ANALYTICAL CHEMISTRY • Analysis of steel during its production permits
adjustment in theconcentrations of such
LECTURE AND LABORATORY TOPIC
1. INTRODUCTION TO ANALYTICAL
CHEMISTRY
2. QUANTITATIVEANALYTICAL METHODS
3. SELECTING ANDHANDLING REGEANTS
AND OTHER CHEMICALS
4. APPARATUS OFANALYTICAL
CHEMISTRY
Lesson 1: INTRODUCTION
Analytical chemistry is ameasurement
scienceconsisting of a set ofpowerful ideas
andmethods that are useful inall fields of
science, engineering, and medicine.
ROLES OF ANALYTICAL CHEMISTRY
Qualitative analysis establishes the chemical
identity of the species in the sample.
Quantitative analysis determines the relative
amounts of these species, oranalytes, in
numerical terms.
APPLICATION OF ANALYTICAL
CHEMISTRY:
• Quantities of hydrocarbons, nitrogen oxides,
and carbonmonoxide present in automobile
exhaust gases are measured todetermine the
effectiveness of emission-control devices.
• Quantitative measurements of ionized calcium
in blood serumhelp diagnose parathyroid
disease in humans.
• Quantitative determination of nitrogen in foods
establishes theirprotein content and thus their
nutritional value.
• One of the first questions that must be
considered in theselection process is the level of
elements as carbon, nickel, and chromiumto
achieve a desired strength, hardness, corrosion
resistance, and ductility.
QUANTITATIVE ANALYTICAL METHODS
We compute the results of a typical quantitative
analysis from two measurements. One is the
mass or the volume of sample being analyzed.
The second measurement is of some
quantitythat is proportional to the amount of
analyte in the sample suchas mass, volume,
intensity of light, or electrical charge.
ANALYTICAL METHODS
GRAVIMETRIC METHODS - Mass of the
analyte or some compound chemically related to
it.
VOLUMETRIC METHODS - volume of a
solution containing sufficient reagent to react
completely with theanalyte.
ELECTROANALYTICALMETHODS - measure
electrical properties such as potential, current,
resistance, and quantity of electrical charge.
SPECTROSCOPICMETHODS -
electromagnetic radiation and analyte atoms or
molecules or the emission ofradiation by
analytes.
SEQUENCE OF QUANTITATIVE ANALYSIS
1. CHOOSING A METHOD
• The essential first step in any quantitative
analysis is theselection of a method.
accuracy required.
• A second consideration related to economic
factors is thenumber of samples that will be
analyzed.
• Finally, the complexity of the sample and the
number ofcomponents in the sample always
influence the choice ofmethod to some degree.
2. ACQUIRING THE SAMPLE
The second step in a quantitative analysis is to
acquire thesample. To produce meaningful
information, an analysis mustbe performed on a
sample that has the same composition asthe
bulk of material from which it was taken. When
the bulk is large and heterogeneous, great effort
is required to get arepresentative sample.
✓ HETEROGENEOUS - A material is
heterogeneous ifits constituent parts can
bedistinguished visually or with theaid of
a microscope. Coal, animal tissue, and
soil are heterogeneous.
✓ ASSAY- An assay is the process
ofdetermining how much of agiven
sample is the materialby its indicated
name.
✓ SAMPLING - Sampling is the process
ofcollecting a small mass of amaterial
whose compositionaccurately
represents thebulk of the material
beingsampled.
3. PROCESSING OF SAMPLE
The third step in an analysis is to process the
sample. Under certain circumstances, no
sample processing is required priorto the
measurement step. The first step in processing
thesample is often the preparation of a
laboratory sample.
a. PREPARINGLABORATORY SAMPLE
A solid laboratory sample is ground todecrease
particle size, mixed to ensurehomogeneity, and
stored for various lengths of time before analysis
begins.
Liquid samples present a slightly differentbut
related set of problems during thepreparation
step. If such samples are allowedto stand in
open containers, the solvent mayevaporate and
change the concentration ofthe analyte.
b. DEFINING REPLICATE SAMPLES
Most chemical analyses are performed
onreplicate samples whose masses or
volumeshave been determined by
carefulmeasurements with an analytical balance
orwith a precise volumetric device
c. PREPARING SOLUTIONS: PHYSICAL &
CHEMICALCHANGES
Most analyses are performed on solutions ofthe
sample made with a suitable solvent.Ideally; the
solvent should dissolve the entiresample,
including the analyte, rapidly andcompletely.
The conditions of dissolutionshould be
sufficiently mild that loss of the analyte cannot
occur.
4. ELIMINATING INTERFERENCES
Species other than the analyte that affect the
finalmeasurement are called interferences, or
interferents. Ascheme must be devised to isolate
the analytes frominterferences before the final
measurement is made.
SEQUENCE OF QUANTITATIVE ANALYSIS
(to be discussed)
5. CALIBRATING & MEASURING
CONCENTRATION
6. CALCULATING RESULTS
7. EVALUATING RESULTS
*Notezzzzz*
✓ Use of different measurements and tools
✓ ANALYTICAL CHEM - NASA spending space
exploration to MARS - in order to study mars
(sending out rovers to collect samples like rocks,
soil etc.) to know what the composition of planet
itself
✓ ANALYTICAL CHEM - central science
✓ Qualitative - composition
✓ Quantitative - "quantity" / exact amount of
species in numerical terms
✓ Analytes - Components of the sample that
are determined
✓ EX: [Sample - soil Analyte - what you would
be determining Sample - Blood Analyte - level of
oxygen, carbon dioxide, glucose]
✓ Quantitative Analysis - there 2 measurements
1. Mass of the volume of the sample.
2. Quantity that is proportional to the amount of
analyte in the sample / unit depends on the use
of method
✓ Methods (Units): Mass, Volume, Intensity of
light, Electrical charge
✓ Final Answer = Numerical + Unit
✓ Analytical Methods
✓ Gravimetric Method - Mass is the Unit (g, kg)
Disadvantage would be time consuming just to
produce results and very tedious (lab could take
6hrs+)
✓ Volumetric Method - Volume of a solution that
would react completely with the analyte
✓ Titration - Is also a process of neutralization
(salt + water) how to achieve end color like
adding reagent (basic, acidic) to reach then
computation to see the final solution to measure
analyte
✓ Indicators - would tell you when to stop
✓ Electro analytical Method - applying electricity
to measure potential, current, resistance and
quantity charge
✓ Spectroscopic Method - emission of radiation
by analytes
✓ Miscellaneous Groups of Method - use of
radioactive decay, rate of refraction etc. (mostly
done in work with additional training)
✓ Sequence of Quantitative Analysis
1. Choosing a method - sometimes relatively
difficult, you would be needing experience
(intuition) to choose a method (usually senior
analysts know a lot more about what to do)
q1. Know the level of accuracy (closeness to the reability (triplication) what numbers you get, you Lesson 2: CALCULATIONS USED IN
true value) have to get the average for final answer ANALYTICAL CHEMISTRY

q2. Economic factors - operational cause & 3. Preparing solutions: physical & chemical IMPORTANT UNIT OF MEASUREMENT
steps / depends on number of samples (& changes - made with suitable solvent, solvent
machines) should dissolve the entire sample (rapidly & SI UNITS:
completely)
q3. Complexity of sample and number of Physical Name of Abbreviation
components to be test to know if you need to + 4. Eliminating Interferences Quantity Unit
Mass Kilogram kg
or - procedures or when to improvise
✓ eliminate substances that can interfere (aka Length Meter m
2. Acquiring the sample Time Second s
interferences/interferents - species that causes
Temperature Kelvin K
an error that can affect the final measure in an Amount of Mole mol
✓ performed in some sample (same analysis by enhancing or attenuating the Substance
composition as the bulk of material which it was quantity being measured Electric Ampere A
taken) should not be biased. Current
✓ there should be a scheme that isolates the Luminous Candela cd
✓ Heterogeneous - can be distinguished analytes from interferences Intensity
visually (sandy, clay, rocky etc)
5. Calibrating & Measuring concentration
✓ Assay - process of determining how much PREFIXES FOR UNITS
given sample by its indicated name ✓ Process determining the proportionality
between analyte concentration & measured Prefix Abbreviation Multiplier
✓ Sampling - it’s the most difficult, process of Yotta- Y 1024
quantity
Zetta- Z 1021
collecting small mass of a material whose
Exa- E 1018
composition accurately represents the bulk of ✓ Specific - 1 and only analyte
Peta- P 1015
the material (ex. biological sources) Tera- T 1012
✓ Selective - few analytes
Giga- G 109
3. Processing sample
Mega- M 106
6. Calculating Results
Kilo- K 103
✓ know the measurement of a sample
Hecto- h 102
✓ easy because computers do the work base
Deca- da 101
1. Preparing laboratory sample - solid laboratory on raw experimental data but there are cases or Deci- d 10-1
sample (mixed to ensure homogeneity, stored some problems, that’s why we learn manual Centri- c 10-2
for various lengths of the before analysis begins, computation/procedure Milli- m 10-3
disadvantage would be absorption or Micro- μ 10-6
desorption) or liquid (disadvantage would be 7. Evaluating Results Nano- n 10-9
evaporation, gases would dissolve into liquid) Pico- p 10-12
✓ Delta check - checking previous results aka Femto- f 10-15
2. Defining replicate sample - portion of material evaluate Atto- a 10-18
that approximately same size + same time + Zepto- z 10-21
same way have a copy, with replication it would Yocto- y 10-24
improve quality of the results and measure
THE DISTINCTION BETWEEN MASS AND
WEIGHT
• Mass - It is an in variant measure of the
quantity of matter in an object.
• Weight -It is the force of attraction
between an object and its surroundings,
principally the earth
THE MOLE
1. The mole (abbreviated mol) is the SI unit for
the amount of a chemical substance. It is always
associated with specific microscopic entities
such as atoms, molecules, ions, electrons, other
particles, or specified groups of such particles as
represented by a chemical formula.
2. It is the amount of the specified substance
that contains the same number of particles as
the number of carbon atoms in exactly 12 grams
of 12C. This important number is Avogadro’s
number NA = 6.022 x 1023
3. The molar mass of a substance is the mass in
grams of 1 mole of that substance. For example,
the molar mass of formaldehyde CH2O is
CALCULATING THE AMOUNT OF A
SUBSTANCE IN MOLES
1. Find the number of moles of benzoic acid
(M=122.1 g/mol) that are contained in 2.00 g of
the pure acid.
Solution:
2. Find the number of moles of 25.0 g of Na2
SO4(142.0 g/mol)
Solution:
SOLUTIONS AND THEIR CONCENTRATIONS
SOLUTION
• A homogenous mixture of two or more
substances with each substance
retaining its own chemical identity
• A solution contains two or more
components: a solvent and one or more
solutes.
• Solutions used in laboratories and
clinical settings are most often liquids,
and the solvent is nearly always water.
Solvent – is the component of a solution that is
present in the greatest amount
Solute – is the component of a solution that is
present in a lesser amount relative to that of the
solvent
CONCENTRATION OF SOLUTIONS
• refers to the weight or volume of the
solute present in a specified amount of
solvent or a solution
• THREE BASIC TYPES OF
SOLUTIONS:
A. Percent solutions
B. Molar solutions
C. Normal solutions
A. PERCENT SOLUTIONS
• Amount of solute in a solution can be
measured as a percentage of the total
volume of the solution
• Expressed as equal parts per hundred
or the amount of solute per 100 total
units of solution
• Three expressions of percent solution:
1. Percent by mass (mass-mass percent or
%w/w)
- is the mass of solute in a solution divided by
the total mass of solution, multiplied by 100 (to
put the value in terms of percentage).
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑥 100
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
2. Percent by volume (volume-volume Practice Question:(volume-volume percent CALCULATING THE AMOUNT
percent or %v/v) or %v/v)
OF SOLUTE OR SOLVENT
- is the volume of solute in a solution divided by What is the percent-by-volume concentration if a
the total volume of solution, multiplied by 100. 2mL of concentrated HCl is diluted with 80mL IN A GIVEN PERCENT
distilled water?
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 SOLUTION
𝑥100
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 Given:
Practice Question:
3. Mass-volume percent (%w/v) 2mL conc. HCl
Normal saline solution (NSS) is used to dissolve
- is the mass of solute in a solution (in grams) 80mL dH2O drugs for IV use which is 0.9% w/v NaCl in
divided by the total volume of solution (in water. How many grams of NaCl is needed to
milliliters), multiplied by 100. Solution: prepare a 50mL NSS?

𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 Given: Volume of solution = 50mL

𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒(𝑔) Percent by volume = 𝑥 100
𝑥 100 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 (𝑚𝐿)
Mass-volume percent = 0.9% NaCl solution
2 𝑚𝐿 𝑐𝑜𝑛𝑐. 𝐻𝐶𝑙 (NSS)
Practice Question: (mass-mass percent or 𝑥 100
2 𝑚𝐿 𝑐𝑜𝑛𝑐. 𝐻𝐶𝑙+80 𝑚𝐿 𝑑𝐻2𝑂
%w/w)
Solution: Mass-volume percent
2 𝑚𝐿 𝑐𝑜𝑛𝑐. 𝐻𝐶𝑙
What is the percent-by-mass concentration of 𝑥 100
82 𝑚𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
sucrose in a solution made by dissolving 7.5g of = 𝑥 100
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
sucrose in 86.5g of water? = 2.44% HCl solution
𝑥
Given: Practice Question: Mass-volume percent = 0.9 𝑔 𝑥 100
35 𝑚𝐿
(%w/v)
7.5g sucrose 0.9%9(35 𝑚𝐿)=𝑥100
What is the concentration of a 200 mL solution = 0.9%(35 𝑚𝐿) 𝑥(100)
=
86.5g water containing 1.8g of NaCl? 100 100

Solution: Given: 1.8g NaCl X = 0.32g NaCl

𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 200mL solution B. MOLAR SOLUTIONS

Percent by mass =
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
𝑥 100
Solution: Mass-volume percent • solution containing one gram molecular
7.5𝑔 𝑠𝑢𝑐𝑟𝑜𝑠𝑒 weight (one mole of the solute in one
𝑥 100
7.5𝑔 𝑠𝑢𝑐𝑟𝑜𝑠𝑒+86.5𝑔 𝑤𝑎𝑡𝑒𝑟 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 (𝑔) liter solution) of the substance per liter
= 𝑥 100
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 (𝑚𝐿) of the solution
7.5𝑔 𝑠𝑢𝑐𝑟𝑜𝑠𝑒
𝑥 100 𝑚𝑜𝑙 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
94𝑔 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 1.8 𝑔 𝑁𝑎𝐶𝑙 • 𝑀= 𝑥 100
= 𝑥100 𝐿 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
200 𝑚𝐿 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
= 7.98% sucrose solution
= 0.9% NaCl solution or 0.9g/mL NaCl
solution
 Practice Question: 20𝑔 𝑁𝑎𝑂𝐻
40𝑔
H2SO4-2 NH4OH-1 FeC3-
𝑥 1.50 𝐿
𝑚𝑜𝑙 Fe(+3) x
Determine the molarity of a solution containing Cl(-1) =3
4.35 moles of KMnO4 dissolved in enough water Molecular weight: H3PO4-3 Ba(OH)2-2
to give 750 mL solution
NaOH= 23 + 16 + 1 Practice Question:
Given:
= 40g/mol Determine the normality of a solution
4.35 moles KMnO4 containing 15g KCl dissolved in enough
= 0.33M NaOH solution water to give 0.20L solution. (K-39.10;
750mL solution
Cl-35.45)
C. NORMAL SOLUTIONS
𝑚𝑜𝑙 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 Given:
Solution: 𝑀 = 𝑥 100
𝐿 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 • least likely to be encountered of the four 15g KCl
concentration expressions to be 0.20L solution
4.35 𝑚𝑜𝑙𝑒𝑠 𝐾𝑀𝑛04
𝑀= 𝑥 100 encountered in the clinical laboratories, Solution:
750 𝑚𝐿
but is often used in chemical titrations 𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
𝑁= 𝑥 100
= 5.8M KMnO4 and chemical reagent classification 𝐸𝑊 𝑜𝑓 𝐿 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
• The number of gram equivalent weight 15𝑔 𝐾𝐶𝑙
1𝐿 =75.44 𝑥 0.2𝑜 𝐿
Conversion:750 𝑚𝐿 𝑥 = 0.75 𝑚𝐿 per 1 L of solution.
1000 𝑚𝐿
𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒 = 1.01N KCl solution
• 𝑁 = 𝐸𝑊 𝑜𝑓 𝐿 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥 100
• Equivalent Weight = MW x valence Valence:
𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
• 𝑀 = 𝑀𝑊 𝑜𝑓 𝐿 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥 100
KCl = K(+1) x Cl(-1) = 1
IDENTIFYING THE VALENCE OF ACIDS, Equivalent weight:
BASES, AND SALTS
Practice Question:
KCl = (39.10 + 35.45) x 1
• ACIDS – count the number of Hydrogen
Determine the molarity of a solution containing
ions
20g NaOH dissolved in enough water to give = 74.55(1)
• BASES – count the number of
1.50L solution. (Na-23; O-16, H-1)
Hydroxide ions = 74.55
• SALTS – multiply the absolute value of Relationship of Normality and Molarity
Given:
the ions
• Normality is ALWAYS equal or greater than
20g NaOH ACIDS BASES SALTS
molarity of
HCl-1 NaOH-1 NaCl-
1.50L Na(+1) x that
Cl(-1)=2 compound
𝑔 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
Solution 𝑀 = 𝑀𝑊 𝑜𝑓 𝐿 𝑖𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥 100 HNO3 KOH-1 CaCl2-
Ca(+2) x
Cl(-1) =2
Practice Questions:
a) What is the molarity of a 2N NaCl solution?
b) What is the normality of a 5M H2SO4
solution?
Solutions:
a) M = Normality/valence
= 2/1
= 2M NaCl solution
b) N = Molarity x valence
=5x1
= 10N H2SO4 solution
DILUTIONS
• Represents the ratio of concentrated or
stock material to the total final volume of a
solution and consists of the volume or
weight of the concentrate plus the volume of
the diluent, with the concentration units
remaining the same.
• In the molar, normal or percentage
solutions, the amount of solute contained in
a given volume of solution is equal to the
product of volume times the concentration.
• Whenever the solution is diluted, the
volume is increased and its concentration is
decreased but the total amount of solute
remains unchanged.
• C1V1 = C2V2
Practice Question: by the supplier, and the results
are printed on the container
What is the initial volume of a 40% label.
formaldehyde diluted to prepare 100mL of
10%formaldehyde solution? SPECIAL-PURPOSE
REAGENT CHEMICALS
Given: Chemicals that have been
prepared for a specific
V1 =? C1 = 40%
application are also available.
V2 = 100mL C2 = 10%
Solution:C1V1 = C2V2 RULES FOR HANDLING REAGENTS AND
SOLUTIONS
(40%)𝑉1=(10%)(100𝑚𝐿)
= 40%𝑉1 (10%)(100𝑚𝐿) We observe the following rules to prevent
=
40% 40%
the accidental contamination of reagents
1000 and solutions:
V1 =
40
1. Select the best grade of chemical
V1 = 25mL available for analytical work. Whenever
possible, pick the smallest bottle that is
Lesson 3: SELECTING
sufficient to do the job.
ANDHANDLINGREGEANTS ANDOTHER
CHEMICALS 2. Replace the top of every container
immediately after removing reagent. Do not
CLASSIFYING CHEMICALS
rely on
REGEANT GRADE
3. Hold the stoppers of reagent bottles
Reagent-grade chemicals between your fingers. Never set a stopper
conform to the minimum on a desktop.
standards set forth by the
Reagent Chemical Committee of 4. Unless specifically directed otherwise,
the American Chemical Society never return any excess reagent to a
(ACS) and are used whenever bottle. The money saved by returning
possible in analytical work.
excesses is seldom worth the risk of
contaminating the entire bottle.
PRIMARY-STANDARD GRADE
5. Unless directed otherwise, never insert
Primary-standard reagents spatulas, spoons, or knives into a bottle
have been carefully analyzed that contains a solid chemical. Instead,
shake the capped bottle vigorously or tap it
gently against a wooden table to break up
an encrustation. Then pour out the desired
quantity. These measures are occasionally
ineffective, and in such cases a clean
porcelain spoon should be used.
6. Keep the reagent shelf and the laboratory
balance clean and neat. Clean up any
spills immediately.
7. Follow local regulations concerning the
disposal of surplus reagents and solutions.
APPARATUS OF ANALYTICAL
CHEMISTRY
1. CLEANING AND MARKING
OFLABORATORY WARE
results can be sufficiently vigorous to cause
partial loss of the solution.
• Some unwanted substances can be
eliminated during evaporation. Organic
constituents can frequently be eliminated
from a solution by adding sulfuric acid and
heating to the appearance of sulphur
trioxide fumes (in a hood). This process is
known as wet ashing.
MEASURING MASS
In most analyses, an analytical balance
must be used to measure masses with high
accuracy. Less accurate laboratory
balances are also used for mass
measurements when the demands for
reliability are not critical.
ELECTRONIC ANALYTICAL BALANCE
• The pan rides above a hollow metal
cylinder that is surrounded By a coil that fits
over the inner pole of a cylindrical
permanent Magnet.
• An electric current in the coil produces a
magnetic field that supports or levitates the
cylinder, the pan and indicator arm, and
whatever load is on the pan.
• The current is adjusted so that the level of
the indicator arm is in the null position when
the pan is empty. Placing an object on the
pan causes the pan and indicator arm to
move downward, thus increasing the
amount of light striking the photocell of the
null detector.
PRECAUTIONS IN USING ANALYTICAL

• A chemical analysis is usually performed
in duplicate or triplicate. Each vessel that
holds a sample must be marked so that its
contents can be positively identified.
• Every beaker, flask, or crucible that will
contain the sample b must be thoroughly
cleaned before being used. The apparatus
should be washed with a hot detergent
solution and then rinsed—initially with large
amounts of tap water and finally with
several small portions of de ionized water.
2. EVAPORATING LIQUIDS
• Evaporation is frequently difficult to control
because of the tendency of some solutions
to overheat locally. The bumping that
3. MEASURING MASS
• An analytical balance has a maximum
capacity that ranges from 1 g to several
kilograms and a precision at maximum
capacity of at least 1 part in 105.
• A macro balance is the most common
type of analytical balance, and it has a
maximum load of 160 to 200 g and a
precision of 0.1 mg.
• A semi micro analytical balance has a
BALANCE
maximum load of 10 to 30 g and a precision
of 0.01 mg.
• A micro analytical balance has a
An analytical balance is a delicate
maximum load of 1 to 3 gm and a precision
instrument that you must handle with care.
of 0.001 mg, or 1 μg. Observe the following general rules for
working with an analytical balance
regardless of make or model:
1. Center the load on the pan as well as
possible.
2. Protect the balance from corrosion.
3. Observe special precautions for the
weighing of liquids.
4. Consult your instructor if the balance
appears to need adjustment.
5. Keep the balance and its case
scrupulously clean.
6. Always allow an object that has been
heated to return to room temperature before
weighing it.
7. Use tongs, finger pads, or a glassine
paper strip to handle dried objects to
prevent transferring moisture to them.
Lesson 4: Good Laboratory Practice
GOOD LABORATORY PRACTICE
✓ A process to certify that every step of the
analysis is valid it compasses the application
of Total Quality Management (TQM) in the
clinical laboratory set up which requires the
following:
1. Quality assurance
2. Quality control
3. Provision of an SOP
4. Validation of data / results generated in
the laboratory
5. Participation in proficiency testing
procedures
PARAMETERS INVOLVED IN TQM
1. Quality Assurance – a system of ensuring
the integrity of data in the laboratory.
2. Quality control – tools utilized to sustain the
integrity of data in the laboratory.
3. Validation – process that ensures that the
performance characteristic of a particular
procedure in the laboratory meets the standard
set by regulatory guidelines.
4. SOP – Standard Operating Procedures (a
guide containing the totality of the various
examination performed in the laboratory).
5. Proficiency Testing – A series of
examination required and prescribed a particular
national reference laboratory for the legal
purpose of validation.
PHASES OF VALIDATION PROCESS
A. Pre-analytical
B. Analytical
C. Post analytical
A. Pre Analytical Phase - Activities covering
all procedures done prior to the actual
testing of the target analyte
E.g. procurement of the sample, labelling and
processing of the sample
B. Analytical Phase - Main focus of validation
should be done properly and correctly
Covers all activities done during the actual
testing procedure
E.g. pipetting, reaction of the sample with the
reagents, incubation, measurement of the target
analyte using specific parameters like the
absorbance, generation / computation of results
C. Post Analytical Phase - Covers all
activities rendered after the testing
procedure
E.g. transcription of results, evaluation and
distribution of test results
TYPES OF VALIDATION PROGRAMS
1. External or Inter-laboratory Validation
Program – Proficiency testing given /
rendered by a national reference
laboratory such as the following:
a. National Kidney & Transplant Institute –
National reference laboratory hematology
b. RITM – National reference laboratory for
Microbiology, virology and tropical medicine
c. East Avenue Medical Center – National
reference laboratory for chemistry drug of
abuse and toxicology
d. Philippine Heart Center –National
reference laboratory for histopathology and
cardiovascular markers / monitoring
2. Internal or Intra-laboratory Validation
Program - process of establishing reference
values in the laboratory.
PRACTICAL CONSIDERATIONS IN A
VALIDATION PROGRAM
1. Specimen collection and handling
2. Reagents and equipment
3. Methods and procedures
4. Standards and controls
5. Qualified personnel
TOOLS OF VALIDATION/QUALITY CONTROL
1. The use of reference materials like the
following:
a. Standard solution - composed of known
constituent / component and of known
concentration used to compare for accuracy
✓ TYPES OF STANDARD SOLN: Primary
and Secondary Standard
b. Control solution – composed of known
constituent / component but of an unknown
amount / concentration used to compare for
precision
✓ TYPES OF CONTROL SOLN: Pooled Sera
/ Commercially Prepared
c. Blank solution – solution intended to set
the reading of the machine / calibrating the
machine. It is used to compare for accuracy
✓ TYPES OF BLANK SOLN: Water Blank and
Reagent Blank
2. The application of statistical analysis to
identify patterns of errors
VARIATIONS
✓ Errors in Quality Control and / or validation
✓ The fundamental basis of any statistical
analysis
SOME IMPORTANT TERMS
✓ The mean, also called the arithmetic mean
or the average, is obtained by dividing the
sum of replicate measurements by the
number of measurements in the set.
✓ The median is the middle value in a set of
data that has been arranged in numerical
order.
✓ Precision describes the reproducibility of
measurements—in other words, the
closeness of results that have been obtained
in exactly the same way.
✓ Accuracy indicates the closeness of the
measurement to the true or accepted value
and is expressed by the error.
TYPES OF ERRORS OBSERVED DURING
VALIDATION
1. Random / Indeterminate Error - Due to
unpredictable cause / origin is not possibly
determined ( affects the precision of results)
TYPES OF RANDOM ERRORS Ex. Co-precipitation of impurities, impurities of
reagent, unstable specimen, side reactions, slow
a. Inherent Random Error (Inherent to the or incomplete reactions
equipment)
CORRECTIONS OF METHODIC ERRORS:
Ex: fluctuation in temperature, slight variation in
function of instrument ✓ Proper method development / procedural
changes
b. Limited Random Error (limitations of
observation) c. Instrumental Error - (Due to the
instrument itself)
Ex: fatigue of the eye of the observer
Ex. Variation in temperature, Contamination of
CORRECTION OF RANDOM ERRORS:
equipment, Power fluctuations, Component
✓ Use of high grade equipment failure, Damaged parts of the instrument
✓ Careful use of the equipment
CORRECTIONS OF METHODIC ERRORS:
2. Determinate/ Residual/ Systematic Error - ✓ Calibration
Magnitude can be determined; tangible ✓ Proper instrument maintenance
(affects the accuracy of results)
3. Gross Error / Blunders - Lead to the
TYPES OF DETERMINATOR ERRORS
formation of outliers / requires statistical
a. Personal Error ( Due to carelessness, techniques to be rejected
prejudice and color acuity problems )
Ex. Spilling of small portions of sample during
Ex. Use of dirty apparatus, improper calibration, the transfer of liquids to the container, “overrun
Poor sample preparation (Continuation –sample endpoint”, instrument breakdown, loss of crucial
personal error), Misreading of data, Personal sample
Bias, Improper calculations, Incorrect listing of
weights and Over titration
CORRECTION OF PERSONAL ERRORS:
✓ Proper training and experience on the part
of the analyst / clinical laboratory officer /
Medical Technologist
b. Methodic Error (Due to the method which
cannot be eliminated unless a change is
made with it)
 Laboratory: analytical Vycor Pipette Classifications
instruments
Corning CALIBRATIONS MARKS/DESIGN

- Utilized for high thermal, 1.To deliver (TD)- delivers exact amount it
GENERAL AND COMMON LABORATORY drastic heat shock and holds into a container
EQUIPMENT extreme chemical 2. To contain (TC)- holds the particular volume
treatment with acids and but does not dispense the exact volume
Types of Glassware DRAINAGE CHARACTERISTICS
dilute alkali
1. Borosilicate glass (Pyrex and kimax) Flint glass
1. Blowout
2. Boron-free glassware (soft glass) - with continuous etched ring on top of the
- Made up of soda-lime
3. Corex (corning) pipet
glass and a mixture of
4. Vycor (corning) - exact volume is obtained when the last
Calcium, silicon, and
5. Flint glass drop is blown out
sodium oxides
- Poor resistance to high 2. Self-draining
temp - without etched ring on top of the pipet
Borosilicate Glass
Types of Plastic Wares - liquid is allowed to drain by gravity
Pyrex and Kimax TYPES of pipette
• Polyolefins
- Most commonly used - POLYETHYLENE TRANSFER PIPETTE
for heating and - PROLYPROPYLENE
1. Volumetric Pipette (self-draining)
sterilization purposes
- For non-viscous fluid
- Characterized by a high • Fluorocarbons
Steps:
degree of thermal - TEFLON
resistance and has low - PDVF 1. Using mechanical suction
alkali content. 2. Wipe off outside of pipette with gauze
Boron-free glassware • Engineering Resins 3. Adjusting the meniscus
- NYLON 4. Drain into receiving vessel
Soft glass
- ACETAL
- High resistance to alkali. - POLYCARBONATE 2. Ostwald Folin (for viscous fluid)
- Its thermal resistance is less as compared to - POLYSTYRENE - With etched ring
borosilicate glass - POLYPHENYLENE OXIDES 3. Pasteur pipette
- Commonly made from plastic
Corex • Labware Plastics - Transfers fluid without
- CORIAN consideration of a specific volume
Corning - EPOXY RESINS 4. Automatic micro or macro pipettes
- ABS - A pipet associated with only one
- Special alumina-silicate glass
- POLYETHIRAMIDE volume (fixed volume) or able to
select different volumes
GRADUATED OR MEASURING PIPETTE
1. Serological pipette
- With graduations to the tip
- Blowout pipette
2. Mohr pipette
- Without graduations to the tip
- Self-draining pipette
3. Micropipettes
1) Sahli-Hellige pipet
2) Lang-Levy pipet
3) RBC and WBC pipets
4) Kirk and Overflow pipets
Steps:
1. Keeping your fingers on the end of the
pipette, gently move it to the waste
container.
2. Touch the tip to the inside of the container,
lift your finger off the end and allow the
liquid to drain out of the pipette.
3. Hold the pipette in this position for a few
seconds after it stops draining.
CLASSIFICATION ACCORDING TO MECHANISM
AIR-DISPLACEMENT
- It relies on piston for suction
creation to draw the sample into a
disposable tip
- Piston does not come in contact
with the liquid
POSITIVE DISPLACEMENT
- It operates by moving the piston in
the pipet tip or barrel, like
hypodermic syringe
- It does not require a different tip
for each use
DISPENSER/DILUTOR
- It obtains liquid from a common
reservoir and dispense it
repeatedly
- It combines sampling and
dispensing functions
Other Measuring Glassware
- Beaker
- Erlenmeyer flasks
- Volumetric flasks
- Graduated cylinder
Classification of solutions
◉ Based on Miscibility
1. Miscible - if two liquids dissolve in each
other in any proportion as in water and
alcohol
2. Partially Miscible – when two liquid
components form a single phase when
mixed in certain proportions but form two
Introduction to Analytic
phases when mixed in different proportions
Techniques
like benzene and water
Solutions 3. Immiscible – when two components are
insoluble in each other like water and
• A solution is a mixture of 2 or more mercury
substances in a single phase. ◉ Based on Saturation
• One constituent is usually regarded as the
1. Saturated solution - contains the maximum
SOLVENT and the others as SOLUTES.
quantity of solute that dissolves at that
Parts of a solution
temperature.
◉ SOLUTE – the part of a solution that is being 2. Unsaturated solution - contains less than
dissolved (usually the lesser amount) the maximum amount of solute that can
dissolve at a particular temperature
◉ SOLVENT – the part of a solution that dissolves
3. Supersaturated solutions - contain more
the solute (usually the greater amount)
solute that a solvent can dissolve at a given
◉ Solute + Solvent = Solution temperature
Supersaturated solutions are unstable. The
supersaturation is only temporary, and usually
accomplished in one of two ways:
1. Warm the solvent so that it will dissolve more,
then cool the solution
2. Evaporate some of the solvent carefully so that
the solute does not solidify and come out of
solution.
◉ Based on Heat Absorbed or evolved in formation
1. Exothermic solution – when there is
evolution of heat during its formation, thus
when the components of this solution are
mixed, the solution becomes hot, like NaOH
in water
2. Endothermic solution – when there is
absorption of heat during its formation,
thus when the components of this solution
are mixed, the solution becomes cold
◉ Based on Relative Amount of Solute Present
1. Dilute solution – a solution which contains a
relatively small amount of solute
2. Concentrated solution – a solution which
contains a relatively large amount of solute
◉ Based on the concentration in relation to a
certain standard
1. Isotonic solution – solution with the same
concentration as the standard Normal
Saline Solution (0.85 – 0.9% NaCl is isotonic
with the concentration of salt in the blood)
2. Hypotonic Solution – solution whose
concentration is lower than the standard
3. Hypertonic Solution – solution whose
concentration is greater than the standard
Properties of solutions
1. Particles in a solution are non-settling.
2. Particles in a solution pass through filters
3. Solutions are clear even when colored.
4. Particles in a solution are diffusible
Diffusion - Movement of a fluid from an area of
higher concentration to an area of lower
concentration. The particles will mix until they
are evenly distributed.
5. Solutions can undergo osmosis.
Osmosis- as the passage of water molecules
from a region of their higher concentration to a
region of their lower concentration, through a
partially permeable membrane.
◉ Colligative Properties
- Properties determined by the number of particles
in solution rather than the type of particles.
• Vapor Pressure
• Boiling Point
• Freezing Point
• Osmotic Pressure
◉ Boiling Point - of a liquid is the water temperature
at which the vapor pressure of the liquid equals the
environmental pressure surrounding the liquid
- This shows how water molecules are able to
break the forces of attraction i.e. the
hydrogen bonds to each other and escape
as the gas molecule. This is what is
happening inside the gas bubble as it is
rising to the surface to break and release
the water gas molecules.
6. Boiling point of solutions of a non-volatile
compound is always higher than in pure
solvent.
◉ Boiling-point elevation
- describes the phenomenon that the boiling
point of a liquid (a solvent) will be higher
when another compound is added, meaning
that a solution has a higher boiling point
than a pure solvent. This happens whenever
a non-volatile solute, such as a salt, is added
to a pure solvent, such as water.
7. Vapor pressure of a solution is always less
than the vapor pressure of a pure solvent
◉ The Freezing point of a pure (unmixed) liquid
- is essentially the same as the melting point
of the same substance in its solid form and
may be regarded as the temperature at
which the solid and liquid states of the
substance are in equilibrium.
◉ Freezing-point depression
- describes the phenomenon that the
freezing point of a liquid (a solvent) is
depressed when another compound is
added, meaning that a solution has a lower
freezing point than a pure solvent.
- This happens whenever a solute is added to
a pure solvent, such as water.
- The phenomenon may be observed in sea
water, which due to its salt content remains
liquid at temperatures below 0°C, the
freezing point of pure water.
Osmotic Pressure
- is the hydrostatic pressure produced by a
difference in concentration between
solutions on the two sides of a surface such
as a semipermeable membrane.

8. Osmotic pressure is higher in solutions of
higher concentrations.
Solubility
- A property of a substance which allows it to
form uniform mixtures with other
substance or the weight of a substance
dissolved by a given weight of volume of
solvent at a given temperature.
1. Soluble or very soluble – when a given
solute is readily soluble in a given amount
of solvent e.g. sugar in water
2. Moderately soluble – when a given solute in
a given amount of solvent with the aid of
outside factor such as stirring or shaking
e.g. sodium sulfate in water
3. Slightly soluble – when the solute is partially
dissolved in a given amount of solvent eg.
CaO in water
4. Insoluble – when a given solute does not
dissolve in a given amount of water e.g.
sand in water
General Rules for Solubility of Compounds
◉ Gases
- Heating in general decrease the solubility of
a gas in liquids
- Increased pressure results in increased
solubility of a gas in a liquid
◉ Liquids
- Relative solubilities of two liquids in each
other are determined by its degree of
similarity
- Liquids of low molecular polarity are readily Solubilities of Solids vs Temperature
miscible with each other
- Heat increases the miscibility of liquids • Solubilities of several ionic solid as a
◉ Solids function of temperature. MOST salts have
greater solubility in hot water.
- Solids vary in their solubility in liquids • A few salts have negative heat of solution,
- All Na, K, NH4 compounds are soluble (exothermic process) and they become less
- All compounds containing halides except of soluble with increasing temperature.
Ag, Hg and Pb are soluble
- All compounds containing nitrates, acetates
and chlorates are soluble Temperature & the Solubility of Gases
- All sulfate except Ba, Pb and Sr are soluble
- The solubility of gases DECREASES at higher
while Ag and Ca are slightly soluble
temperatures
- Most oxides, hydroxides, phosphates,
carbonates, sulfides except Na, K are
insoluble Henry’s Law
- The effect of partial pressure on solubility of
Factors Affecting Solubility gases
- At pressure of few atmosphere or less,
1) Nature of Solute / Solvent. - Like dissolves
solubility of gas solute follows Henry Law
like
which states that the amount of solute gas
2) Temperature –
dissolved in solution is directly proportional
i) Solids/Liquids- Solubility increases
to the amount of pressure above the
with Temperature Increase K.E.
solution.
increases motion and collision
between solute / solvent.
ii) gas - Solubility decreases with
c=kP
Temperature Increase K.E. result in
gas escaping to atmosphere.
c = solubility of the gas (M)
3) Pressure Factor -
i) Solids/Liquids - Very little effect Solids and
Liquids are already close together, extra
k = Henry’s Law Constant
pressure will not increase solubility.
ii) gas - Solubility increases with Pressure.
P = partial pressure of gas
Increase pressure squeezes gas solute into Henry’s Law Constants (25°C), k
solvent.
N2 8.42 •10-7 M/mmHg
O2 1.66 •10-6 M/mmHg
CO2 4.48•10-5 M/mmHg
Henry’s Law & Soft Drinks
- Soft drinks contain “carbonated water” –
water with dissolved carbon dioxide gas.
- The drinks are bottled with a CO2 pressure
greater than 1 atm.
- When the bottle is opened, the pressure of
CO2 decreases and the solubility of CO2
also decreases, according to Henry’s Law.
- Therefore, bubbles of CO2 escape from
solution.
Factors Which Speeds Up the Rate of
Solubility
• Size of Solute
- Pulverization or grinding of a solid
to fine powder will increase area to
the solvent
• Agitation
- Shaking or stirring bring out about
circulation of the solvent and
maximum contact between solute
and solvent
• Temperature
- Heating increases the solubility of
most solids
• Nature of the reactants
- Among the halogens, Fluorine is
the most active and combines with
Hydrogen with explosive violence
• Close Contact
- This favors reaction. If there is
greater contact and collision
among the molecules, there is
increased reaction velocity.
• Temperature
- Chemical reactions proceed more
rapidly in higher temperature.
• Catalysts
- This speeds up the rate of the
reaction. Some catalysts form
intermediated product with one of
the reactants while others are only
contact catalysts. Negative
catalysts retard the rate of the
reaction.
• Concentration
- The more molecules in a definite
volume, the greater will the speed
of reaction.
- is the measure of how much of a
given substance there is mixed
with another substance.
- This can apply to any sort of
chemical mixture, but most
frequently the concept is limited to
homogeneous solutions, where it
refers to the amount of solute in a
substance.
Measuring Concentrations
Qualitative Description
1. Dilute or Weak
- solutions of relatively low
concentration
2. Concentrated or Strong
- solutions of relatively high
concentration
Quantitative Description
• Molarity(M): moles solute / Liter solution
• Molality* (m) - moles solute / Kg solvent
• Normality (N) – equivalent weight solute
Liter solution
• Mole Fraction(A) - moles solute / total
moles solution
Mole
• SI unit for the amount of chemical
substance
• Is the number of atoms in exactly 0.012kg
of 12 Carbon (approximately 6.022x10^23)
Mole= weight in grams of compound
---------------------------------------------
Gram molecular weight
Molarity / formal concentration
• number of moles of a substance per liter of
solution
Molarity = moles of solute
--------------------------
Liters of solution
Molality
• number of moles of a solute per kg. of
solvent (not total solution)
• the masses of solute and solvent do not
change with temperature as long as neither
one is allowed to evaporate
Molality = moles of solute
-----------------------
Kg of solvent
Normality
N = grams of solute
-------------------
EW x volume (L)

EW = GMW
-----------------
Valence
Percent composition and density
• Percentage of solute in a total volume of
solution
• Percentage of component in a mixture or
solution express as weight percent
Weight % = mass of solute
------------------------------------- x100
mass of total solution
Volumetric analysis
• General term for quantitative chemical
analysis in which the amount of substance
is determined by the measurement of the
volume that the substance occupies.
• Referred to as “titration”
Titration
• Equivalence point
- Is a theoretical point reached when
the amount of added titrant is
chemically equivalent to the
amount of analyte in the sample.
• End point
- The point in titration when the
physical change occurs that is
associated with the condition of
chemical equivalence
• Titrant 1. Laboratory hazards
- solute that has a known
concentration and identity
• Analyte
- solution that has a known volume
and identity
• Indicator:
- Is a compound with a physical
property (usually color) that
changes abruptly when titration is
complete.
- The change is caused by the
disappearance of the analyte or
appearance of excess titrant.
4 types of titration
• Acid-base titration- use acid and base as
reactants
• Complexometric titration- involves metals
• Oxidation-reduction- reducing and
oxidizing agent
• Precipitation- formation of precipitate
Back titration
• Direct titration is sometimes not feasible
due to:
1. Reaction rate is slow
2. No suitable indicator
3. The color change is slow of delayed
4. The end point is far from the equivalent point
• A base or reagent is added to a known
quantity- greater than the amount required
for acid neutralization
Bloodborne Pathogens
- This standard applies to all exposure to
blood or other potentially infectious
materials in any occupational setting.
Hazard Communication
- It defines hazardous substances and
provides guidance for evaluating and
communicating identified hazards.
Laboratory standards
- Requires the appointment of a chemical
hygiene officer and the development of a
chemical hygiene plan to reduce or
eliminate occupational exposure to
hazardous chemicals.
Universal Precaution
- an approach to infection control in which all
human blood, tissue, and most fluids are
handled as if known to be infectious for the
human immunodeficiency virus (HIV),
hepatitis B virus (HBV), and other
bloodborne pathogens.
2. Laboratory Hazard and Standards
Biological hazards
- Biological hazards expose an unprotected
individual to bacteria, viruses, parasites,
other biological entities that can result in
injury. o
- Exposure occurs from ingestion,
inoculation, tactile contamination, or
inhalation of infectious material from
patients or their body fluids/tissues,
supplies or materials they have been in
contact with, or contaminated needles, or
by aerosol dispersion.
Biological safety (airborne pathogens)
- The potential also exists for inadvertent
exposure to the public through direct
contact with aerosolized infectious
materials, improperly processed blood
products, and inappropriately disposed of
waste products.
Biological safety (general considerations)
- All blood samples and other body fluids
should be collected, transported, handled,
and processed using universal precautions
- Gloves, gowns, and face protection must be
BIOSAFETY CABINETS
used during manipulations or transfers
when splashing or splattering is most likely
to occur.
- Consistent and thorough hand washing is
an essential component of infection control.
Antiseptic gels and foams may be used at
waterless stations between washes.
- Centrifugation of biologic specimens
produces finely dispersed aerosols that are
a high-risk source of infection
Biological Safety (spills)
Mechanical hazard and safety
- o Alert others in area of the spill. o Wear
appropriate protective equipment.
- Use mechanical devices to pick up broken
glass or other sharp objects. o Absorb the
spill with paper towels, gauze pads, or
tissue. o Clean the spill site using a common
aqueous detergent.
- Disinfect the spill site using approved
disinfectant or 10% bleach, using
appropriate contact time.
- Rinse the spill site with water.
- Dispose of all materials in appropriate
biohazard containers.
- The CDC guidelines require the
development of a tuberculosis infection
control program by any facility involved in
the diagnosis or treatment of cases of
confirmed infectious TB. TB isolation areas
with specific ventilation controls must be
established in health-care facilities. Those
workers in high-risk areas may be required
to wear a respirator for protection. All
health-care workers considered to be at risk
must be screened for TB infection.
- Biological safety cabinets (BSCs) remove
particles that may be harmful to the
employee who is working with potentially
infectious biologic specimens.
- The Centers for Disease Control and
Prevention (CDC) and the National Institutes
of Health have described four levels of
biosafety, which consist of combinations of
laboratory practices and techniques, safety
equipment, and laboratory facilities.
- Centrifuges, for example, must be balanced
to distribute the load equally. The operator
should never open the lid until the rotor has
come to a complete stop. Safety interlocks
on equipment should never be rendered
inoperable
- Tongs or insulated gloves should be used to
remove hot glassware from ovens, hot
plates, or water baths.
- Glass pipets should be handled with extra
care, as should sharp instruments such as
cork borers, needles, scalpel blades, and
other tools. Such as cork borers, needles,
scalpel blades, and other tools.
Chemical hazard (flammable/combustible
chemicals)
- Flammable and combustible liquids, which
are used in numerous routine procedures,
are among the most hazardous materials in
the clinical chemistry laboratory because of
possible fire or explosion.
- A flammable liquid has a flash point below
37.8°C (100°F) and combustible liquids, by
definition, have a flash point at or above
37.8°C (100°F).
- Some commonly used flammable and
combustible solvents are acetone, benzene,
ethanol, heptane, isopropanol, methanol,
toluene, and xylene.
Chemical Hazard (corrosive chemical)
- Corrosive chemicals are injurious to the skin
or eyes by direct contact or to the tissue of
the respiratory and gastrointestinal tracts if
inhaled or ingested.
- Typical examples include acids (acetic,
sulfuric, nitric, and hydrochloric) and bases
(ammonium hydroxide, potassium
hydroxide, and sodium hydroxide).
Chemical hazard (reactive chemicals)
- Reactive chemicals are 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 reactions).
Hydrogen is liberated if alkali metals
(sodium or potassium) are mixed with water
or acids, and spontaneous combustion also
may occur.
- The mixture of oxidizing agents, such as
peroxides, and reducing agents, such as
hydrogen, generates heat and may be
explosive.
Chemical hazard (carcinogenic chemicals)
- Carcinogens are substances that have been
determined to be cancer-causing agents.
- Benzidine is a common example of a known
carcinogen.
Fire hazard
- Fire is basically a chemical reaction that
involves the rapid oxidation of a
combustible material or fuel, with the
subsequent liberation of heat and light. In
the clinical chemistry laboratory, all the
elements essential for fire to begin are
present—fuel, heat or ignition source, and
oxygen (air).
Electrical hazard
- Most individuals are aware of the potential
- Although increased mechanization and
hazards associated with the use of electrical
appliances and equipment. Direct hazards
of electrical energy can 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:
• Use only explosion-rated (intrinsically
wired) equipment in hazardous
atmospheres.
• Be particularly careful when operating
high-voltage equipment, such as
electrophoresis apparatus.
• Use only properly grounded equipment
(three-prong plug).
• Check for frayed electrical cords.
• Promptly report any malfunctions or
equipment producing a “tingle” for repair.
• Do not work on “live” electrical
equipment.
• Never operate electrical equipment with
wet hands.
• Know the exact location of the electrical
control panel for the electricity to your
work area.
• Use only approved extension cords in
temporary applications and do not overload
circuits. (Some local regulations prohibit the
use of any extension cord.)
• Have ground, polarity, and leakage checks
and other periodic preventive maintenance
performed on outlets and equipment.
Ergonomic Hazard
automation have made many tedious and
 repetitive manual tasks obsolete, laboratory
processes often require repeated
manipulation of instruments, containers,
and equipment. These physical actions can,
over time, contribute to repetitive strain
disorders such as tenosynovitis, bursitis,
and ganglion cysts. The primary
contributing factors associated with
repetitive strain disorders are
position/posture, applied force, and
frequency of repetition.
- Remember to consider the design of hand
tools (e.g., ergonomic pipettes), adherence
to ergonomically correct technique, and
equipment positioning when engaging in
any repetitive task. Chronic symptoms of
pain, numbness, or tingling in extremities
may indicate the onset of repetitive strain
disorders. Other hazards include acute
musculoskeletal injury. Remember to lift
heavy objects properly, keeping the load
close to the body and using the muscles of
the legs rather than the back. Gradually
increase force when pushing or pulling, and
avoid pounding actions with the
extremities.
Radiation Hazard
Environmental Protection
- A radiation safety policy should include
environmental and personnel protection. All
areas where radioactive materials are used
or stored must be posted with caution
signs, and traffic in these areas should be
restricted to essential personnel only.
Regular and systematic monitoring must be
emphasized, and decontamination of
laboratory equipment, glassware, and work
areas should be scheduled as part of
routine procedures. Records must be
maintained as to the quantity of radioactive
material on hand as well as the quantity
that is disposed.
Personal Protection
- It is essential that only properly trained
personnel work with radioisotopes. Good
work practices must consistently be
employed to ensure that contamination and
inadvertent internalization are avoided.
Users should be monitored to ensure that
the maximal permissible dose of radiation is
not exceeded. Radiation monitors must be
evaluated regularly to detect degree of
exposure for the laboratory employee.
Records must be maintained for the length
of employment plus 30 years.
Nonionizing Radiation
- Nonionizing forms of radiation are also a
concern in the clinical laboratory.
Equipment often emits a variety of
wavelengths of electromagnetic radiation
that must be protected against through
engineered shielding or use of PPE. These
energies have varying biologic effects,
depending on wavelength, power intensity,
and duration of exposure. Laboratorians
must be knowledgeable regarding the
hazards presented by their equipment to
protect themselves and ancillary personnel.