Intro, Lab Math, and Safety
Intro, Lab Math, and Safety
Unit 1. Introduction to
Clinical Chemistry 1
General chemistry
Organic chemistry
HUMAN
PHYSIOLOGY
Biochemistry
Importance of
Clinical Chemistry
Carbohydrates
Lipids
Proteins
Nucleic Acids
Organ Systems:
Heart
Liver
Kidneys
Lungs
Reproductive
Nervous
Endocrine
Muscles
Skeletal
Blood
Human Diseases
MRBPINEDA UST
7/3/13
Role of Medical
Technologists
Deliver accurate and precise laboratory information
Provide reliable laboratory results
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
Definition of Terms
Summary
1ENT
Abnormalities
PAT
Disease Diagnosis
Treatment to correct
the abnormalities
MRBPINEDA UST
10
Unit 2. Laboratory
Mathematics
Maria Ruth B. Pineda, Ph.D.
Department of Medical Technology
University of Santo Tomas
Unit Conversions
Dilutions
Percent Solutions
Ratios
Normality
pH and pOH
Molarity
Significant figures
Molality
Scientific Notation
12
7/3/13
Quantitative
Determination
Qualitative Determination:
presence or absence of a certain
compound
Quantitative Determination:
exact amount of the compound
present
High or low???
13
14
Unit of measurement
SI Units
Basic units
BASE QUANTITY
NAME
SYMBOL
Length
Meter
Mass
Kilogram
kg
Time
Second
Electric current
Ampere
Thermodynamic temperature
Kelvin
Amount of substance
Mole
mol
Luminous intensity
Candela
cd
Frequency
Hertz
Hz
Force
Newton
Celsius temperature
Degree Celsius
Catalytic activity
Katal
kat
SELECTED DERIVED
Derived units
Selected accepted non-SI units
Minute (time)
(60s)
min
Hour
(3,600s)
Day
(86,400s)
(0.1 nm ! 10"10 m)
16
Liter (volume)
MRBPINEDA UST
Angstrom
15
PREFIX
SYMBOL
10-18
atto
10-15
femto
10-12
pico
10-9
nano
10-6
micro
10-3
milli
10-2
centi
10-1
deci
101
deka
da
102
hecto
103
kilo
106
mega
109
giga
1015
peta
1018
exa
Examples:
1. 1 millimeter = ________ m
2. 2 micromoles = _______ mol
3. 5 deciliter = __________ L
4. 2 milligram = _________ kg
5. 5 microliter = _________ L
17
PREFIX
10"18
atto
10"15
femto
10"12
pico
10"9
nano
10"6
micro
10"3
milli
10"2
centi
10"1
deci
101
deka
da
102
hecto
103
kilo
104
mega
109
giga
tera
Multiply by
1015
2.54peta
1018
0.39exa
SYMBOL
P
E
0.91
Meters
Yards
1.09
Gallons
Liters
3.78
Liters
Gallons
0.26
Fluid ounces
Milliliters
29.6
Milliliters
Fluid ounces
0.034
Ounces
Grams
28.4
Grams
Ounces
0.035
Pounds
Kilograms
Kilograms
Pounds
0.45
18
2.2
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Ways of expressing
concentration of solution
Temperature conversions
To convert
into
Use
Celsius (C)
Kelvin (K)
K = C +273
Celsius (C)
Fahrenheit (F)
F = (C x 1.8) + 32
Fahrenheit (F)
Celsius (C)
C = (F 32) x 0.556
As percent solution
As a molar (M) solution
As a normal (N) solution
As a molal (m) solution
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19
Percent Solution:
Weight/volume
%w/v
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20
Percent Solution:
Weight/volume
What amount of NaCl is needed to make
800 mL of 0.85% solution?
Answer = 6.8 g
22
Percent Solution:
Weight/volume
Percent Solution:
Weight/volume
23
24
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Percent solution:
volume/volume
%v/v
Percent solution:
volume/volume
Prepare 500 mL 40% alcohol from stock
absolute alcohol solution.
Answer = 200 mL stock absolute alcohol + 300
mL distilled water to make 500 mL solution
Answer = 28.57%
25
MRBPINEDA UST
Percent solution:
volume/volume
26
Percent Solution:
Weight/weight
%w/w
27
28
Percent Solution:
Weight/weight
Molar Solution or
Molarity
Number of moles expressed per 1 liter of solution
M =
grams of solute
MW x volume of solution (L)
29
MRBPINEDA UST
30
s to set up a
(Eq. 1-22)
nt solution is
sed as grams
mL of a 10%
ng approach.
0 ! 100 g
al amount)
(Eq. 1-23)
hydrochloric
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Example
1-4PART 1 BASIC PRINCIPLES AND PRACTICE OF CLINICAL CHEMISTRY
22
Normality
where V is the valence of the compound. Using this forNormality (N) is expressed as the number of equivalent
mula, Example 1-7.3 becomes
weights 1-5
per liter (Eq/L) or milliequivalents per milliliter
Example
(mEq/mL).
Equivalent
weight
is
equal
to
gmw
divided
by
0.5 M ! 2 " 1 N
(Eq. 1-34)
A solution of NaOH is contained within a Class A 1-L
the valence (V). Normality has often been used in acidvolumetric
flask filled to the calibration mark. The conbase calculations because an equivalent weight of a subExample 1-8
tent
label
reads
24togitsofcombining
NaOH. Determine
the admolarity.
stance
is also
equal
weight. Another
What is the molarity of a 2.5 N solution of HCl? This
Step
1: What
units
are ultimately
Answer: Moles
vantage
in using
equivalent
weight is needed?
that an equivalent
problem may be solved in several ways. One way is to use
weight
one substance is equal to the equivalent weight
the stepwise approach in which existing units are exper
literof(mol/L).
of
any
other
chemical.
changed for units needed. The equation is
Step 2: The units that exist are grams and 1 L. NaOH may
Molar Solution or
Molarity
Normal solution or
Normality
1 mol weight
HCl per 1 liter of
36ofggram
HCl equivalent
Number
be expressed as moles and grams. The gmw of NaOH is 2.5 Eq HCl
!
!
Example 1-7
L solution (Eq/L)
36 g HCl
1 Eq
calculated
to
equal
40
g/mol.
Rearrange
the
equation
so
Give the equivalent weight, in grams, for each substance
Equivalent
weightHCl
= molecular weight/valence
Prepare
of a 0.5and
M solution
of
that
grams
bemL
canceled
the remaining
units re" 2.5 mol/L
(Eq. 1-35)
listed
below.can400
NaCl.
flect
those
needed
in
the
answer,
which
are
mole/L.
The second approach is to use the normality-to-mo1. NaCl (gmw " 58 g, valence " 1)
=
grams
of solutenow becomes
Step 3:Answer
The equation
becomes
larity conversionN
formula.
The equation
= 11.6 g of NaCl is dissolved in
(Eq. 1-24)
of moles per
per milliliter
ance is equal
to determine
(Eq. 1-28)
(Eq. 1-29)
400
distilled water
to make 0.5 M
24 mL
g NaOH
mol
1"1)
mol
2. HCl (gmw " 36, valence
"
! 0.6
solution
L of NaCl40 g NaOH
L
(Eq. 1-26)
M ! V " 2.5 N
V"1
2.5 N
performing the approM"
" 2.5 N
(Eq. 1-36)
1
priate calculations,
the
final
answer
of
0.6
M
or
0.6
mol/L
98/2 " 49 g per equivalent weight
(Eq. 1-30)
When the valence of a substance is 1, the molarity will
is derived.
out
units
and
3.By
H2canceling
SO4 (gmw "
98,like
valence
33" 2)
34
Normal solution or
Normality
7 g H2SO4
1 Eq
1,000 mL
!
!
500 mL
49 g H2SO4
1L
" 0.285 Eq/L " 0.285 N
(Eq. 1-31)
35
Normal solution or
Normality
36
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22
Molarity vs Normality
36.5 g HCl
4.8 mol HCl
!
! 250 mL ! 1 L
mol
L
1,000 mL
" 43.8
HCl than
(Eq. 1-27)
Normality is always equal
to org greater
Molarity
Convert Molarity to
Normality
Normality
Normality (N) is expressed as the number of equivalent
weights per liter (Eq/L) or milliequivalents per milliliter
(mEq/mL). Equivalent weight is equal to gmw divided by
the valence (V). Normality has often been used in acidbase calculations because an equivalent weight of a substance is also equal to its combining weight. Another advantage in using equivalent weight is that an equivalent
weight of one substance is equal to the equivalent weight
ofofany
other
Amount
solute
perchemical.
1 kg of solvent
Molal Solution or
Molality
mol/kg (or m)
Example 1-7
0.5 M ! 2 " 1 N
Example 1-8
(Eq. 1-28)
39
mg/dL to milliequivalent
7 g H2SO4
1 Eq
1,000 mL
mg/dL X 10
! X valence
!
500 mL
49 g H2SO4
1L
MW
Other Conversions
41
(Eq. 1-31)
(Eq. 1-35)
M ! V " 2.5 N
V"1
40
M"
(Eq. 1-30)
36 g HCl
The second approach is to use the normality-to-molarity conversion formula. The equation now becomes
(Eq. 1-29)
A. What is the normality of a 500-mL solution that contains 7 g of H2SO4? The approach used to calculate molarity could be used to solve this problem as well.
Step 1: Units needed? Answer: Normality expressed as
equivalents per liter (Eq/L).
Step 2: Units you have? Answer: Milliliters and grams.
Now determine how they are related to equivalents per
In milliequivalent,
equivalent
weight
expressed in
liter. (Hint: There
are 49
g perisequivalentsee
Equation
milligrams
130 above.)
Step 3:mg/dL
Rearrange
the equation
that
like terms cancel
To convert
to milliequivalent
perso
liter
(mEq/
L) out, leaving Eq/L. This equation is
mEq/L =
1 Eq
" 2.5 =
mol/L
HCl
Answer
0.5 m
Molal Solution or
Molality
(Eq. 1-34)
(Eq. 1-33)
where V is the valence of the compound. Using this formula, Example 1-7.3 becomes
2.5 N
" 2.5 N
1
(Eq. 1-36)
Specific Gravity
mg/dL to millimoles
Density is expressed as mass per unit volume. The specific gravity is the ratio of the density of a material when
compared to the density of water at a given temperature.
The units for specific gravity are grams per milliliter.
In millimoles, molecular weight expressed in
Specific
gravity is often used with very concentrated mamilligrams
terials, such as commercial acids (e.g., sulfuric and hy To convert
mg/dL to millimoles:
drochloric
acids).
The density of a concentrated acid can also be exmg/dL
X 10
pressed mmol/L
in terms of =
an assay
or percent
purity. The actual
concentration is equal to MW
the specific gravity multiplied
by the assay or percent purity value (expressed as a decimal) stated on the label of the container.
MRBPINEDA UST
Other Conversions
42
7/3/13
Basic concentration
conversions
To convert
into
Use
%w/v
Molarity (M)
%w/v
Normality (N)
N = (%w/v x 10)/eq.wt.
mg/dL
mEq/L
Molarity (M)
Normality (N)
N = M x valence
43
44
45
46
Serial dilution
47
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48
7/3/13
Answer:
Actual weight = 0.52 g/mL
Molarity = 14.86 M
pH and pOH
Henderson-Hasselbalch equation
pH = pKa + log (A-/HA)
pH = log (1/H+)
pH = -log (H+)
pH + pOH = 14
49
50
Scientific Notation
Scientific Notation
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51
52
Significant Figures
Significant Figures
Examples:
1.1028.9 = _________
2.213 = ___________
3.0.000032 = _______
4.4000 = __________
MRBPINEDA UST
Examples:
214 = 2.14 X 102
0.115 = 1.15 X 10-1
53
7/3/13
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
END OF UNIT 2
Quiz next meeting!
Bring calculator and periodic table of elements.
56
Lecture:
Laboratory:
1. Universal Precaution
1. Hand washing
2. Laboratory Hazards
2. Cleaning of glassware
2.1. Biological
3. Disinfection of working areas
2.2. Chemical
4. Laboratory waste management
2.3. Electrical
2.4. Fire
2.5. Radiation
2.6. Others
3. Safety equipment
4. Laboratory waste management:
Segregation, Storage,Treatment, and
Disposal
57
58
Examples of Prevention
Strategies
Hierarchy of Controls
1. Engineering controls
2. Administrative controls
3. Work practices
59
MRBPINEDA UST
TABLE 1-16
TABLE 1-17
PART 1
TABLE 1-15
10
7/3/13
OSHA STANDARDS
Bloodborne pathogen standard
Formaldehyde standard
Laboratory standard
Hazard communication standard
Respiratory hazard
Air contaminants standard
01Arneson (F)-01
12/18/06
6:26 PM
Page 4
MRBPINEDA UST
62
Chapter 1
Safety Regulations
In the clinical chemistry laboratory, local, state, and federal regulations, including
the Occupational Safety and Health Act (OSHA), provide guidelines for safe operation of testing processes. Regulations include guidelines for operating safety
equipment and identifying, handling, and storing chemical hazards. Table 11 outlines the types of safety equipment that may be necessary to operate a clinical
chemistry laboratory safely. Table 12 outlines proper identification of chemicals
within the laboratory through the use of material safety data sheet (MSDS) sys-
TABLE 1-1
Safety Equipment
Safety
Equipment
Equipment
Glasses or goggles
Work shields
Gloves
Coat or apron
FUME HOOD
Fume Hoods
Explosion-proof
refrigerators
Compressed gas
storage
Storage cabinets
FIRE EXTINGUISHERS
Class A
Class B
Class C
Class ABC
Pressurized water
Carbon dioxide
Dry chemical
Dry chemical
SAFETY SHOWER
EYEWASHES
SPILL KITS
Fume Hood
Biosafety Cabinets
MRBPINEDA UST
Use
Unbreakable eye shields that sur- Protects exposed skin and clothes
that may be worn outside the
round the eye area
laboratory
Spatter protection for exposed
skin
Latex or vinyl cover for hands
Cover for clothes that will be
worn outside the laboratory
STORAGE UNITS
64
Description
fume hoods
have improved
containment
however, it is safer
to work with the
sash lowered
65
Avoid Clutter, Do
not cover airfoil
MRBPINEDA UST
Put equipme
to rear of ho
66
11
7/3/13
Mechanism of BSC
MRBPINEDA UST
67
Types of BSC
78
AIRFLOW PATTERN
75
No
2, 3
No
Class II Type A
75
No
2, 3
Yes
Type B1
100
Yes
(low levels/
volatility)
2, 3
Yes
Type B2
100
Yes
2, 3
Yes
Type B3
100
Yes
2, 3
Yes
Class III
NA
Yes
3, 4
TYPE
APPLICATIONS
RADIONUCLIDES/
TOXIC CHEMICALS
MRBPINEDA UST
BIOSAFETY
LEVEL(S)
PRODUCT
PROTECTION
70
Source: Centers for Disease Control and Prevention and the National Institutes of Health. Biosafety in microbiological and biomedical laboratories.
4th ed. Washington, D.C.: U.S. Government Printing Office, 1999: Table 1, Comparison of Biological Safety Cabinets.
IFPM, Linear feet per minute
*Glove panels may be added and will increase face velocity to 150 Ifpm; gloves may be added with an inlet air pressure release that will allow
work with chemicals/radionuclides.
BIOLOGIC SAFETY
Nitrile gloves, for example, offer a wider range of compatibility with organic solvents than do latex gloves. Lab
coats, preferably with knit-cuffed sleeves, should be full
length and buttoned and made of liquid-resistant material. When performing manipulations prone to splash
hazards, the lab coat should be supplemented with an
impermeable apron and/or sleeve garters, constructed of
suitable material to guard against the substances. Proper
footwear is required; shoes constructed of porous materials, open-toed shoes, or sandals are considered ineffective against spilled hazardous liquids.
Respirators may be required for various procedures in
the clinical laboratory. Whether used for biologic or
chemical hazards, the correct type of respirator must be
used for the specific hazard. Respirators with highefficiency particulate air (HEPA) filters must be worn
when engineering controls are not feasible, such as when
working directly with patients with tuberculosis (TB) or
when performing procedures that may aerosolize specimens of patients with a suspected or confirmed case of
TB. Training, maintenance, and written protocol for use
of respirators are required according to the respiratory
protection standard.
Each employer must provide (at no charge) lab coats,
gloves, or other protective equipment to all employees
who may be exposed to biologic or chemical hazards. It
is the employers responsibility to clean and maintain all
PPE. All contaminated PPE must be removed and properly disposed of before leaving the laboratory.
General Considerations
All blood samples and other body fluids should be collected, transported, handled, and processed using strict
precautions. Gloves, gowns, and face protection must be
used if splashing or splattering is likely to occur.
Consistent and thorough hand washing is an essential
component of infection control.
Centrifugation of biologic specimens produces finely
dispersed aerosols that are a high-risk source of infection. Ideally, specimens should remain capped during
centrifugation. As an additional precaution, the use of a
centrifuge with an internal shield is recommended.
Start Up
1. Turn off
ultraviolet light
as soon as you
enter the room.
2. Turn on all
blowers and
cabinet
illumination
lights.
3. Open sash to
appropriate
working level.
3. Allow five
minutes of
operation to
purge system;
check flow alarm
system audio and
visual alarm
function if so
equipped.
4. Decontaminate
readily accessible
interior surfaces
with a
disinfectant
appropriate for
the agents or
suspected agents
present.
Shut down
1. Decontaminate
and remove all
items from
interior work
area.
MRBPINEDA UST
2. Decontaminate
readily accessible
interior surfaces
with a
disinfectant
appropriate for
the agents or
suspected agents
present.
3. Allow five
minutes of
operation to
purge system.
71
4. Turn off
cabinet
illumination
lights and blower.
Close the sash.
5. Turn on
ultraviolet light.
If light is not
used, leave
blower on and
sash open
Spills
used in field
work where BSC
is not feasible/
appropriate to
install
MRBPINEDA UST
Any blood, body fluid, or other potentially infectious material spill must be cleaned up, and the area or equipment
must be disinfected immediately. Cleanup includes the
following recommendations:
72
12
7/3/13
Types of Hazards
Chemical Hazards
Chemical Hazards
Flammable/Combustible Chemicals:
Biological Hazards
Physical Hazards
Ergonomic hazards
Ionizing Radiation
Non-Ionizing Radiation
Noise
Corrosive Chemicals
Reactive Chemicals
Carcinogenic Chemicals
Electrical hazards
TABLE 1-15
Mechanical Hazards
74
Biological Hazards
Biological Agents and
Toxins
Ingestion
Consumption of food
Inoculation
Mouth pipetting
Bloodborne pathogens
Tactile contamination
Research Animals
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
MRBPINEDA UST
75
Smoking
Applying cosmetics
Drinking fluids
Personal protective equipment (PPE; barriers that physically
Leaving
unprotected
any a hazard)
separate
the user from
skin, membranes, or
open cuts
MRBPINEDA UST
76
Ha
Cle
Av
Re
Im
Job
Or
No
Wa
Pu
Saf
Bio
Sp
Vo
Ce
Bio
Me
Co
Sen
No
Go
Ma
Fac
Pro
Eye
Ch
TABLE 1-16
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
MRBPINEDA UST
77
78
lo
f
m
p
n
p
t
7/3/13
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
MRBPINEDA UST
79
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
MRBPINEDA UST
81
MRBPINEDA UST
80
Physical: Non-Ionizing
Radiation
Type of
electromagnetic
radiation that does
not carry enough
energy to ionize
atoms
Radiowaves,
microwaves,
infrared light, and
visible light
MRBPINEDA UST
82
Mechanical Hazards
Physical: Noise
Anything that has the
potential to cause hearing
loss
Centrifuge
Autoclave
Exposure to an equivalent
sound pressure level of
more than 85 dB over an
8-hour period workday
MRBPINEDA UST
Physical: Ergonomic
Hazards
Homogenizers
Glasswares
83
MRBPINEDA UST
84
14
Compressed
Gases Hazards
CLASS OF FIRE
Cryogenic
Material Hazards
Fire Hazards
Fire
Fire
Explosion
Explosion
Asphyxiation
Asphyxiation
Mechanical injuries
Pressure build up
B: flammable liquids/
gases
C: energized electrical
equipment
Embrittlement of
materials
MRBPINEDA UST
85
86
Tetrahedron of fire
Uninhibited
reaction
OPERATION
ABC
Pressurized Water
Dry Chemical
PULL
PIN
A IM
NOZZLE
ABC
SQUEEZE
BC
Dry Chemical
TRIGGER
Carbon Dioxide
SWEEP
Class C Fires
Halon
NOZZLE
ABC
BC
Metal X
Halon
Dry Chemical
FIGURE 3-4. Proper use of fire extinguishers. (Adapted with permission from the Clinical and Laboratory Safety
Department, The University of Texas Health Science Center at Houston.)
MRBPINEDA UST
Electrical
Hazards
7/3/13
Carbon Dioxide
Tissue damage
TYPE OF EXTINGUISHER
Class D Fires
Class B Fires
D: reactive metals
Class A Fires
A: ordinary
combustible materials
ciated with the use of electrical appliances and equipment. Hazards of electrical energy can be direct and result
in death, shock, or burns. Indirect hazards can result in
fire or explosion. Therefore, there are many precautionary
procedures to follow when operating or working around
electrical equipment:
Electrical injuries:
1. Direct:
death, shock, or
burns
Fuel
Oxygen
otection
2. Indirect:
fire and explosion
Heat
FIGURE 3-3. Fire tetrahedron.
MRBPINEDA UST
g Radiation
88
Waste Management
Classification of Fires
Fires have been divided into four classes based on the nature of the combustible material and requirements for
extinguishment:
try of Fire
Recycling/Resource recovery
End of Unit 3
um
MRBPINEDA UST
APPROXIMATE
WAVELENGTH
PROTECTIVE MEASURES
1 cm!
Radiofrequency
coil in
ICPmass
MRBPINEDA
UST
spectrometer
Engineered
shielding and
89
posted pacemaker warning
3 m3 mm
Engineered shielding
750 nm0.3 cm
Containment and
appropriate warning labels
400750 nm
4400 nm
cabinets
warning labels
MRBPINEDA,
RMT, MSMT
UST
90
MRBPINEDA UST
15