PRINCIPLES OF BASIC

TECHNIQUES
• Factors affecting Analytical process
– Concept of solute and solvent
– Units of measurement
– Chemicals
– Reference materials
– Basic techniques
– Safety
• Factors affecting Analytical process
– Concept of solute and solvent
– Units of measurement
– Chemicals
– Reference materials
– Basic techniques
– Safety
 CONCEPT OF SOLUTE AND SOLVENT
• Solution is a homogenous mixture of one or
more solutes dispersed molecularly in a
sufficient quantity dissolving solvent
– Solutes – “Analytes or measurands”
 CONCEPT OF SOLUTE AND SOLVENT
• Expressing Concentrations of Solutes
– Mole
– Molarity of a Solution
– Molality of a Solution
– Normality of a Solution
– Gram Equivalent Weight
• Factors affecting Analytical process
– Concept of solute and solvent
– Units of measurement
– Chemicals
– Reference materials
– Basic techniques
– Safety
 UNITS OF MEASUREMENT
• A meaningful measurement is expressed with
both a number and a unit.
– Number indicates how many units are contained
in the property
– Unit identifies the dimension – mass, volume, or
concentration
• International System of Units (SI)
 UNITS OF MEASUREMENT
SI Base Units

Quantity Name Symbol

Length meter m
Mass kilogram kg
Time seconds s
Electrical current ampere A
Thermodynamic temperature kelvin K
Amount of substance mole mol
Luminous intensity candela cd
Catalytic amount katal kat
• Factors affecting Analytical process
– Concept of solute and solvent
– Units of measurement
– Chemicals
– Reference materials
– Basic techniques
– Safety
 CHEMICALS
• The quality of analytical results produced by the
laboratory is a direct indication of the purity of
the chemicals used as analytical reagents
• Laboratory chemicals are available in a variety of
grades
• The solutes and solvents used in analytical work
are reagent grade chemicals, among which
water is a solvent of primary importance
 CHEMICALS
• Reagent Grade Water
• Ultrapure Reagents
 CHEMICALS
• Reagent Grade Water
• Ultrapure Reagents
 CHEMICALS
• Reagent Grade Water

CLSI Specifications for Reagent Water

CLWR
Microbiological Content 10 CFU/mL
pH NA
Resistivity ≥10 MΩcm, 25C
Silicate 0.05 mg SiO2/L
Particulate matter water passed through 0.2
μm filter
Organics water passed through
activated carbon
 CHEMICALS
• Preparation of Reagent Grade Water
– Distillation
– Ion exchange
– Reverse Osmosis
– Ultraviolet Oxidation
 CHEMICALS
• Quality, Use and Storage of Reagent grade
water
Type III Water Glassware washing
General Urinalysis
Type II Water General lab testing not
requiring type I water
(with minimum chemical or
bacterial contamination)
Type I water Trace metal analysis
Enzyme
Electrolytes
For calibration and reference
materials
 CHEMICALS
• Reagent Grade Water
• Reagent Grade or Analytical Reagent Grade
 CHEMICALS
• Chemicals that meet specifications of the ACS
are described as reagent or analytical reagent
grade
– Ultra Pure Reagents (spectograde, nanograde,
HPLC pure)
– Chemically pure
– USP and NF (drugs)
• Factors affecting Analytical process
– Concept of solute and solvent
– Units of measurement
– Chemicals
– Reference materials
– Basic techniques
– Safety
 REFERENCE MATERIALS
• Reference material is a material or substance
with one or more physical or chemical
properties that is sufficiently well established
to be used for
– Calibrating instruments
– Validating methods
– Assigning values to materials
– Evaluating compatibility of results
 REFERENCE MATERIALS
• Primary reference materials
• Secondary reference materials
• Standard reference materials (SRM)
• Certified Reference materials (CRM)
 BASIC TECHNIQUES
• Clinical laboratory supplies
– Glassware
– Plasticware
– Measuring vessels
• Methods of separating solids from liquids
– Centrifugation
– Filtration
CLINICAL LABORATORY
SUPPLIES
CHARACTERISTIC TYPES OF GLASS
 Category Type of Common or Routine uses Limitations
Material brand names
High Borosilicate Pyrex - All purpose, all types of beakers, flask - Should not be cooled
thermal with low Kimax etc too quickly after
Resistance alkaline - Can tolerate heating and sterilization heating
content for lengthy periods of time to 510C - May cloud after used
with a strong alkali
- Subject to scratching

Aluminosilic Corex - Centrifuge tubes and thermometers - Resist scratching

ate - Extremely strong and hard - Subject to some acid or
- Temperature stability to 672C; short alkali attack at
term use to 850C temperature of 100C
Vycor - Ashing and ignition techniques
- Can withstand very high temperature
(900 – 1200C) as well as drastic
changes in temperature. Most are alkali
resistant
High Silica 96% Silica - Cuvets and thermometers
- Can be used at high temperatures (900-
1200C) and withstand a sharp change
in temperature
- Can be considered optically pure
(cuvets, thermometers)

High Aluminosilic - Can be used with strong alkali and - Must be heated and
resistance ate suffer minimal attack (0.9 mg/cm2 vs cooled with care
to alkali 1.4mg/cm2 for borosilicate or 0.35 - Highest temperature for
mg/cm2 for regular aluminosilicate) safe use is 578C
CHARACTERISTIC TYPES OF
PLASTICS
Plastic Temperat Transparency Autoclavable Flexibility Usage samples
ure limit
Polystyrene 70C Clear No Rigid Disposables
(PS)
Polyethelene 80 Translucent No Excellent All-purpose
Conventional Reagent bottles
(CPE) Test tube rack
Carboys
Droppers
Linear (LPE) 120 Opaque With caution Rigid Specimen transport
containers
Reagent bottles
Polypropylene 135 Translucent Yes Rigid Screw-cap closures
(PP) Bottles
Tygon 95 Translucent Yes Excellent Tubing
Teflon FEP 205 Clear Yes Excellent Stopcocks
Translucent Wash bottles
Beakers
Polycarbonate 135 Very clear Yes Rigid All-purpose
(PC) Large reagent containers
Carboys
Test tube rack
Polyvinyl 70 Clear No Rigid Bottles/tubing
chloride (PVC)
PVC tubing 120 Yes Very
flexible
CHEMICAL RESISTANCE OF TYPES OF
PLASTIC
Plastic Chemical Resistance
Polystyren Useful with water and aqueous salt solutions. It is not
e recommended for use with acids, aldehydes and ketones,
ethers, hydrocarbons, or essential oils. Alcohols and bases can
be used, but storage beyond 24hours is discouraged
Polyethyle They have excellent chemical resistance to most substances,
ne with the exception of aldehydes, amines, ethers, hydrocarbons,
and essential oils.
For conventional polyethylene, the exceptions should also
include lubricating oil and silicones.
The usage of any of the above- named chemical groups should
be limited to 24h at room temp
Polypropyl Has the same chemical resistance as linear polyethelene
ene
Teflon This resin possesses excellent chemical resistance to almost all
chemicals used in the clinical lab
Polycarbon Very susceptible to damage by most chemicals. It is resistant to
ate water, aqueous salts, food, and inorganic acids for a long period
of time