic
• Systeme International d’ Unites (SI) absorption, immunoassays, molecular
o 3 Classes diagnostics, standardization, or other
▪ Base techniques that require extremely pure
chemicals.
▪ Derived
▪ United States Pharmacopeia (USP);
▪ Supplemental
National Formulary (NF)
 Used to manufacture drugs
 Chemicals in this group may be pure
enough for use in most chemical
procedures
▪ Chemically Pure / Pure Grade
 The impurity limitations are not
stated, and that preparation of these
chemicals is not uniform.
 Not recommended that clinical
laboratories use these chemicals for
reagent preparation unless further
purification or a reagent blank is
included
▪ Technical or Commercial
 are used primarily in manufacturing
and should never be used in the
clinical laboratory

REFERENCE MATERIALS
• Standards
Figure 1-1 Systeme International Unites
o Primary
▪ highly purified chemical that can be
measured directly to produce a
REMEMBER substance of exact known concentration
• ANALYTES – moles of solute / volume of and purity. (ACS) - American Chemical
solution (mmol/L) Society
• ENZYMES – International Unit per liter (U/L) o Secondary
• pH SCALE – measurement of Hydrogen ion ▪ substance of lower purity with
concentration. concentration determined by comparison
• LITER – for ref volume with a primary standard.

National Institute of Standards and Technology

REAGENTS • Developed standard reference materials (SRMs)
• Chemicals for use in clinical chemistry laboratories.
o Grades of Purity • They are assigned a value after careful analysis.
▪ Analytic Reagent But they may not possess the purity equivalent of
 Suitable for use in most analytic a primary standard.
laboratory procedures • Used when producing calibrator and standard
materials.

PASCUAL INTRODUCTION TO CLINICAL CHEMISTRY | 1

 WATER SPECIFICATIONS
• The most frequently used reagent in the
laboratory.
• Distilled water - solely purified by distillation
• Deionized water - water purified by ion
exchange
• RO water - pumps water (pressure) across a
semipermeable membrane
• Ultrafiltration / Nanofiltration
o Excellent in removing particulate matter,
microorganisms and any pyrogens or
endotoxins
• UV oxidation - removes some trace organic
material.
• Reagent Grade Water – laboratory requirement
o Initially filtered it to remove particulate matter
followed by reverse osmosis deionization,
and a 0.2-mm filter or more restrictive
filtration process
o Quality determination
• Measurements of resistance (devoid ions, poor
conductor of electricity), pH, colony counts (for
assessing bacterial contamination) on selective
and nonselective media for the detection of
coliforms, chlorine, ammonia, nitrate or nitrite,
iron, hardness, phosphate, sodium, silica,
carbon dioxide, chemical oxygen demand
(COD), and metal detection.
• SIX CATEGORIES
o Clinical Laboratory Reagent Water (CLRW)
o Special Reagent Water (SRW)
o Instrument Feed Water
o Water supplied by Method Manufacturer
o Autoclave and Wash Water
o Commercially bottled purified water
• WATER PARAMETERS
o Microbiological counts
o pH sensitivity
o silicate
o Particulate matter
o Organics
• CATERGORIES OF WATER PURITY
o Type 1 – used to test methods requiring
minimum interference, such as trace metal,
iron, and enzyme analyses
PASCUAL
o Type 2 - acceptable for most analytic
requirements, including reagent, quality
control, and standard preparation,
o Type 3/autoclave wash water - acceptable
for glassware washing but not for analysis or
reagent preparation.
SOLUTION PROPERTIES
• Solute
o substance that is dissolved in a liquid
o biologic solutes are also known as analytes
• Solvent
o liquid in which the solute is dissolved
CONCENTRATION
• Percent solution is expressed as the amount of
solute per 100 total units of solution.
o weight per weight (w/w)
o volume per volume (v/v)
o weight per volume (w/v)
Molarity (M)
• is expressed as the number of moles per 1 L
of solution.
Molality (m)
• represents the amount of solute per 1 kg of
solvent.
Normality (N)
• defined as the number of gram equivalent
weights per 1 L of solution
COLLIGATIVE PROPERTIES
• Those properties related to the number of solute
particles per solvent molecules
OSMOTIC PRESSURE
• The pressure that opposes osmosis when a
solvent flows through a semipermeable
membrane to establish equilibrium between
compartments of differing concentration.
VAPOR PRESSURE
• The pressure exerted by the vapor when the
liquid solvent is in equilibrium with the vapor.
FREEZING POINT
• The temperature at which the first crystal (solid)
of solvent forms in equilibrium with the solution.
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BOILING POINT
• The temperature at which the vapor pressure of
the solvent reaches atmospheric pressure
(usually one atmosphere).
TEMPERATURE CONVERSIONS
• Centigrade to Kelvin= C + 273
• Centigrade to Fahrenheit= (C X 1.8) + 32
• Fahrenheit to Centigrade= (F -32) X 0.556
GLASSWARES
PIPET CLASSIFICATION
• Design
o TC - do not deliver that same volume when
the liquid is transferred into a container
o TD – labware will deliver that amount.
• Drainage characteristics
o Blowout - a continuous etched ring or two
small, close, continuous rings located near
the top of the pipet
o Self-draining - user allows the contents of
the pipet to drain by gravity
Figure 1-2 Pipet Classification
• Type
o Measuring or graduated - capable of
dispensing several different volumes.
▪ Serologic - has graduation marks to the
tip and is generally a blowout pipet
▪ Mohr – does not have graduation to the
tip (SD), the tip should not be allowed to
touch the vessel while the pipet is
draining
▪ Micropipette - with a total holding
volume of less than 1 ML, may be
designed as either a Mohr or serologic.
o Transfer - designed to dispense one volume
without further subdivisions, bulblike
enlargement in the pipet stem easily
distinguishes the Ostwald-Folin and
volumetric
PASCUAL
▪ Volumetric – dispense or transfer
aqueous solutions and is always self-
draining, has the greatest degree
accuracy and precision and should be
used when diluting standards,
calibrators, or quality-control material.
▪ Ostwald-folin - used with biologic fluids
having a viscosity greater than that of
water. (BO)
▪ Pasteur pipets – do not have calibration
marks and are used to transfer solutions
or biologic fluids without consideration of
a specific volume.
▪ Automatic pipet - most routinely used
pipet in today’s clinical chemistry
laboratory.
 Fixed volume - associated with only
one volume
 Variable - able to select different
volumes however, only one volume
may be used at a time.
▪ Fully automated/ self-operating,
semiautomatic, completely manually
operated.
General Types
• Air displacement - relies on a piston for suction
creation to draw the sample into a disposable tip
that must be changed after each use.
• Positive displacement - operates by moving
the piston in the pipet tip or barrel, much like a
hypodermic syringe, does not require a different
tip for each use.
• Dispenser – automatic pipets that obtain the
liquid from a common reservoir and dispense it
repeatedly.
CATERGORIES OF GLASSWARE
• Kimax/Pyrex (borosilicate)
o Most commonly used.
o used for heating and sterilization purposes
o has a high degree of thermal resistance, low
alkali content and free from Magnesium lime-
zinc group of elements, heavy metals,
arsenic, and antimony
o Strain point: 515oC (Pyrex)
• Boron-free glassware/ Soft glass
o Has high resistance to alkali.
o Its thermal resistance is LESS compared to
borosilicate glass
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• Corex (Aluminosilicate)
o a special alumina-silicate glass that has
strengthened chemically than thermally.
o 6x stronger than borosilicate.
• Vycor (Corning)
o Utilized for high thermal, drastic heat shock
and extreme chemical treatment with acids
and dilute alkali.
o Can be heated to 900oC.
• Flint glass
o Made up of soda-lime glass and a mixture of
Calcium, silicon, and sodium oxides.
o Has poor resistance to high temperature
LABORATORY VESSELS
• Flasks
o Volumetric - calibrated to hold one exact
volume of liquid (TC), has a round, lower
portion with a flat bottom and a long, thin
neck with an etched calibration line.
o Erlenmeyer – designed to hold different
volumes rather than one exact amount, are
often used in reagent preparation, has a wide
bottom that gradually evolves into a smaller,
short neck.
• Beakers
o Griffin beaker – has a flat bottom, straight
sides, and an opening as wide as the flat
base, with a small spout in the lip.
• Graduated cylinders
o Long, cylindrical tubes usually held upright
by an octagonal or circular base.
o Has calibration marks along its length and is
used to measure volumes of liquids.
• Burets
o looks like a wide, long, graduated pipet with
a stopcock at one end, used to dispense
used to dispense
• Dessicants
o (Drying agents) to keep other chemicals from
becoming hydrated.
• Dessicators
o a closed, airtight chamber.
• Balances
o Electronic balances - single-pan balances
that use an electromagnetic force to
counterbalance the weighed sample’s mass.
PASCUAL
o Analytic balances – required for the
preparation of any primary standards, also
known as a substitution balance, has a
single pan enclosed by sliding transparent
doors.
• Centrifuge
o Generally used to separate serum or plasma
from the blood cells as the blood samples are
being processed
o Centrifugal force variables; mass, speed
(rpm), radius
▪ Expressed in terms of relative centrifugal
force (RCF) or gravities (g)
 RCF = 1.118 x 10-5 x r x (rpm)2
where;
• 1.118 x 10-5 is a constant
• r is the radius in centimeters
Figure 1-3 Laboratory Vessels
NOTES TO REMEMBER
• In draining of fluid in receiving vessel, pipets
should be in VERTICAL position with the TIPS
AGAINST THE SIDE OF THE RECEPTACLE.
• For volumetric TD pipet, it should NOT BE
SHAKEN OR HIT AGAINST THE WALL of the
container during draining.
• Distilled water- calibrating medium for TD pipets
• Mercury- calibrating medium for TC pipets.
• Gravimetric and Spectrometric methods- used to
verify pipet volume accuracy and precision.
• 0.1% phenol red soln’ in distilled water- used to
compare the reproducibility of brands of pipet tips.
• Class A pipets do not require recalibration.
• Plastic pipette - made with polypropylene.
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