QUALITY CONTROL

Judy Anne B. Libu, RMT

CONTENTS
• QUALITY CONTROL
• CALCULATIONS AND USED OF QC STATISTICS
• LEVEY-JENNINGS CHART AND WESTGARD RULES
QUALITY CONTROL
• Quality control in the medical laboratory is a statistical process used to
monitor and evaluate the analytical process that produces patient results.
– REQUIREMENTS FOR A STATISTICAL PROCESS:
A. Regular testing of quality control products along with patient samples.
B. Comparison of quality control results to specific statistical limits (ranges).
QUALITY CONTROL
• When a diagnostic test is performed in the medical laboratory, the outcome
of the test is a result. The result may be a patient result or it may be a
quality control (QC) result. The result may be quantitative (a number) or
qualitative (positive or negative) or semi-quantitative (limited to a few
different values).
REGULAR TESTING OF QC
• Good laboratory practice requires testing normal and abnormal controls for
each test at least daily to monitor the analytical process. If the test is stable
for less than 24 hours or some change has occurred which could potentially
affect the test stability, controls should be assayed more frequently.
CALCULATION AND USED OF QC STATISTICS
CALCULATION AND USED OF QC STATISTICS
MEAN
• The mean (or average) is the laboratory’s best estimate of the analyte’s
true value for a specific
evel of control.
STANDARD DEVIATION
• Standard deviation is a statistic that quantifies how close numerical values
are in relation to each other. The term precision is often used
interchangeably with standard deviation. Another term, imprecision, is used
to express how far apart numerical values are from each other.
• It provides the laboratory an estimate of test consistency at specific
concentrations.
• The repeatability of a test may be consistent (low standard deviation, low
imprecision) or inconsistent (high standard deviation, high imprecision).
STANDARD DEVIATION
• IN CASES OF INSTABILITY OF THE RESULTS, THE FOLLOWING QUESTIONS
ARE USED TO INVESTIGATE THE PROBLEM
• Has the reagent or reagent lot changed recently?
• Has maintenance been performed routinely and on schedule?
• Does the potassium electrode require cleaning or replacement?
• Are the reagent and sample pipettes operating correctly?
• Has the test operator changed recently?
STANDARD DEVIATION
LEVEY-JENNINGS AND WESTGARD RULE
The Levey-Jennings chart
Horizontal graph of quality control data
• X-axis of the normal curve is converted to the vertical axis
○ Median flanked by 3 positive standard deviation increments above and 3
negative standard deviation increments below
○ Data points for each day plotted horizontally
○ Once the mean and standard deviations for an assay are determined, the
Levey-Jennings chart provides an easy -to-read visual representation of
quality control data generated over time
CREATING A LEVEY-JENNINGS CHART
• Standard deviation is commonly used for preparing Levey -Jennings (L-J or
LJ) charts. The Levey-Jennings chart is used to graph successive (run-to-run
or day-to-day) quality control values. A chart is created for each test and
level of control.
Normal Distribution
• In any large sample of random data, if the data points on the x-axis are
plotted against frequency on the y-axis, a bell-shaped curve will form, with
its peak coinciding on the x-axis with the mean of the sample
Normal (Gaussian) distribution
• Curve is symmetrical around the vertical line that passes through the
mean on the x-axis, and the curve approaches but never touches the x-axis
—is considered a normal, or Gaussian, distribution.
• The area under the curve represents the sum of all the frequencies of the
numbers in the sample, and the total sum is 1
• The normal curve is useful in the clinical laboratory when the units on the
x-axis are SD on either side of the mean
• In a normal curve
• 68% of all numbers will fall between - 1 SD and +1SD of the mean
• 95% will fall between - 2 SD and +2 SD
• 99% will fall between - 3 SD and +3 SD
• The values that fall within -/+ 2 SD of the mean are considered normal
and within the 95% confidence interval
• The values outside -/+ 2 SD but within -/+ 3 SD of the mean are
questionable
Systematic Error
• Systematic error is evidenced by a change in the mean of the control
values. The change in the mean may be gradual and demonstrated as a
trend in control values or it may be abrupt and demonstrated as a shift in
control values.
Trend
Six or more consecutive control values get continuously larger or smaller
(i.e., a linear direction of values)
○ Can move away from, toward, or across the mean
○ Represents a progressively worsening problem
A trend indicates a gradual loss of reliability in the test system. Trends are
usually subtle. Causes of trending may include:
– Deterioration of the instrument light source
– Gradual accumulation of debris in sample/reagent tubing
– Gradual accumulation of debris on electrode surfaces
– Aging of reagents
– Gradual deterioration of control materials
– Gradual deterioration of incubation chamber temperature (enzymes only)
– Gradual deterioration of light filter integrity
– Gradual deterioration of calibration
Trend
Six or more consecutive control values get continuously larger or smaller
(i.e., a linear direction of values)
○ Can move away from, toward, or across the mean
○ Represents a progressively worsening problem
A trend indicates a gradual loss of reliability in the test system. Trends are
usually subtle. Causes of trending may include:
– Deterioration of the instrument light source
– Gradual accumulation of debris in sample/reagent tubing
– Gradual accumulation of debris on electrode surfaces
– Aging of reagents
– Gradual deterioration of control materials
– Gradual deterioration of incubation chamber temperature (enzymes only)
– Gradual deterioration of light filter integrity
– Gradual deterioration of calibration
Shift
• Six or more control values fall on the same side of the mean
○ The distance from the mean does not matter
○ The problem does not necessarily grow steadily worse
○ A shift represents that a new mean has been established
• Abrupt changes in the control mean are defined as shifts. Shifts in QC data
represent a sudden and dramatic positive or negative change in test system
performance. Shifts may be caused by:
– Sudden failure or change in the light source
– Change in reagent formulation
– Change of reagent lot
– Major instrument maintenance
– Sudden change in incubation temperature (enzymes only)
– Change in room temperature or humidity
– Failure in the sampling system
– Failure in reagent dispense system
– Inaccurate calibration/recalibration
Random Error
• Technically, random error is any deviation away from an expected result.
For QC results, any positive or negative deviation away from the calculated
mean is defined as random error.
• There is acceptable (or expected) random error as defined and quantified
by standard deviation. There is unacceptable (unexpected) random error
that is any data point outside the expected population of data (e.g., a data
point outside the ±3s limits).
Westgard Rules
• In 1981, Dr. James Westgard of the University of Wisconsin published an
article on laboratory quality
control that set the basis for evaluating analytical run quality for medical
laboratories. The elements of the Westgard system are based on principles
of statistical process control used in industry nationwide since the 1950s.
• There are six basic rules in the Westgard scheme. These rules are used
individually or in combination to evaluate the quality of analytical runs.
Westgard Multirule set
• Monitor quality control performance is based on control rules.
• Whether violation of these rules constitutes a warning or a rejection
depends on individual laboratory policies
• 12s: One control exceeds -/+ 2 SD of the mean (day 1, level 1)
• This is a warning rule that is violated when a single control observation is
outside the ±2s limits. Remember that in the absence of added analytical
error, about 4.5% of all quality control results will fall
between the 2s and 3s limits. This rule merely warns that random error or
systematic error may be present in the test system.
The relationship between this value and other control results within the
current and previous analytical runs must be examined. If no relationship
can be found and no source of error can be identified, it must be assumed
that a single control value outside the ±2s limits is an acceptable random
error. Patient results can be reported.
• 13s: One control exceeds - /+ 3 SD of the mean; indicative of random
error
(day 2, level 1)
• This rule identifies unacceptable random error or possibly the beginning of
a large systematic error. Any QC result outside ±3s violates this
• 22s: Two consecutive controls are either +2 SD or -2 SD of the mean;
indicative of systematic error (day 4, levels 1 and 2; or day 4 and 5, level 1)
• This rule identifies systematic error only.
The criteria for violation of this rule are:
– Two consecutive QC results
– Greater than 2s
– On the same side of the mean
• R4s: Combination of one control +2 SD of the mean and another -2 SD of
the mean; indicative of random error (day 6, levels 1 and 2)
• This rule identifies random error only, and is applied only within the current
run. If there is at least a 4s difference between control values within a single
run, the rule is violated for random error.
For example, assume both Level I and
Level II have been assayed within the
current run. Level I is +2.8s above the
mean and Level II is -1.3s below the
mean. The total difference betweenthe
two control levels is greater than 4s (e.g.
[+2.8s – (-1.3s)] = 4.1s).
• Violation of any of the following rules does not necessarily require rejection
of the analytical run. These violations typically identify smaller systematic
error or analytical bias that is not often clinically significant or relevant.
• Analytical bias may be eliminated by performing calibration or instrument
maintenance.
• 31s:The criteria which must be met to violate this rule are:
– Three consecutive results
– Greater than 1s
– On the same side of the mean
• 41s :The criteria which must be met to violate this rule are:
– Four consecutive results
– Greater than 1s
– On the same side of the mean
• 10x: 10 consecutive controls falling on one side or the other of the mean;
indicative of systematic error
• These rules are violated when there are:
– 7 or 8, or 9, or 10, or 12 control results
– On the same side of the mean regardless of the specific standard deviation
in which they are located.

CLASSICAL METHODS OF ANALYSIS

CLASSICAL METHODS OF ANALYSIS
• Classical methods/ wet chemical methods/ earliest methods of analysis-
Relied mainly on chemical properties of analytes.
• Analytes are treated with reagents to form products that could be
identified.
• Gravimetric and titrimetric methods were used for quantitative analysis.
CLASSICAL METHODS OF ANALYSIS
Examples:
1.Formation of precipitate and measurement of mass.
2.Oxidation of analyte and detection of end via the change of color of the
analyte.
3.Neutralization of analyte and detection of end point using acidbase
indicator.
4.Complexation of analyte and use of metallochromic indicators to detect
end point.
Separation of analyte from matrix was achieved using precipitation,
extraction and distillation.
GRAVIMETRIC METHOD CLASSICAL METHODS OF ANALYSIS
GRAVIMETRIC METHOD
• Gravimetric analysis describes as a set of methods used in analytical
chemistry for the quantitative determination of an analyte (the ion
being analyzed) based on its mass.
• The principle behind this type of analysis is that once an ion's mass
has been determined as a unique compound, that known
measurement can then be used to determine the same analyte's
mass in a mixture, as long as the relative quantities of the other
constituents are known.
GRAVIMETRIC METHOD
GRAVIMETRIC METHOD
ADVANTAGES:
• Provides for exceedingly precise analysis
• Gravimetric analysis was used to determine the atomic masses of many
elements to six figure accuracy.
• Gravimetry provides very little room for instrumental error
• It does not require a series of standards for calculation of an unknown.
• Also, methods often do not require expensive equipment.
• Gravimetric analysis, due to its high degree of accuracy, when performed
correctly, can also be used to calibrate other instruments in lieu of reference
standards.
GRAVIMETRIC METHOD
DISADVANTAGES:
• Gravimetric analysis usually only provides for the analysis of a single
element, or a limited group of elements, at a time.
• Comparing modern dynamic flash combustion coupled with gas
chromatography with traditional combustion analysis will show that
the former is both faster and allows for simultaneous determination
of multiple elements while traditional determination allowed only for
the determination of carbon and hydrogen.
• Methods are often convoluted and a slight mis-step in a procedure
can often mean disaster for the analysis (colloid formation in
precipitation gravimetry, for example).
GRAVIMETRIC METHOD
DISADVANTAGES:
• Gravimetric analysis usually only provides for the analysis of a single
element, or a limited group of elements, at a time.
• Comparing modern dynamic flash combustion coupled with gas
chromatography with traditional combustion analysis will show that
the former is both faster and allows for simultaneous determination
of multiple elements while traditional determination allowed only for
the determination of carbon and hydrogen.
• Methods are often convoluted and a slight mis-step in a procedure
can often mean disaster for the analysis (colloid formation in
precipitation gravimetry, for example).
GRAVIMETRIC METHOD
•FOUR MAIN TYPES OF GRAVIMETRIC METHOD:
1. Precipitation
2. Volatilization
3. Electro-analytical
4. Miscellaneous physical method
a. Gravimetric titrimetry
b. Atomic Mass Spectroscopy
PRECIPITATION
GRAVIMETRY
• In precipitation gravimetry, the analyte is separated from a solution of the
sample as a precipitate and is converted to a compound of known
composition that can be weighed.
• The analyte used in precipitation gravimetry is converted into a sparingly
soluble precipitate. The precipitate is filtered, washed free of impurities,
converted to a product of known composition by suitable heat treatment and
weighed.
PRECIPITATION
GRAVIMETRY
PROCEDURE:
1. PRECIPITATION and DIGESTING
2. FILTERING THE PRECIPITATE
3. RINSING THE PRECIPITATE
4. DRYING AND WEIGHING THE FINAL
PRECIPITATE
PRECIPITATION GRAVIMETRY
1. Precipitating and Digesting:
Properties of Precipitates and
Precipitating Reagents:
1.Easily filtered and washed free contaminants
2.Sufficiently low solubility that no significant loss of the analyte occurs
during filtration and washing
3.Unreactive with constituents of the atmosphere
4.Known chemical coposition after it is dried or, if necessary, ignited.
PRECIPITATION
GRAVIMETRY
AVOIDING IMPURITIES:
• In addition to having a low solubility, the precipitate must be free from
impurities. Because precipitation usually occurs in a solution that is rich in
dissolved solids, the initial precipitate is often impure. We must remove
these impurities before determining the precipitate’s mass.
• The greatest source of impurities is the result of chemical and physical
interactions occurring at the precipitate’s surface. A precipitate is generally
crystalline—even if only on a microscopic scale—with a well-defined lattice of
cations and anions.
PRECIPITATION
GRAVIMETRY
CONTROLLING THE PARTICLE SIZE:
A homogeneous precipitation produces large particles of precipitate that are
relatively free from impurities. These advantages, however, are offset by
requiring more time to produce the precipitate and a tendency for the
precipitate to deposit as a thin film on the container’s walls. The latter
problem is particularly severe for hydroxide precipitates generated using
urea.
PRECIPITATION
GRAVIMETRY
• A visible precipitate takes longer to form when RSS is small both
because there is a slow rate of nucleation and because there is a
steady decrease in RSS as the precipitate forms. One solution to the
latter problem is to generate the precipitant in situ as the product of
a slow chemical reaction. This maintains the RSS at an effectively
constant level. Because the precipitate forms under conditions of low
RSS, initial nucleation produces a small number of particles. As
additional precipitant forms, particle growth supersedes nucleation,
resulting in larger precipitate particles. This process is called
homogeneous precipitation.
PRECIPITATION
GRAVIMETRY
Two precipitates of PbCrO4. In Beaker A, combining 0.1 M Pb(NO3)2 and 0.1
M
K2CrO4 forms the precipitate under conditions of high RSS. The precipitate
forms rapidly and consists of very small
particles. In Beaker B, heating a solution
of 0.1 M Pb(NO3)2, 0.1 M Cr(NO3)3, and
0.1 M KBrO3 slowly oxidizes Cr3+ to
CrO42–, precipitating PbCrO4 under
conditions of low RSS. The precipitate
forms slowly and consists of much larger
particles.
PRECIPITATION
GRAVIMETRY
Two precipitates of PbCrO4. In Beaker A,
combining 0.1 M Pb(NO3)2 and 0.1 M
K2CrO4 forms the precipitate under
conditions of high RSS. The precipitate
forms rapidly and consists of very small
particles. In Beaker B, heating a solution
of 0.1 M Pb(NO3)2, 0.1 M Cr(NO3)3, and
0.1 M KBrO3 slowly oxidizes Cr3+ to
CrO42–, precipitating PbCrO4 under
conditions of low RSS. The precipitate
forms slowly and consists of much larger
particles.
PRECIPITATION
GRAVIMETRY
2. FILTERING THE PRECIPITATE
After precipitating and digesting the precipitate, we separate
it from solution by filtering. The most common filtration
method uses filter paper, which is classified according to its
speed, its size, and its ash content on ignition. Speed, or how
quickly the supernatant passes through the filter paper, is a
function of the paper’s pore size. A larger pore allows the
supernatant to pass more quickly through the filter paper, but
does not retain small particles of precipitate.
PRECIPITATION GRAVIMETRY
Proper procedure for transferring the supernatant to the filter paper cone.
PRECIPITATION
GRAVIMETRYAn
alternative method for filtering a precipitate is a filtering crucible. The trap
prevents water from an aspirator from back-washing into the suction flask.
PRECIPITATION
GRAVIMETRY3. RINSING THE PRECIPITATE
Because the supernatant is rich with dissolved inert ions, we must
remove any residual traces of supernatant to avoid a positive
determinate error without incurring solubility losses.
In many cases this simply involves the use of cold solvents or rinse
solutions containing organic solvents such as ethanol. The pH of the
rinse solution is critical if the precipitate contains an acidic or basic
ion.
Adding a volatile inert electrolyte to the rinse solution prevents the
precipitate from reverting into smaller particles that might pass
through the filter. This process of reverting to smaller particles is
called peptization. The volatile electrolyte is removed when drying
the precipitate.
PRECIPITATION
GRAVIMETRY
4. DRYING THE PRECIPITATE
After separating the precipitate from its supernatant solution, the
precipitate is dried to remove residual traces of rinse solution and any
volatile impurities.
The temperature and method of drying depend on the method of
filtration and the precipitate’s desired chemical form. Placing the
precipitate in a laboratory oven and heating to a temperature of 110°C
is sufficient when removing water and other easily volatilized
impurities.
Higher temperatures require a muffle furnace, a Bunsen burner, or a
Meker burner, and are necessary if we need to thermally decompose
the precipitate before weighing.
VOLATILIZATION
GRAVIMETRY• In volatilization gravimetry, the analyte is separated from
other
constituents of a sample by converting it to a gas known
chemical composition. The mas of the gas then serves as a
measure of the analyte concentration.
• In volatilization methods, removal of the analyte involves
separation by heating or chemically decomposing a volatile
sample at a suitable temperature In other words, thermal or
chemical energy is used to precipitate a volatile species For
example, the water content of a compound can be determined
by vaporizing the water using thermal energy (heat). Heat can
also be used, if oxygen is present, for combustion to isolate the
suspect species and obtain the desired results.
VOLATILIZATION
GRAVIMETRY
One method for determining the products of a
thermal decomposition is to monitor the sample’s
mass as a function of temperature, a process
called thermogravimetry.
VOLATILIZATION
GRAVIMETRY
• Depending on the method of analysis, the equipment for volatilization
gravimetry may be simple or complex. In the simplest experimental design,
we place the sample in a crucible and decompose it at a fixed temperature
using a Bunsen burner, a Meker burner, a laboratory oven, or a muffle
furnace. The sample’s mass and the mass of the residue are measured using
an analytical balance.
• Trapping and weighing the volatile products of a thermal decomposition
requires specialized equipment. The sample is placed in a closed container
and heated. As decomposition occurs, a stream of an inert purge-gas sweeps
the volatile products through one or more selective absorbent traps.
VOLATILIZATION
GRAVIMETRY• In a thermogravimetric analysis, the sample is placed on a
small
balance pan attached to one arm of an electromagnetic balance. The
sample is lowered into an electric furnace and the furnace’s
temperature is increased at a fixed rate of few degrees per minute
while continuously monitoring the sample’s weight. The instrument
usually includes a gas line for purging the volatile decomposition
products out of the furnace, and a heat exchanger to dissipate the
heat emitted by the furnace.
VOLATILIZATION
GRAVIMETRY
VOLATILIZATION
GRAVIMETRY
VOLATILIZATION
GRAVIMETRY• Volatilization methods can be either direct or indirect. Water
eliminated in a
quantitative manner from many inorganic substances by ignition is an
example of a direct determination. It is collected on a solid desiccant and its
mass determined by the gain in mass of the desiccant.
• Another direct volatilization method involves carbonates which generally
decompose to release carbon dioxide when acids are used. Because carbon
dioxide is easily evolved when heat is applied, its mass is directly established
by the measured increase in the mass of the absorbent solid used
• Determination of the amount of water by measuring the loss in mass of the
sample during heating is an example of an indirect method. It is well known
that changes in mass occur due to decomposition of many substances when
heat is applied, regardless of the presence or absence of water. Because one
must make the assumption that water was the only component lost, this
method is less satisfactory than direct methods.
ELECTROGRAVIMETRY
• In electrogravimetry, the analyte is separated by deposition on an
electrode by an electrical current. The mass of this product then
provides a measure of the analyte concentration.
• Electrolytic deposition has been used for over a century for the
gravimetric determination of metals. In most applications, the metal
is deposited on a weighed platinum cathode, and the increase in
mass is determined. Some methods use anodic deposition such as the
determination of lead as lead dioxide on platinum and of chloride as
silver or chloride on silver.
• Working electrode- is the electrode in which the analytical reaction
occurs.
General types of Electrogravimetric
Methods:
1.Uncontrolled Potential of the working electrode
Electrolytic procedures in which the potential of the working electrode is
uncontrolled use simple and inexpensive equipment and require a little
operator attention. In these procedures, the potential applied across the
entire
cell is maintained at a more-or-less constant level throughout the
electrolysis.
2.Controlled-Potential/ Potentiostatic method
The controlled-potential apparatus is made up of two independent electrical
circuits that share a common electrode, the working electrode where the
analyte is deposited. The electrolysis current passes between the working
electrode and a counter electrode. The counter electrode has no effect on
the
reaction at the working electrode. A potentiostat maintains the working
electrode potential at a constant value relative to a reference electrode.
COULOMETRIC METHOD
The quantity of electrical charge required to convert a sample of an
analyte quantitatively to a different oxidation state is measured.
Coulometric and gravimetric share the common advantage that the
proportionality constant between the quantity measured and the
analyte mass is calculated from accurately known physical constants,
which can eliminate the need for calibration with cheical standards.
In contrast to gravimetric methods, coulometric procedures are usually
rapid and do not require that the product of the electrochemical
reaction be a weighable solid.
Coulometric methods are as accurate as conventional gravimetric and
volumetric procedures an in addition are easily automated.
GRAVIMETRIC
TITRIMETRY
In gravimetric titrimetry, the mass of a reagent of known concentration
required to react completely with the analyte provides the information
needed to determine the analyte concentration. In a mass/ gravimetric
titrations, a balnce and a weighable solution dispenser are substituted for a
buret and its markings.
Advantages of Gravimetric
Titrations:
1. Greater speed and convenience
2. Calibration of glasswares and tedious cleaning to ensure proper drainage
are completely eliminated.
3. Temperature corrections are unnecessary because the mass (weight)
molar concentrations does not change with
temperature, in contrast to the molar volume concentration.
4. Mass measurements can be made with considerably
greater precision and accuracy that can volume
measurements.
5. Gravimetric titrations are more easily automated than are
ATOMIC MASS
SPECTROMETRY
Atomic mass spectrometry uses a mass spectrometer to separate the
gaseous ions formed from the elements making up a sample of matter. The
concentration of the resulting ions is then determined by measuring the
electrical current produced when they fall on the surface of an ion detector.
Atomic mass spectrometry is a quantitative tool that can determine nearly
all the elements in the periodic table. Detection limits are often several
orders of magnitude better than optical methods.
TITRIMETRIC
METHODS
CLASSICAL METHODS OF ANALYSIS
TITRIMETRIC
METHODS
Titration methods, often called titrimetric methods,include a large and
powerful group of quantitative procedures based on measuring the amount
of a reagent of unknown concentration that is consumed byan analyte in a
chemical or electrochemical reaction. The reagent may be a standard
solution of a chemical or an electric current of known magnitude.
TITRIMETRIC
METHODSTitration methods, often called titrimetric methods, include a large
and powerful group of quantitative procedures based on measuring the
amount of a reagent of unknown concentration that is consumed byan
analyte in a chemical or electrochemical reaction. The reagent may be a
standard solution of a chemical or an electric current of known magnitude.
The three types of titrimetric methods
include:
1. Volumetric methods - measuring the volume of a solution
(of known concentration) required to react completely with
an analyte.
2. Gravimetric titrimetry - measuring the mass of a known reagent required
to react completely with an analyte.
3. Coulometric titrimetry - measuring the electrical current (amps) produced
when a known amount of reagent reacts completely with an analyte.
FLUOREMETRY
FLUOREMETRY
Fluorescence in an energy emission that occurs when a certain
compounds absorb electromagnetic radiation become excited and
then return to an energy level that is usually slightly higher than their
original level.
Fluorescence concentration is related to:
1.Molar absorbtivity of the compound
2.Intensity of radiation
3.Quantum efficiency
4.Length of light path
FLUOROMETRY
Phosphorescence- luminescence that is much longer. It is the emitted energy
equated to or lower than the absorbed energy.

Chemiluminescence method- produced when a chemical reaction yields an

electronically excited molecule, which emits light as it returns to the ground
state. Chemiluminescence methods are known for their high sensitivities.
TURBIDIMETRY
TURBIDIMETRY
TURBIDIMETRY
Turbidimetry is the measurement of the light blocked by a suspension of
particulate matter as light passes through the cuvette.
Factors affecting Turbidimetry:
1.Size and number of particles
2.The depth of the tube
3.Cross-sectional area of each particle
NEPHELOMETRY
NEPHELOMETRY
Nephelometry is the measurement of light scattered by small particles at an
angle to the beam incident on the cuvette. It is more specific than
Turbidimetry.
MORE ON TITRIMETRIC ANALYSIS
TITRIMETRIC ANALYSIS
• Titrimetric analysis, formerly known as the volumetric analysis, refers to
quantitative chemical analysis carried out by determining the volume of a
solution of accurate known concentration which is required to react
quantitatively with the solution of the substance to be determined.
DEFINITION OF TERMS
• STANDARD SOLUTION is a solution of accurate known strength
– The weight of the substance to be determined is calculated from the
volume of the standard solution used.
• The process of adding the standard solution until the reaction just
complete is termed as TITRATION and the substances to be determined is
TITRANT.
• INDICATOR is known as the auxillary m. It gives a clear visual change
(either a change in color or the formation of turbidity) in the solution being
titrated.
END POINT is the point at which the visual change of a solution occurs.
Ideal titration:
1. There must be a simple reaction which can be expressed by a chemical
equation, the substance to be determined should react completely with the
reagent in stoichiometric or equivalent portion.
2. There must be marked change in the free energy to alter chemical or
physical properties of the solution at the equivalence point.
3. The reaction should practically instantaneous or rapid. In some cases, a
catalyst is added to increase the speed of the reaction.
4. An indicator should be available which should sharply define the end pint
of the reaction.
Types of Titrimetric Analysis:
1. Neutralization Reactions (Acidemetry and Alkalimetry)
2. Complex Formation Reactions
3. Precipitation Reactions
4. Oxidation-Reduction Reactions
Neutralization Reactions (Acidemetry and Alkalimetry)
• These include the titration of bases with a standard acid (alkalimetry)
and the titration of acids with a standard base (acidemetry).
• It involves the combination of hydronium (H3O+) and hydroxyl ions (OH-)
to form water.
Complex Formation Reactions
• These depend upon the combination of ions, other than hydronium and
hydroxyl ions, to form soluble, slightly dissociated ions or compound, as in
the titration of metal ions with an organic complexion agent such as ethylene
diaminetetra acetic acid (EDTA).
Complex Formation Reactions
• These depend upon the combination of ions, other than hydronium and
hydroxyl ions, to form soluble, slightly dissociated ions or compound, as in
the titration of metal ions with an organic complexion agent such as ethylene
diaminetetra acetic acid (EDTA).
Precipitation Reactions
• These depend upon the combination of ions to form a simple precipitate
as the titration of silver ion with a solution of chloride ion.
Oxidation-Reduction Reactions
• These includes all reactions involving change in oxidation number or
transfer of electrons among the reacting substances.
• The standard solutions are either oxidizing or reducing agents.
Oxidation-Reduction Reactions
• Common oxidizing
agents:
1.KMnO4
2.K2Cr2O7
3.Ce(SO2)2
4.I2
5.KIO3
6.KBrO3
• Frequently used
reducing agents
1.Fe2+
2.Sn2+
• Compounds
1.Na2S2O3
2.As2O3
3.HgNO3
4.CrCl2
5.TiCl3 or Ti2(SO4)3
Oxidation-Reduction Reactions
• Common oxidizing
agents:
1.KMnO4
2.K2Cr2O7
3.Ce(SO2)2
4.I2
5.KIO3
6.KBrO3
• Frequently used
reducing agents
1.Fe2+
2.Sn2+
• Compounds
1.Na2S2O3
2.As2O3
3.HgNO3
4.CrCl2
5.TiCl3 or Ti2(SO4)3
Standard Solutions
• Contains a known weight of the reagent in a definite volume of solution,
concentrations are expressed in terms of molarity and normality.
• Normal Solution is a solution containing one equivalent of a defined species
per liter according to the specified reaction and a molar solution containing
one mole of a defined species per liter.
Preparation of Standard Solution
1. The preparation of standard solution can be made easier when the
material to be dissolved is of known purity.
2. A careful weighed or measured volume of sample is usually transfered to
a graduated flask and diluted to an exact volume.
3. It is undesirable to use solution of low concentration that one or two drops
of reagents are required to see a change in color of the indicator used.
4. The use of concentrated solution is also not feasible.
5. Commonly used are prepared solution of 0.1N concentration.
Preparation of Standard Solution
1. The preparation of standard solution can be made easier when the
material to be dissolved is of known purity.
2. A careful weighed or measured volume of sample is usually transfered to
a graduated flask and diluted to an exact volume.
3. It is undesirable to use solution of low concentration that one or two drops
of reagents are required to see a change in color of the indicator used.
4. The use of concentrated solution is also not feasible.
5. Commonly used are prepared solution of 0.1N concentration.
PREPARATION OF STANDARD SOLUTION
Standardization of solutions requires:
1. The primary standard must be readily available.
2. The weight and/or volume of the standard used should not be too small.
Never use a volume less than 10mL.
3. Back tirations during standardization must be avoided because the
process increases the possibility of errors.
4. Standardization must be performed in 3 trials and the results should
agree within 0.1-0.2%
Primary Standard Substances
• 1. Should be easily obtainable and easy to purify, to dry and to preserve in
a pure form.
• 2. It should remain stable I air during weighing, that is, it should be
nonhygroscopic (moist), not oxidized by air not affected by CO2.
• 3. It must be of definite, known composition.
• 4. It should be capable of being tested for impurities by qualitative and
other tests of known sensitivity. (the total amount of impurities should not in
general exceed 0.01- 0.02%)
• 5. It should have a high equivalent weight so that the weighing errors
may be negligible.
• 6. It should be readily soluble under the condition in which it is
employed.
Methods of Performing Titrations:
A. Direct Method
B. Indirect Method
C. Back Titration Method
D. Titer Method
Direct Method
• Dead-Stop End Point
• The amount of standard solution added is just up to end.
• This can be known
by the formula:
Indirect Method
• A measured amount of A is added in excess of that theoretically added to
react with B.
• Then the excess A is titrated to end point.
• This method can be expressed by the formula:
Back Titration Method
• When the end point is overstepped or an excess of the titrant has been
added unintentionally, the process of back titration is
carried out.
• The second titrant, which has the same nature as the analyte is used to
get back the end point of titration.
• The formula is:
Titer Method
• The weight of the sample equivalent to 1mL of a reagent is called
TITER.
• It is used to describe the strength of a solution in terms of anoter
reagent.
Acid-Base Titration/ Neutralization Reactions
• The equivalent of an acid is a mass of it which contains 1.008 (more
accurately 1.0078)of replaceable hydrogen.
• The equivalent of a monoprotic acid, such as HCl, HI, HClO4 of HOAc,
identical with the mole.
• A normal solution of a monoprotic acid will therefore contain one mole per
liter of solution.
• The equivalent of a diprotic acid (example: H2SO4 or H2C2O4) of triprotic
acid (Example H3PO4) is likewise 1/2 or 1/3 respectively of the mole.
• The equivalent of a base is that mass of it which contains one replaceable
hydroxyl group, that is 17.008g or ionizable; 17.008g hydroxyl are
equivalent to 1.008g hydrogen.
• The equivalents of NaOH and KOH are the mole of Ca(OH)2, Sr(OH)2, and
the Ba(OH)2 half a mole.
• Salt of strong bases and weak acids possess alkaline reaction in aqueous
solution because of
hydrolysis.
• A mole of Na2CO3 with methyl orange as indicator, reacts with two mole of
HCl hence is equivalent to half a mole.
• Sodium tetraborate under similar conditions, also reacts with two mole HCl
and its equivalent is likewise half a mole.
Preparation of Standard Acids and Bases
• Standard solutions of acids are usually prepared from HCl and H2SO4
• Both of these are commerically available as concentrated solutions,
concentrated HCl is about 10.5-12M and concentrated H2SO4 is about 18M.
• By suitable dilutions, solution of any desired approximate strength may
be readily prepared.
• HCl is generally preferred since most chlorides are water soluble.
• H2SO4 forms insoluble salts with calcium and barium hydroxides, for
titration of hot liquids or for determinations which requires boiling for
sometime with excess of acid, standard H2SO4 is, however preferable.
• NItric acid is seldom used, as standard it contains acid, which has a
destructive effect upon many indicators.
• The hydroxides of Sodium, Potassium and Barium are widely used for the
preparation of solution of standard alkalies, they are water soluble strong
bases.
• Solution prepared from aqueous ammonia are undesirable because they
tend to lose ammonia arise in titration with weak alkalies.
Primary Standard for Alkalies
• 1.Potassium hydrogen phthalate (KHC8H4O4/ KHP)- is an excellent
primary standard for bases. It has a high purity of atleast 99.95. it is almost
nonhygroscopic and has a high equivalent weight of 204 g/eq. It is
sufficiently stable to be dried at 120°C. Phenolphthalein indicator is
employed in the titration.
• 2.Benzoic acid (C6H6COOH)- it is a weak monobasic acid which is
sparingly soluble in water but soluble in a mixture of 95% ethyl alcohol and
water.
• 3.Succinic acid ((CH2COOh)2)- fairly soluble in water.
Primary Standard for Alkalies
• 4. Potassium Hydrogen Iodate (KH(IO3)O2)- it is a strong monoprotic acid
which permits the use of any indicator having a pH range between 4.5-9.5
for titration with strong bases. It is moderately soluble in water, is anhydrous
and non-hygroscopic and its aqueous solution is soluble for long periods.
• 5. Sulfamic acid (HSO3NH3)- it is a colorless cystalline, non- hygroscopic
solid, melting with decomposition at 205°C. the acid is moderately soluble in
water. Sulfamic acid acts as a strong acid, so that any indicator with a color
change in the pH range 4- 9 may be employed.
Primary Standard for Acids
• 1. Sodium carbonate (Na2CO3)- it is commonly employed as a priary
standard for acid solution. It is obtained commericially in purity of 99.95%.
this contains a little moisture and must be dehydrated by heating at 260°C-
270°C. more commonly it is titrated using methyl orange indicator.
• 2. Tris (Hydroxymethyl) aminomethane ((C2HOH)3CNH2)- abbreviated as
TRIS or THAM is an excellent primary standard for solution of strong acids. It
is an organic base which is readily available in purity of 99.95%, soluble in
water, has a high equivalent weight, stable and does absorb CO.
• 3.Sodium tetraborate or borax (Na2B4O7)- has the following advantages:
it has a large equivalent weight, it is easily and economically purified by
crystallization; it is not hygrocopic and a sharp end point can be obtained
with methyl red.
Primary Standard for Acids
• 1. Sodium carbonate (Na2CO3)- it is commonly employed as a priary
standard for acid solution. It is obtained commericially in purity of 99.95%.
this contains a little moisture and must be dehydrated by heating at 260°C-
270°C. more commonly it is titrated using methyl orange indicator.
• 2. Tris (Hydroxymethyl) aminomethane ((C2HOH)3CNH2)- abbreviated as
TRIS or THAM is an excellent primary standard for solution of strong acids. It
is an organic base which is readily availabble in purity of 99.95%, soluble in
water, has a high equivalent weight, stable and does absorb CO.
• 3.Sodium tetraborate or borax (Na2B4O7)- has the following advantages:
it has a large equivalent weight, it is easily and economically purified by
crystallization; it is not hygrocopic and a sharp end point can be obtained
with methyl red.
Indicators for Acid-Base Titrations
• The object of titrating, like an alkaline solution with standard solutions of
an acid is the determination of the amount of base present.
• The point at which this is reached as the equivalence point; an aqueous
solutions of the corresponding salt results.
• A large number of substances available are called neutralization or acid-
base indicators, which possess different colors according to the hydrogen ion
concentration of the solution.
• Indicators are organic compounds which may either be weak acids or weak
bases that can change color at a define pH range