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Introduction to
Clinical Chemistry 1
What is Laboratory Medicine?
Same as Clinical Laboratory
Refers to the discipline involved in the
selection, provision, and interpretation of
diagnostic testing that uses primarily
samples from patients
2
Scope/Uses of Laboratory Medicine
5
What is Clinical Chemistry?
Laboratory section that performs qualitative
and quantitative analyses of blood, urine,
spinal fluid, feces, calculi and other materials.
6
What is Clinical Chemistry?
Human Diseases
8
Role of Medical Technologists
9
Definition of Terms
Accuracy: ability to determine the true and
known value of as substance
Biochemical Marker: any biochemical
compound that is sufficiently altered in a
disease to serve as an aid in diagnosing or
predicting susceptibility to the disease.
Blood borne: carried or transmitted by blood
Biochemical compound= antigen, antibody,
enzyme, hormne
10
Definition of Terms
Diagnosis: the identification of the nature
of an illness or other problem by
examination of the symptoms
Pathogen: causative agent of a disease
Prognosis: an opinion, based on medical
experience, of the likely course of a
medical condition
Prognosis: forecast based on medical
experience
11
Definition of Terms
12
Summary
Treatment to correct
the abnormalities
13
Unit 2. Laboratory
Mathematics
Intended Learning Outcomes and
Content of Unit 2
Compute concentration of solutions using the different ways of expressing
concentration and using different units of measurement.
16
Quantitative Determination
Two parts: actual value and label
Actual value: a number
Label: a unit that defines the physical
quantity or dimension
Mass, length, time, volume, temperature,
concentration, activity, frequency, current
17
Unit of measurement
Systeme International dUnites (SI units):
1960
Provides a uniform system or method of
describing physical quantities (based on
metric system)
Basic units
Derived units
Selected accepted non-SI units
18
SI- adopted intenationally in 1960.
Subclassification of SI units: Basic, Derived
Derived units: derivative or a mathematical
function of one of the basic units
Selcted accepted non-SI units: non basic nor
derived units, but due to long term use, was
accepted for use
MRBPC UST 19
Taken from: Bishop et al., 7th Edition, 2013, p. 4
20
Examples:
1. 1 millimeter = ________ m
2. 2 micromoles = _______ mol
Taken from: Bishop et al., 7th Edition, 2013, p. 4 3. 5 deciliter = __________ L
4. 2 milligram = _________ kg
5. 5 microliter = _________ L 21
Added to SI units to indicate decimal fractions or
multiples of that unit.
=indicates a subunit or multiple of a basic unit
2mg=2 X10-6 kg
Larger to smaller units: move decimal to the right
Smaller to larger units: move decimal to the left
MRBPC UST 22
Basic clinical laboratory
conversions
Length, volume, weight conversions
24
Properties of Solution
Concentration
Terms to Remember:
Percent Solution Solution
Molarity Solute
Solvent
Molality Analytes
Normality
Saturation
Colligative Properties
Redox Potential
Conductivity
Ways of expressing concentration of solution: Percent solution, M, m, N
Properties of Solution
25
Percent Solution:
Weight/volume
Answer = 6.8 g
Properties of Solution
27
Percent Solution:
Weight/volume
Answer = 333.3 mL
Properties of Solution
28
Percent Solution:
Weight/volume
Properties of Solution
29
Percent solution:
volume/volume
Answer = 28.57%
Properties of Solution
30
Percent solution:
volume/volume
Properties of Solution
31
Percent solution:
volume/volume
Properties of Solution
32
Percent Solution:
Weight/weight
Properties of Solution
33
Percent Solution:
Weight/weight
Properties of Solution
34
Molar Solution or Molarity
Number of moles expressed per 1 liter of
solution
Moles = gram/molecular weight
Gram molecular weight of a substance
dissolved to a final volume of 1 liter
solution
M = moles solute
volume of solution (L)
M= grams of solute
MW x volume of solution (L)
Properties of Solution
35
Molar Solution or Molarity
Properties of Solution
36
Molar Solution or Molarity
Answer = 1.0 M
Properties of Solution
37
Molar Solution or Molarity
Properties of Solution
38
Normal solution or Normality
N = gram solute
(MW/valence) x volume of solution (L)
Valence: number of units that can combine with or replace 1 mole of hydrogen
ions for acids and hydroxyl ions for bases and the number of electrons exchange
in redox reactions Properties of Solution
39
Normal solution or Normality
MW = 98.09
Properties of Solution
40
Normal solution or Normality
MW = 110.98
Properties of Solution
41
Molarity vs Normality
Normality is always equal to or greater than
Molarity
NM
MN
Properties of Solution
42
Convert Molarity to Normality
Answer: 1 N
Properties of Solution
43
Molal Solution or Molality
m = grams of solute
MW x kg of solvent
Answer = 0.5 m
MW = 40.07
Properties of Solution
45
mg/dL to milliequivalent
In milliequivalent, equivalent weight is
expressed in milligrams
To convert mg/dL to milliequivalent per
liter (mEq/L)
mmol/L = mg/dL X 10
MW
Properties of Solution
48
Saturation
Diluted: with little solute
Concentrated: with larger quantity of solute
Saturated: with excess of undissolved solute
particles
Supersaturated: with greater concentration of
undissolved solute particles than saturated
solution
Diluted: one which has been made to a lower solute concentration per
volume of solvent
Properties of Solution
49
Colligative Properties
Vapor pressure: pressure at which the liquid
solvent is in equilibrium with the water vapor
Freezing point: temperature at which the vapor
pressures of the solid and liquid phases are the
same
Boiling point: temperature at which the vapor
pressure of the solvent reaches one atmosphere
Osmotic pressure: pressure the opposes
osmosis when a solvent flows through a
semipermeable membrane to establish
equilibrium
MRBPC UST Properties of Solution
50
Redox Potential
A measure of the ability of a solution to
accept or donate electrons
Reducing agents: substances that donate
electrons
Oxidizing agents: substances that accept
electrons
LEORA
GEROA
Properties of Solution
51
Conductivity
A measure of how well electricity passes
through a solution
Depends on the number of respective charges
of the ions present
Expressed as ohms-1 or mho
Resistivity: reciprocal of conductivity
Measure of a substances resistance to the
passage of electrical current
Expressed as ohms
Properties of Solution
52
Dilutions
represents the ratio of concentrated or stock
material to the total final volume of a solution
consists of the volume or weight of the
concentrate plus the volume of the diluent
Dilution factor: ratio of the concentrated
solution to the total solution
Take the concentration needed and divide by the
stock concentration
Dilution is always less concentrated than the original substance
because in dilution we add diluent to a more concentrated solution.
Dilution factor: always in reduced or lowest term written as fraction
or ratio 53
Dilution Factor
Make 50 mmol/L glucose solution from a 1000
mmol/L stock. What is the dilution factor?
50:1000 or 1: 20
Get 1 part of the stock and mix with 19 parts diluent
If the volume needed is specified, for example: 200
mL of 50 mmol/L glucose solution is needed.
How much of the stock will you need?
Get 10 mL of the stock and mix with 190 mL diluent
1:20::x:200
C1V1=C2V2
MRBPC UST 54
Example #1:
You have a 20 mg/dL stock cholesterol
standard. You need a 2 mg/dL standard. You
only have 0.500 mL of the stock to use and you
need 0.100 mL of the diluted standard.
Answer:
Dilution factor is 1:10, so you need 1 part stock plus 9 parts diluent.
1:10 :: x:0.1, x = 0.01 mL stock plus 0.09 mL diluent
You can also use the following:
0.1 mL of stock plus 0.9 mL diluent
0.2 mL of stock plus 1.8 mL diluent
0.3 mL of stock plus 2.7 mL diluent
0.4 mL of stock plus 3.6 mL diluent
0.5 mL of stock plus 4.5 mL diluent
Get the dilution factor first: 2:20 or 1:10 or 1 part stock plus 9 parts
55
diluent
Example #2:
MRBPC UST 57
What is the difference between
ratio and dilution?
MRBPC UST 58
Ratio and Dilution
MRBPC UST 59
Ratio and Dilution
MRBPC UST 60
Serial dilution
multiple progressive dilutions ranging from
more concentrated solutions to less
concentrated solutions
Patient sample is too small
Titer is to be established
Standard curve is to be made
Things to consider:
Total volume desired
Amount of diluent and stock available
Dilution factor
Final concentration needed
Support materials required
MRBPC UST 61
Serial dilution
Pediatric sample sent to the laboratory was only 0.5 mL
blood and upon centrifugation, only 0.2 mL serum was
obtained. Sample is to be tested for bilirubin (requiring a
total of 0.3 mL serum), for glucose (0.02 mL), albumin (0.1
mL), and total protein (0.1 mL). What will you do?
MRBPC UST 63
Specific Gravity, Density, and
Percent Purity
Density: mass per unit volume
Specific gravity: density of a substance
over density of water at a given
temperature (g/mL)
Actual concentration: specific gravity x
percent purity
MRBPC UST 64
Specific Gravity, Density, and
Percent Purity
Answer:
HCl= 35 g/mole
Actual weight = 0.52 g/mL
Molarity = 14.86 M
MRBPC UST 65
Buffers: pH and pOH
Henderson-Hasselbalch equation
pH = pKa + log (A-/HA)
pH = log (1/H+)
pH = -log (H+)
A lowercase p in front of certain letters means
negative logarithm of or inverse log of that
substance
pH + pOH = 14 If A/HA is equal to 1, pH equals pK and has the
greatest buffering capacity.
MRBPC UST 66
Scientific Notation
Uses exponential method of expressing
very large and very small numbers
Numbers are expressed as a product of
two numbers: digit term and exponential
term
Digit term: > or equal to 1 but < 10
Exponential term: written as a power of 10
MRBPC UST 67
Scientific Notation
Examples:
214 = 2.14 X 102
0.115 = 1.15 X 10-1
MRBPC UST 68
Significant Figures
Examples:
1. 1028.9 = _________
2. 213 = ___________
3. 0.000032 = _______
4. 4000 = __________
MRBPC UST 69
Significant Figures
1. All nonzero integers are significant figures.
2. Zeros used to locate decimal points are not
significant (0.0002; 0.002; 0.02 = 1 significant
figure)
3. Zeros appearing between numbers are
significant.
4. Zeros appearing at the end of a number with a
decimal are significant (18.80).
5. Zeros appearing at the end of a number without
a decimal may or may not be significant (180).
MRBPC UST 70
Significant Figures
Laboratory Application
1. Addition and Subtraction:
5.324 + 0.0031 = 5.3209 = 5.321
2. Multiplication and Division: product or
quotient can contain no more significant
digits than the least number of significant
figures in the numbers involved in the
calculation
0.8832 X 1.5 = 1.32480 = 1.3
MRBPC UST 71
Unit 3. Laboratory Safety
Intended Learning Outcomes and
Content of Unit 3
Apply laboratory safety procedures and precautions in the
performance of laboratory measurements.
Lecture:
1. Universal Precaution
2. Laboratory Hazards
2.1. Biological
2.2. Chemical
2.3. Electrical
2.4. Fire
2.5. Radiation
2.6. Others
3. Safety equipment
4. Laboratory waste management: Segregation, Storage,Treatment, and
Disposal
73
Hierarchy of Controls
Levels of dealing with laboratory hazards
1. Engineering controls
2. Administrative controls
3. Work practices
4. Personal protective equipment (PPE)
74
Mandated by the Occupational Safety and Health
Administration (OSHA)
1. Engineering controls: involve making changes to the work
environment to reduce work related hazard
=preferred over other because may make permanent
changes and do not rely on behavior
=use of chemical fume hoods and biological safety cabinets
2. Administrative: those that modify workers work
schedules and tasks in ways that minimize their exposure to
workplace hazards
=developing a chemical hygiene plan
=Developing SOP for chemical handling
MRBPC UST 75
3. Work practices: are procedures for safe and
proper work that are used to reduce the
duration, frequency or intensity of exposure to a
hazard.
=no mouth pipetting
=chemical substitution if possible (choosing a less
hazardous chemical for a specific procedure)
4. PPE: is a protective gear needed to keep
workers safe while performing their jobs
=respirators, face shields, goggles, gloves
MRBPC UST 76
Examples of Prevention Strategies
MRBPC UST 77
OSHA STANDARDS
Bloodborne pathogen standard
Formaldehyde standard
Laboratory standard
Hazard communication standard
Respiratory hazard
Air contaminants standard
Personal protective equipment standard
MRBPC UST 78
MRBPC UST 79
Standard Hazard Identification System
developed by the National Fire Protection
Association (NFPA)
=diamond shape color coded symbol
MRBPC UST 80
MRBPC UST 81
Safety
Equipment
Safety showers: deliver 30-50 gallons
of water per minute at 20-50 psi
MRBPC UST 83
Fume hoods; designed to Check air flow direction:
remove chemical fumes by placing a tissue paper
and aerosols aways from at the hood opening
the work area Do not fully open the sash
BSC: designed to provide of the hood
both clean work Chemicals stored in the
environment and hoods should not block
protection for employees airflow
working with biological Velocity at the face of the
hazards hood= 100-120 feet per
Fume hoods: check for minute
blockages regularly
MRBPC UST 84
Fume Hood
MRBPC UST 85
BSC I: minimum inward flow; no product protection
BSC II: minimum inward flow; with product protection
BSC III: maximum product protection if laminar flow is included and with two HEPA filters
MRBPC UST 86
Negative pressure flexible film
isolator
MRBPC UST 87
Types of Hazards
Chemical Hazards
Biological Hazards
Physical Hazards
Ergonomic hazards
Ionizing Radiation
Non-Ionizing Radiation
Noise
Electrical hazards
Mechanical Hazards
MRBPC UST 88
Chemical Hazards
Flammable/Combustible Chemicals:
Flammable: flashpoint below 37.8C
Combustible: flashpoint above 37.8C
Corrosive Chemicals
Reactive Chemicals
Carcinogenic Chemicals
MRBPC UST 89
Flammable and combustible: acetone, benzene,
ethanol, heptane, isopropanol, methanol, toluene,
xylene
=flammable gases=hydrogen
=flammable solids=paraffin
Corrosive: injurious to the skin or eyes by direct
contact or to the tissue of the respiratory and GI tracts
if inhaled or ingested
=acetic acid, sulfuric, nitric, HCl
=Bases:ammonium hydroxide, potassium hydroxide,
sodium hydroxide
MRBPC UST 90
Reactive chemicals=substances that under certain
conditions, can spontaneously explode or ignite or that
evolve heat or flammable or explosive gases
=some strong acids or bases react with water to
generate heat (exothermic reaction)
=mixture sof oxidizing agents such as peroxides, and
reducing agents such as hydrogen=may generate heat
and may be explosive
Carcinogenic=cancer causing agents
=benzidine
FLASHPOINT: temperature at which sufficient vapor is
given off to form an ignitable mixture with air
MRBPC UST 91
Biological Hazards
Biological Agents and
Toxins Ingestion
Bloodborne pathogens Inoculation
Research Animals Tactile
contamination
Inhalation of
infectious material
Sources:
Contact with patients, specimens of patients, supplies or materials, aerosol
materials, improperly processed blood products, inappropriate disposal of
waste products, expelling a spray from needles, centrifugation of infected
fluids, spills on laboratory counters, flaming inoculating loops
MRBPC UST 92
What to avoid in the laboratory?
Consumption of
food
Mouth pipetting
Smoking
Applying cosmetics
Drinking fluids
Leaving
unprotected any
skin, membranes, or
open cuts
MRBPC UST 93
Universal Precaution
Presumption that all human blood, tissue,
and most fluids are infectious for the
transmission of human immunodeficiency
(HIV), hepatitis B virus, and other blood
borne pathogens
Safe handling and Preventive measures
Decontamination methods
Vaccination requirement
Post exposure medication, counseling,
testing, and prophylaxis
MRBPC UST 94
Physical: Ergonomic Hazards
Cumulative trauma disorders
injuries involving the musculoskeletal and/or
nervous system in response to long term
repetitive twisting, bending, lifting, or assuming
static postures for an extended period of time
constant or excessive repetitive actions,
mechanical pressure, vibrations, or compressive
forces on the arms, hands, wrists, neck, or back
human error by pushing beyond ones limits or
when productivity limits are set too high
Repetitive pipetting, keyboard use, or resting wrists or
arms on sharp edges like laboratory counters
MRBPC UST 95
Physical: Ergonomic Hazards
Cumulative trauma
disorders
Carpal tunnel syndrome
Tendonitis
Tenosynovitis
Bursitis
Ganglion cyst
MRBPC UST 96
Carpal tunnel syndrome=compression and
entrapment of nerve from wrist to hand
Tendonitis=inflammation of the tendon
Tenosynovitis= inflammation or injury to synovial
sheath that surrounds a tendon
Bursitis=inflammation of one of the bursa of
synovial fluid
Primary contributors for repetitive strain
disorders: position/posture, applied force,
frequency of repitition
MRBPC UST 97
Physical: Ionizing Radiation
generated through
nuclear reactions, by
very high temperature,
via production of high
energy particles or due
to acceleration of
charged particles by
electromagnetic fields
Cosmic rays, x-rays,
gamma rays, beta
particles, UV
MRBPC UST 98
Physical: Non-Ionizing Radiation
Type of
electromagnetic
radiation that
does not carry
enough energy
to ionize atoms
Radiowaves,
microwaves,
infrared light,
and visible light
MRBPC UST 99
Physical: Noise
Fire Fire
Explosion Explosion
Asphyxiation Asphyxiation
Mechanical injuries Pressure build up
Embrittlement of
materials
Tissue damage
Tissue damage: thermal burns
A: ordinary
combustible
materials
B: flammable
liquids/gases
C: energized
electrical
equipment
A: paper, wood, fabric, plastic
D: reactive
D: combustible, reactive
metals like Mg, Na, K
metals MRBPC UST 103
Tetrahedron of fire
2. Indirect:
fire and explosion