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ANALYTICAL CHEMISTRY
Chapter 1
Introduction
Analytical Chemistry deals with methods
for determining the chemical composition of
samples.
• Qualitative Analysis (identification)
provides information about the identity of
species or functional groups in the
sample (an analyte can be identified).
• Quantitative Analysis provides numerical
information of analyte (quantitate the
exact amount or concentration).
Analytical Methods
• Classical Methods: Wet chemical
methods such as precipitation, extraction,
distillation, boiling or melting points,
gravimetric and titrimetric measurements.
• Instrumental Methods: Analytical
measurements (conductivity, electrode
potential, light absorption or emission,
masstocharge ratio, fluorescence etc.)
are made using instrumentation.
Types of Instrumental Methods
1. Spectroscopic methods:
a. Atomic spectroscopy
b. Molecular spectroscopy
2. Chromatographic methods
(separations):
3. Electrochemistry:
Block diagram of an instrumental measurement
Block diagram of a fluorometer
Applications of Instrumental
Methods
1. Bioanalytical: biological molecules and/or
biological matrices (e.g., proteins, amino
acids, blood, urine)
2. Environmental: pesticides, pollution, air,
water, soil
3. Material science: polymers,
characterization of new materials
4. Forensic science (application of science to
the law): body fluids, DNA, gun shot
residue, hair, fibers, elemental analysis,
drugs, alcohols, poisoning, fingerprints, etc.
Analytical Methodology
1. Plan: Qualitative or quantitative or both; what
kind of information have; which technique is
suitable etc.
2. Sampling: Accuracy depends on proper
sampling, characteristic of sample is very
important, required good representative
sample (from top, middle and bottom and mix
up and take average sample).
3. Sample preparation: depends on analytical
techniques.
4. Analytical measurement:
5. Data Analysis: Whether the data make sense
or not.
Selecting an Analytical Method
In order to select an analytical method intelligently, it is
essential to define clearly the nature of the analytical
problem. In general, the following points should be
considered when choosing an instrument for any
measurement.
1. Accuracy and precision required
2. Available sample amount
3. Concentration range of the analyte
4. Interference in sample
5. Physical and chemical properties of the sample matrix
6. Number of sample to be analyzed
7. Speed, ease, skill and cost of analysis
Figures of Merit
Precision
Bias
Sensitivity
Detection limit
Concentration range (Dynamic range)
Selectivity
Precision: How close the same measurements are
to one another. The degree of mutual agreement
among data that have been obtained in the same
way. Precision provides a measure of the random
or indeterminate error of an analysis.
Accuracy: How close the measurement
approaches the real value.
Bias: Bias provides a measure of the systematic,
or determinate error of an analytical method.
bias = µ  x
t
, where, µ is the population mean and
x
t
is the true value
Sensitivity: Sensitivity of an instrument is a
measure of its ability to discriminate between
small differences in analyte concentration. The
change in signal per unit change in analyte
concentration. The slope of the calibration curve at
the concentration of interest is known as
calibration sensitivity.
S = mc + S
bl
S = measured signal; c= analyte concentration;
S
bl
= blank signal; m = sensitivity (Slope of line)
Analytical sensitivity (¸)
¸ = m/s
s
m = slope of the calibration curve
s
s
= standard deviation of the measurement
Detection Limit (Limit of detection, LOD): The minimum
concentration of analyte that can be detected with a specific
method at a known confidence level.
LOD is determined by S/N, where, S/N = Signaltonoise ratio
= (magnitude of the signal)/(magnitude of the noise)
• Noise: Unwanted baseline fluctuations in the absence of
analyte signal (standard deviation of the background)
• The detection limit is given by,
C
m
= (S
m
– S
bl
)/m, where, C
m
= minimum concentration i.e.,
LOD, S
m
= minimum distinguishable analytical signal (i.e.,
S/N = 2 or S/N = 3), S
bl
= mean blank signal
m = sensitivity (i.e., slope of calibration curve)
• The amount of analyte necessary to yield a net signal equal
to 2 or 3x the standard deviation of the background.
Dynamic Range: The lowest concentration at which
quantitative measurements can be made (limit of quantitation,
or LOQ) to the concentration at which the calibration curve
departs from linearity (limit of linearity, or LOL).
The lower limit of quantitative measurements is generally
taken to be equal to ten times the standard deviation of
repetitive measurements on a blank or 10 S
bl
.
Dynamic range is the range over which detector still responds
to changing concentration (at high concentrations – usually
saturates – quits responding)
An analytical method should have a dynamic range of at least
two orders of magnitude, usually 26 orders of magnitude.
Selectivity: Selectivity of an analytical method
refers to the degree to which the method is free
from interference by other species contained in the
sample matrix. No analytical method is totally free
from interference from other species, and steps
need to be taken to minimize the effects of these
interferences. Selectivity coefficient gives the
relative response of the method to interfering
species as compared with analyte. Selectivity
coefficient can range from zero (no interference) to
values greater than unity. A coefficient is negative
when the interference caused a reduction in the
intensity of the output signal of the analyte.
Calibration of Instrumental
Methods
All types of analytical methods require calibration for
quantitation. Calibration is a process that relates the
measured analytical signal to the concentration of
analyte. We can’t just run a sample and know the
relationship between signal and concentration without
calibrating the response
The three most common calibration methods are:
• Calibration curve
• Standard addition method
• Internal standard method
Calibration Curves
• Several standards (with different concentration) containing exactly
known concentrations of the analyte are measured and the responses
recorded.
• A plot is constructed to give a graph of instrument signal versus analyte
concentration.
• Sample (containing unknown analyte concentration) is run, if response
is within the LDR of the calibration curve then concentration can be
quantitated.
• Calibration curve relies on accuracy of standard concentrations.
• It depends on how closely the matrix of the standards resemble that of
the sample to analyzed.
• If matrix interferences are low, calibration curve methods are OK.
• If matrices for sample and standards are not same calibration curve
methods are not good.
• Need to consider the linear part of the curves.
Standard Addition Methods
Better method to use when matrix effects can be substantial
Standards are added directly to aliquots of the sample,
therefore matrix components are the same.
Procedure:
• Obtain several aliquots of sample (all with the same
volume).
• Spike the sample aliquots ==> add different volume of
standards with the same concentration to the aliquots of
sample
• Dilute each solution (sample + standard) to a fixed volume
• Measure the analyte concentration
Standard Addition Methods
Instrumental measurements are made on each solutions to
get instrument response (S). If the instrument response is
proportional to concentration, we may write,
S = (kV
s
C
s
)/V
t
+ (kV
x
C
x
)/V
t
Where, V
x
=Volume of sample = 25 mL (suppose)
V
s
= Volume of standard = variable (5, 10, 15, 20 mL)
V
t
= Total volume of the flask = 50 mL
C
s
= Concentration of standard
C
x
= concentration of analyte in aliquot
k = proportionality constant
A plot of S as a function of V
s
is a straight line of the form,
S = mV
s
+b
Where, slope, m = (kC
s
)/V
t
and intercept, b = (kV
x
C
x
)/V
t
Now, b/m = (kV
x
C
x
)/V
t
x V
t
/(kC
s
)
C
x
= bC
s
/mV
x
Standard Addition Method
Another approach to determine C
x
• Extrapolate line on plot to xintercept
• Recall: At V
s
= 0 instrument response (relating
to concentration of x in sample)
• At xintercept, you know the volume of analyte
added to (i.e., inherent in) the sample.
• Another way: This value S = 0 (no instrument
response) no analyte present in sample
In any case, Since S = 0,
Therefore, S = (kV
s
C
s
)/V
t
+ (kV
x
C
x
)/V
t
= 0
Solve for C
x
,
C
x
=  (V
s
)
o
C
s
/ V
x
Standard Addition Methods
• In the interest of saving time or sample, it is possible to
perform standard addition analysis by using only two
increments of sample. A single addition of V
s
mL of
standard would be added to one of the two samples
and we can write, S
1
= (kV
x
C
x
)/V
t
and S
2
= (kV
x
C
x
)/V
t
+
(kV
s
C
s
)/V
t
S
S
k V C V C
V
X
V
kV C
V C
V C
V C
V C
S S
S
C
S VC
V S S
x x s s
t
t
x x
s s
x x
s s
x x
x
s
x
2
1
2 1
1
1
2 1
1
=
+
= +
=
÷
=
÷
( )
( )
Internal standard Method
An Internal Standard is a substance that is added
in a constant amount to all samples, blanks and
calibration standards in an analysis.
Calibration involves plotting the ratio of the
analyte signal to the internal standard signal as a
function of analyte concentration of the standards.
This ratio for the samples is then used to obtain
their analyte concentrations from a calibration
curve.
Internal standard can compensate for several
types of both random and systematic errors.
Sampling and
Method of Least Squares
Sampling is one of the most important operations in a chemical analysis.
Chemical analyses use only a small fraction of the available sample. The
fractions of the samples that collected for analyses must be representative
of the bulk materials.
Knowing how much sample to collect and how to further subdivide the
collected sample to obtain a laboratory sample is vital in the analytical
process.
All three steps of sampling, standardization, and calibration require a
knowledge of statistics.
Analytical samples and methods
Sample size Type of analysis
> 0.1g Macro
0.01~0.1g Semimicro
0.0001~0.01g Micro
< 10
–4
g Ultramicro
Analytical level Type of constituent
1%~100% Major
0.01%(100ppm)~1% Minor
1ppb~100ppm Trace
<1 ppb Ultratrace
Interlaboratory error as a function of analyte concentration.
Note that the relative standard deviation dramatically
increase as the analyte concentration decreases.
W. Horowitz, Anal. Chem., 1982, 54, 67A76A.
Real Samples
Matrix is the medium containing analyte.
A matrix effect is a change in the analytical
signal caused by anything in the sample other
than analyte.
Sample are analyzed, but constituents or
concentrations are determined.
Sampling is the process by which a sample population is
reduced in size to an amount of homogeneous material that
can be conveniently handled in the lab and whose
composition is representative of the population (unbiased
estimate of population mean).
Ex.
Population : 100 coins
Each coin is a sampling unit or an
increment
Gross sample: 5 coins
the collection of individual sampling units
or increments
Lab sample : the gross sample is reduced in size and
made homogeneous
Identify the population
Collect a gross sample
Reduce the gross
sample to a lab sample
Steps in obtaining a lab sample ( a few grams ~ a few hundred grams).
Lab sample may constitute as little as 1 part in 10
7
or 10
8
of the bulk material.
QUARTERING SAMPLES
A method of obtaining a representative sample for analysis or test of an
aggregate with occasional shovelsful, of which a heap or cone is formed,
This is flattened out and two opposite quarter parts are rejected. Another
cone is formed from the remainder which is again quartered, the process
being repeated until a sample of the required size is left.
The procedures vary somewhat, depending upon the size of the sample.
Quartering Method
Mix samples thoroughly. Pour it onto a
large flat surface.
Divide the sample into four equal parts.
Save the 2 opposite quarters. Be sure to
save the fine material at the bottom of the
saved quarter. If the sample is still to large,
divide the sample again.
Save
Save
Discard
Discard
Sampling
Sampling is the process of extracting from a large quantity of material a small portion,
which is truly representative of the composition of the whole material.
1) Three main group of sampling:
1> Census : all the material is examined ¬ impracticable
2> Casual sampling on an ad hoc basis ¬ unscientific
3> Statistical sampling
2) Sampling procedure
Bulk sample
homogeneous or heterogeneous
Increment
Gross sample
Sub sample
Analysis sample
Census vs Random sampling
Census
A complete enumeration, usually of a population, but also businesses and commercial
establishments, farms, governments, and so forth.
A complete study of the population as compared to a sample.
Random sampling
A commonly used sampling technique in which sample units are selected so that all
combinations of n units under consideration have an equal chance of being selected as
the sample.
(Statistical sampling meaning) A sampling method in which every possible sample has
the same chance of being selected.
3) Sampling statistics :
Total error = sampling error + analytical error
s
T
= ( s
S
2
+ s
A
2
)
1/2
In designing a sampling plan the following points should be
considered.
1> the number of samples to be taken
2> the size of the sample
3> should individual samples be analysed
or should a sample composed of two or more increments (composite)
be prepared.
How much should be analyzed ?
mR
2
~ K
S
where m = mass of each sample analyzed, R= desired relative SD.
How many portions should be analyzed ?
e = (ts
S
) / (n)
1/2
n = t
2
s
S
2
/ e
2
where n = the number of samples needed
t = Student’s t for the 95% confidence level and n÷1
degree of freedom.
Statistics of sampling segregated materials
s
S
2
= [A / mn] + [ B / n]
where s
S
is the standard deviation of n samples, each of mass m.
The constant A and B are properties of the bulk material and must be
measured in preliminary experiments.
Sampling uncertainties
Both systematic and random errors in analytical data can be traced to
instrument, method, and personal causes. Most systematic errors can be
eliminated by exercising care, by calibration, and by the proper use of
standards, blanks, and reference materials.
For random and independent uncertainties, the overall standard deviation
s
0
for an analytical measurement is related to the standard deviation of
the sampling process s
s
and the the standard deviation of the method s
m
by the relationship
s
0
2
= s
s
2
+ s
m
2
When s
m
< s
s
/3, there is no point in trying to improve the measurement
precision.
Size of the gross sample
Basically, gross sample weight is determined by
(1) The uncertainty that can be tolerated between the
composition of the gross sample and that of the whole,
(2) The degree of heterogeneity of the whole,
(3) The level of particle size at which heterogeneity begins.
To obtain a truly representative gross sample, a certain
number N of particles must be taken. The number of
particles required in a gross sample ranges from a few
particles to 10
12
particles.
Sampling homogeneous solutions of liquids and
gases
Well mixed solutions of liquids and gases require only a
very small sample because they are homogeneous
down to the molecular level.
Homogeneous ?
Which portion?
Flowing stream?
Gas sampling : Sampling bag (Tedlar® bag)
with a Teflon fitting
Trap in a liquid
Adsorbed onto the surface of a solid
SPME
Tedlar® bag
http://www.tedlarbag.com/tedlarbag.asp#tedlar
Depth profile of nitrate in sediment
from freshwater Lake Sobygard in
Denmark.
Lake stratification from
spring to summer
15m
18m
Epilinion 17
o
C
Thermocline 5~17
o
C
Hypolimnion
5~6
o
C
SAMPLE PREPARATION
(1) Is the sample a Solid or a Liquid?
Liquids
(2) Are you interested in all sample components or only one or a few?
If only a few then separation is necessary by extraction or
chromatography.
(3) Is the concentration of the analytes appropriate for the
measurement technique?
If not, dilute or concentrate with extraction, evaporation, lyophilization.
(4) Is sample unstable ?
If yes, derivatize, cool, freeze, store in dark
(5) Is the liquid or solvent compatible with the analytical method?
If not, do solvent exchange with extraction, distillation, lyophilization.
http://www.trincoll.edu/~dhenders/textfi~1/Chem%20208%20notes/sample_preparation.htm
Sampling particulate solids
Identify the population to be analyzed
Randomly collect N particles to give a gross sample
Reduce particle size of gross sample and homogenize
Randomly collect N particles
Is this sample of a suitable
size for the lab
Store the lab sample
Remove portions of the lab sample for analysis
No
Steps in sampling and measurement of salt in a potato chip. Step 1 introduces the
sampling variance. Steps 2 to 4 introduce the analytical variance.
Linear regression
Linear regression uses the method of least squares to
determine the best linear equation to describe a set of x and
y data points. The method of least squares minimizes the
sum of the square of the residuals  the difference between a
measured data point and the hypothetical point on a line. The
residuals must be squared so that positive and negative
values do not cancel. Spreadsheets will often have builtin
regression functions to find the best line for a set of data.
A common application of linear regression in analytical
chemistry is to determine the best linear equation for
calibration data to generate a calibration or working curve.
The concentration of an analyte in a sample can then be
determined by comparing a measurement of the unknown to
the calibration curve.
Calibration curve for the
determination of isooctane in a
hydrocarbon mixture.
The slopeintercept form of a
straight line.
Leastsquare curve fitting.
Linear leastsquares analysis
gives you the equation for the
best straight line among a set of
x, y data points when the x data
contain negligible uncertainty.
Determining the best line for calibration data is done using linear
regression.
Equation of calibration line:
y(±s
y
) = [m(±s
m
)]x + [b(±s
b
)]
The Least squares method finds the sum of the squares of the
residuals ss
resid
and minimizes these according to the minimization
technique of calculus.
ss
resid
= E[y
i
÷(b+mx
i
)]
2
A total sum of the squares :
ss
tot
= s
yy
= E(y
i
÷y)
2
= Ey
i
2
÷ Ey
i
2
/N
The coefficient of determination (R
2
) is a measure of the fraction of the
total variation in y that can be explained by the linear relationship
between x and y.
R
2
= 1 ÷ (ss
resid
/ ss
tot
)
The closer R
2
is to unity, the better the linear model explains the y
variations.
Correlation coefficient
Pearson correlation coefficient :
r = E[(x
i
÷x)(y
i
÷y) / ns
x
s
y
]
= [Ex
i
y
i
÷(nxy)] / [(Ex
i
2
÷nx
2
)(Ey
i
2
÷ny
2
)]
1/2
= (nEx
i
y
i
÷Ex
i
Ey
i
) / [{nEx
i
2
÷(Ex
i
)
2
}{nEy
i
2
÷(Ey
i
)
2
}]
1/2
r
max
= 1, r
min
= ÷1, r = 0 (when xy=0),
0.90 < r < 0.95 linearity
0.95 < r < 0.99 good linearity
0.99 < r excellent linearity
For the linear equation: y = mx + b Useful quantities:
Where:
N is the number of calibration data points.
L is the number of replicate measurements of the unknown.
and is the mean of the unknown measurements.
Standard deviation of the residuals:
Standard deviation of the intercept:
Standard deviation of the slope:
Standard deviation of a unknown read from
a calibration curve:
Chapter 2
Chemical Apparatus, and Unit Operation of
Analytical Chemistry
Classifying Chemicals
1. Reagent Grade: Reagent grade chemical conform to the minimum
standards set forth by the Reagent Chemical committee of the American
Chemical Society and are used wherever possible in analytical work.
2. Primary Standard Grade: Extraordinary purity is required for a primary
standard. Primary standard reagent is carefully analyzed and the assay is
printed on the container label.
3. SpecialPurpose Reagent: chemicals that have been prepared for a
specific application. Included among these are solvents for
spectrophotometry and highperformance liquid chromatography.
3. SpecialPurpose Reagent: chemicals that have been prepared for a
specific application. Included among these are solvents for
spectrophotometry and highperformance liquid chromatography.
2. Primary standard Grade: Extraordinary purity is required for a primary
standard. Primary standard reagent is carefully analyzed and the assay is
printed on the container label.
Rules for Handling Reagents and
Solutions
1. Select the best grade of chemical available for analytical
work.
2. Replace the top of every container immediately after
removal of the reagent.
3. Hold the stoppers of reagent bottles between your
fingers.
4. Never return any excess reagent to a bottle.
5. Never insert spatulas, spoons, or knives into a bottle
that contains a solid chemicals.
6. Keep the reagent shelf and the laboratory balance clean
and neat.
7. Observe regulations concerning the disposal of surplus
reagents and solutions.
Cleaning and Marking Laboratory Ware
Every beaker, flask, or crucible that will contain the sample
must be thoroughly cleaned before being used. The apparatus
should be washed with a hot detergent solution and then
rinsed, initially with tap water and finally with several small
portions of deionized water. Organic solvents such as
benzene or acetone may be used to remove grease films.
A chemical analysis is ordinarily performed in duplicate or
triplicate. Each vessel that holds a sample must be marked so
that its content can be positively identified. Flask, beaker and
some crucibles have small etched areas on which semi
permanent mark can be made with a pencil.
Types of Analytical Balances
An analytical balance is a weighing instrument with a
maximum capacity that ranges from 1 g to a few
kilograms with a precision of at least 1 part in 10
5
at
maximum capacity.
Macrobalances have a maximum capacity ranging
between 160 and 200 g; measurement can be made
with a standard deviation of ±0.1mg.
Semimicroanalytical balances have a maximum
load of 10 to 30 g with a precision of ±0.01mg.
Microanalytical balance has a capacity of 1 to 3 g
and a precision of ±0.001mg.
Desiccators and Desiccants
Oven drying is the most common way of removing
moisture from solids. This approach is not
appropriate for substances that decompose or for
those from which water is not removed at the
temperature of the oven.
Dried material are stored in desiccator while they cool
so as to minimize the uptake of moisture. The base
section of the desiccator contains a chemical drying
agent (desiccants) such as anhydrous calcium
chloride, calcium sulfate, magnesium perchlorate or
phosphorus pentoxide.
Desiccator
Weighing by Difference
Weighing by difference is a simple method for
determining a series of sample weights. First
the bottle and its contents are weighed. One
sample is then transferred from the bottle to a
container; gentle tapping of the bottle with its
top and slight rotation of the bottle control over
the amount of sample removed. Following
transfer, and its residual contents are
weighed. The mass of the sample is the
difference between the two weighings.
Weighing bottles
Simple Crucibles
Simple crucibles serve only as containers. Porcelain, aluminum
oxide, silica and platinum crucibles maintain constant mass and
are used principally to convert a precipitate into a suitable
weighing form. The solid is first collected on filter paper. The
filter and contents are then transferred to a weighed crucible,
and the paper is ignited.
Filtering Crucibles
Filtering crucibles serve not only as containers but also as
filters. A vacuum is used to hasten the filtration, a tight seal
between crucible and filtering flask is accomplished with any of
the several types of rubber adapters.
Filtering Crucible
Sinteredglass Crucibles
Sinteredglass crucibles are manufactured in fine, medium, and
coarse porosities. The upper temperature limit for sintered
glass crucible is ordinarily about 200
o
C. Filtering crucibles
made entirely of quartz can tolerate substantially higher
temperatures.
Filter Paper
Paper is an important filtering medium. Ashless paper is
manufactured from cellulose fibers that have been treated with
hydrochloric and hydrofluoric acids to remove metallic
impurities and silica, ammonia is then used to neutralize the
acids. It is necessary to destroy the paper by ignition if the
precipitate collected on it is to be weighed.
Decantation and transferring precipitate
Folding and seating a filter paper
Vacuum Filtration
Heating Equipment
Many precipitate can be weighed directly after being brought to
constant mass in a low temperature drying oven. Such an oven
is electrically heated and capable of maintaining a constant
temperature to within 1
o
C. The maximum attainable
temperature ranges from 140 to 260
o
C, depending on make
and model, for many precipitate 110
o
C is a satisfactory drying
temperature.
Microwave laboratory ovens are currently appearing on the
market. Where applicable, these greatly shorten drying cycles.
Muffle furnace (a heavy duty electric furnace) is capable of
maintaining controlled temperatures of 1100
o
C or higher. Long
handled tongs and heat resistance gloves are needed for
protection.
Volume Measurement
 Pipets
 Burets
 Volumetric flask
 Measuring cylinder
Typical pipets
Automatic pipet
Burets and volumetric flask
Reading a buret
Using pipet
Titration
Calibrating Glassware
Volumetric glassware is calibrated by
measuring the mass of a liquid (water) of
known density and temperature that is
contained in the volumetric ware.
Laboratory Notebook
1. Record all data and observations
2. Supply each entry with a heading
3. Date each page of the notebook
4. Never attempt to erase an incorrect entry,
cross it out with single horizontal line
5. Never remove a page from the notebook
6. Do not overcrowd entries
7. Keep first few pages for table of contents
Lab note book
Scales of Measurements
• (left) Carbonfiber electrode with a 100nanometerdiameter (100 × 10
−9
meter) tip extending from glass capillary.
The marker bar is 200 micrometers (200 × 10
−6
meter). [From W.H. Huang, D.W. Pang, H. Tong, Z.L. Wang,
and J.K. Cheng, Anal. Chem. 2001, 73, 1048.] (middle) Electrode positioned adjacent to a cell detects release of
the neurotransmitter, dopamine, from the cell. A nearby, larger counterelectrode is not shown. (right) Bursts of
electric current detected when dopamine is released. Insets are enlargements. [From W.Z. Wu, W.H. Huang, W.
Wang, Z.L. Wang, J.K. Cheng, T. Xu, R.Y. Zhang, Y. Chen, and J. Liu, J. Am. Chem. Soc. 2005, 127, 8914.]
The possible scale of measurements in analytical chemistry is astounding, ranging
from the atomic level to the size of galaxies!
Chapter 1: Measurements
11 SI Units
Using prefixes as multipliers
We customarily use prefixes for every third power of ten, e.g. 10
9
.
10
6
, 10
3
…
Converting between units
Example: Express the energy 20 Calories in terms of ? kilojoules (kJ)
Oops! In 1999, the $125 million Mars
Climate Orbiter spacecraft was lost
when it entered the Martian
atmosphere 100 km lower than
planned. The navigation error would
have been avoided if people had
written their units of measurement.
Engineers who built the spacecraft
calculated thrust in the English unit,
pounds of force. Jet Propulsion
Laboratory engineers thought they
were receiving the information in the
metric unit, newtons. Nobody caught
the error.
Why should we be concerned of units?
12 Chemical concentrations
A few concepts
• Solution: homogeneous mixture of two or more
substances.
• Solute: a minor species in a solution.
• Solvent: the major species in a solution.
• Concentration: how much solute is contained in
a given volume or mass of solution or solvent.
• Strong & weak electrolytes.
12 Chemical concentrations (Continued)
Molarity and Molality:
Molarity (M) is the number of moles of a
substance per liter of solution.
Molality (m) is concentration expressed as
moles of a substance per kilogram of solvent.
Problem 114. What is the formal concentration (expressed
as mol/L = M) of NaCl when 32.0 g are dissolved in water
and diluted to 0.500 L?
Answer: The molecular mass of NaCl is 58.44 g/mol
The moles of sale in 32.0 g are 32.0 g /58.44(g/mol)
= ……
1.3 Preparing solutions
con con dil dil
M V M V × = ×
• Preparing a solution with a desired molarity
• Dilution
Problem 132: A bottle of concentrated
aqueous sulfuric acid labeled 98.0wt %
H
2
SO
4
, has a concentration of 18.0 M. How
Many millilitres of reagent should be diluted to
1.000 L to give 1.00 M H
2
SO
4
?
Solution:
STATISTICAL TESTS
AND ERROR
ANALYSIS
PRECISION AND ACCURACY
PRECISION – Reproducibility of the result
ACCURACY – Nearness to the “true” value
TESTING ACCURACY
TESTING PRECISION
SYSTEMATIC / DETERMINATE ERROR
• Reproducible under the same conditions in the same
experiment
• Can be detected and corrected for
• It is always positive or always negative
To detect a systematic error:
• Use Standard Reference Materials
• Run a blank sample
• Use different analytical methods
• Participate in “round robin” experiments
(different labs and people running the same
analysis)
RANDOM / INDETERMINATE ERROR
• Uncontrolled variables in the measurement
• Can be positive or negative
• Cannot be corrected for
• Random errors are independent of each other
Random errors can be reduced by:
• Better experiments (equipment, methodology,
training of analyst)
• Large number of replicate samples
Random errors show Gaussian distribution for a
large number of replicates
Can be described using statistical parameters
For a large number of experimental replicates the
results approach an ideal smooth curve called the
GAUSSIAN or NORMAL DISTRIBUTION CURVE
Characterised by:
The mean value – x
gives the center of the
distribution
The standard
deviation – s
measures the width of
the distribution
The mean or average, x
÷ the sum of the measured values (x
i
) divided by the
number of measurements (n)
n
x
x
n
1 i
i
_
¿
=
=
The standard deviation, s
÷ measures how closely the data are clustered about
the mean (i.e. the precision of the data)
2
i
i
1 n
x x
s
÷


.

\

÷
=
÷
NOTE: The quantity “n1” = degrees of freedom
• Variance
• Relative standard deviation
• Percent RSD / coefficient of variation
x
s
RSD=
Other ways of expressing the precision of the data:
Variance = s
2
100
x
s
%RSD × =
POPULATION DATA
For an infinite set of data,
n → ∞ : x → µ and s → σ
population mean population std. dev.
The experiment that produces a small
standard deviation is more precise .
Remember, greater precision does not
imply greater accuracy.
Experimental results are commonly
expressed in the form:
mean ± standard deviation
s x
_
±
The more times you measure, the more confident you
are that your average value is approaching the “true”
value.
The uncertainty decreases in proportion to
n 1/
EXAMPLE
Replicate results were obtained for the analysis of
lead in blood. Calculate the mean and the standard
deviation of this set of data.
Replicate [Pb] / ppb
1 752
2 756
3 752
4 751
5 760
Replicate [Pb] / ppb
1 752
2 756
3 752
4 751
5 760
n
x
x
i
_
¿
=
( )
2
i
1 n
x x
s
÷
÷
=
¿
NB DON‟T round a
std dev. calc until
the very end.
Also:
x
s
RSD=
100
x
s
%RSD × =
0.00500
754
3.77
= =
0.500% 100
754
3.77
= × =
Variance = s
2
( ) 14.2 3.77
2
= =
754 x =
3.77 s =
754 ± 4 ppb Pb
The first decimal place
of the standard
deviation is the last
significant figure of the
average or mean.
Lead is readily absorbed through the gastro intestinal tract. In blood,
95% of the lead is in the red blood cells and 5% in the plasma. About
7090% of the lead assimilated goes into the bones, then liver and
kidneys. Lead readily replaces calcium in bones.
The symptoms of lead poisoning depend upon many factors, including
the magnitude and duration of lead exposure (dose), chemical form
(organic is more toxic than inorganic), the age of the individual
(children and the unborn are more susceptible) and the overall state of
health (Ca, Fe or Zn deficiency enhances the uptake of lead).
Pb – where from?
• Motor vehicle emissions
• Lead plumbing
• Pewter
• Leadbased paints
• Weathering of Pb minerals
European Community Environmental
Quality Directive – 50 µg/L in drinking water
World Health Organisation – recommended
tolerable intake of Pb per day for an adult –
430 µg
Food stuffs < 2 mg/kg Pb
Next to highways 20950 mg/kg Pb
Near battery works 34600 mg/kg Pb
Metal processing sites 452714 mg/kg Pb
CONFIDENCE INTERVALS
n
ts
x μ
_
± =
The confidence interval is given by:
where t is the value of student‟s t taken from the table.
The confidence interval is the expression stating that
the true mean, µ, is likely to lie within a certain
distance from the measured mean, x.
– Student‟s t test
A „t‟ test is used to compare sets of measurements.
Usually 95% probability is good enough.
Example:
The mercury content in fish samples were determined
as follows: 1.80, 1.58, 1.64, 1.49 ppm Hg. Calculate the
50% and 90% confidence intervals for the mercury
content.
n
ts
x
_
± = μ
50% confidence:
t = 0.765 for n1 = 3
( )( )
4
0.131 0.765
1.63 ± = μ
05 . 0 1.63 ± = μ
There is a 50% chance that the true
mean lies between 1.58 and 1.68
ppm
Find x = 1.63
s = 0.131
n
ts
x
_
± = μ
90% confidence:
t = 2.353 for n1 = 3
( )( )
4
0.131 2.353
1.63 ± = μ
15 . 0 1.63 ± = μ
There is a 90% chance that the true
mean lies between 1.48 and 1.78 ppm
x = 1.63
s = 0.131
1.63
1.68
1.48
1.58
1.78
90%
50%
Confidence intervals  experimental uncertainty
1) COMPARISON OF MEANS
n
s
x value known
t
calc
÷
=
Statistical tests are giving only probabilities.
They do not relieve us of the responsibility of interpreting
our results!
Comparison of a measured result with a „known‟
(standard) value
t
calc
> t
table
at 95% confidence level
¬ results are considered to be different
¬ the difference is significant!
APPLYING STUDENT‟S T:
For 2 sets of data with number of measurements n
1
, n
2
and means
x
1
, x
2
:
Where S
pooled
= pooled std dev. from both sets of data
2 n n
1) (n s 1) (n s
s
2 1
2
2
2 1
2
1
pooled
÷ +
÷ + ÷
=
2 1
2 1
pooled
2 1
calc
n n
n n
s
x x
t
+
÷
=
2) COMPARISON OF REPLICATE MEASUREMENTS
t
calc
> t
table
at 95% confidence level
¬ difference between results is significant.
Degrees of freedom
= (n
1
+ n
2
– 2)
Compare two sets of data when one sample has been
measured many times in each data set.
3) COMPARISON OF INDIVIDUAL DIFFERENCES
e.g. use two different analytical methods, A and B, to make single
measurements on several different samples.
n
s
d
t
d
calc
=
t
calc
> t
table
at 95% confidence level
¬ difference between results is significant.
1 n
) d (d
s
2
i
d
÷
÷
=
¿
Where
d = the average difference between
methods A and B
n = number of pairs of data
Perform t test on individual differences between results:
Compare two sets of data when many samples have
been measure only once in each data set.
Example:
(d
i
)
Are the two methods used comparable?
1 n
) d (d
s
2
i
d
÷
÷
=
¿
( ) ( ) ( ) ( ) ( ) ( )
1 6
04 . 0 02 . 0 11 . 0 11 . 0 22 . 0 02 . 0
s
2 2 2 2 2 2
d
÷
+ + + + ÷ + ÷
=
12 . 0 s
d
=
n
s
d
t
d
calc
=
( )
( )
6
0.12
0.06
t
calc
=
2 . 1 t
calc
=
t
table
= 2.571 for 95%
confidence
t
calc
< t
table
¬ difference between results
is NOT significant.
2
2
2
1
calc
s
s
F =
F
calc
> F
table
at 95% confidence level
¬ the std dev.‟s are considered to be different
¬ the difference is significant.
F TEST
COMPARISON OF TWO STANDARD
DEVIATIONS
Q TEST FOR BAD DATA
range
gap
Q
calc
=
The range is the total spread
of the data.
The gap is the difference
between the “bad” point and
the nearest value.
Example:
12.2 12.4 12.5 12.6 12.9
Gap
Range
If Q
calc
> Q
table
÷ discarded
questionable point
EXAMPLE:
The following replicate analyses were obtained when
standardising a solution: 0.1067M, 0.1071M, 0.1066M and 0.1050M.
One value appears suspect. Determine if it can be ascribed to
accidental error at the 90% confidence interval.
Arrange in increasing order:
Q =
Gap
Range
Analytical Figures of Merit
―Indicate a characteristic of an instrumental
technique for a given analyte‖
“7”
Accuracy, Precision, SignaltoNoise Ratio
Sensitivity, Limit of Detection
Linearity, Linear Dynamic Range
Accuracy
Indicates how close the measured value is
to the true analytical concentration
Requires a Standard Reference Material
(SRM) of other official measure
NIST: National Institute of Standards and
Technology
Accuracy
Most commonly reported as percent error
│C
m
 C
t
│
C
t
where:
C
m
= measured concentration
C
t
= true concentration
x 100%
Precision
Indicates the reproducibility of repetitive
measurements of equivalent samples
May be expressed as:
1. Standard Deviation (s or σ)
2. Relative Standard Deviation (RSD)
3. Confidence Limits
Precision
Standard Deviation
For an infinite number of measurements (σ)
For a finite number of measurements (s)
Standard Deviation
Note that both s and σ have the same units
as the original values
How many values should be obtained?
Rule of thumb: 16
0
5
10
15
20
25
30
35
40
45
96.0 97.0 98.0 99.0 100.0 101.0 102.0 103.0 104.0
Value
P
o
p
u
l
a
t
i
o
n
Total Population = 1000
0.00%
0.10%
0.20%
0.30%
0.40%
0.50%
0.60%
0.70%
0.80%
0.90%
1.00%
0 5 10 15 20 25 30 35 40 45 50
Number of Samples
E
r
r
o
r
i
n
M
e
a
n
How far is the measured mean from the true value?
0.0%
10.0%
20.0%
30.0%
40.0%
50.0%
60.0%
70.0%
0 5 10 15 20 25 30 35 40 45 50
Number of Samples
E
r
r
o
r
i
n
S
t
d
.
D
e
v
.
How far is s from σ?
Short Cut: σ ≈ 1/5 (peaktopeak noise)
Relative Standard Deviation
RSD = σ/mean
Where the mean may be the signal or the
analyte concentration. RSD is a unitless
value, so σ must have the same units as
the mean.
RSD is often reported as %RSD, and may be
used to compare different techniques.
Confidence Limits
Define an interval that encloses
the true value (C
t
) with a
specified level of confidence.
1. C
m
± σ 66.7% Confidence Level
2. C
m
± 2σ 95% Confidence Level
3. C
m
± 3σ 99.0% Confidence Level
Signal to noise Ratio (S/N)
S/N = S
m
/σ = 1/RSD
Notes:
1. N = noise (σ)
2. S/N is unitless
3. Always try top maximize S/N
4. S/N is used to compare instruments
5. A plot of S/N versus an instrumental parameter reaches
a maximum at the optimum value for that parameter
Sensitivity
Experimental slope of a calibration curve
m = ΔS/ΔC
Sensitivity is almost always specific for one
particular instrument.
0
10
20
30
40
50
60
70
80
90
100
0 2 4 6 8 10 12
Concentration (ppm)
S
i
g
n
a
l
(
V
)
m
LOD
LDR
Limit of Detection
The analyte concentration yielding an
analytical signal equal to 3 times the
standard deviation in the blank signal.
LOD = 3 x σ
bl
/ m
By definition, the LOD has just one
significant figure!!
Linearity
Measure of how well the observed data
follows a straight line.
S
A
= mC
S
A
= Analytical Signal
m = calibration sensitivity
Remember S
A
= S
tot
 S
bl
Linearity
Plot log(S) versus log(C)
log(S
A)
= log(m) + log(C)
The slope of this plot should be 1.00
A calibration curve is defined as linear if the
loglog plot has a slope in the range 0.951.05
Linear Dynamic Range
The concentration range over which the
calibration curve is linear
Lower End → LOD
Upper End
Analyte Concentration where the observed
signal falls 5% below the extrapolated line
LDR Units are
―orders of magnitude‖
or
―decades‖
of analyte concentration
LDR is easiest to observe on loglog plot
If linearity is poor, define an analytically
useful range (AUR)
Other figures of merit may be calculated,
but these 7 are sufficient.
Selectivity and Resolution may be useful
in cases where more than one analyte is
determined in the same sample.
141
Chapter 2
Data Handling
142
Accuracy and Precision
Accuracy can be defined as the degree of
agreement between a measured value and
the true or accepted value. As the two values
become closer, the measured value is said to
be more accurate.
Precision is defined as the degree of
agreement between replicate measurements
of the same quantity.
143
Assuming the correct or accepted value is
represented by the center of the circles
below, if all values occurred within, for
example, the red circles, results are
precise but not accurate. If all values
occurred within the yellow circle, results
are both accurate and precise. If results
were scattered randomly, results are
neither precise nor accurate.
144
145
Example
The weight of a person was measured five times
using a scale. The reported weights were 84, 83,
84, 85, and 84 kg. If the weight of a person is 76
kg weighed on a standard scale , then we know
that the results obtained using the first scale is
definitely not accurate.
However, the values of the weights for the five
replicate measurements are very close and
reproducible. Therefore measurements are
precise. Therefore, a measurement could be
precise but not accurate.
146
Significant Figures
At the most basic level, Analytical Chemistry
relies upon experimentation;
experimentation in turn requires
numerical measurements. And
measurements are always taken from
instruments made by other workers.
Significant figures are concerned with
measurements not exact countings.
147
Some information about
measurements
1) Examples we will study include the metric ruler,
the graduated cylinder, and the scale.
2) Because of the involvement of human beings,
NO measurement is exact; some error is always
involved. This means that every answer in
science has some uncertainty associated with it.
We might be fairly confident we have the correct
answer, but we can never be 100% certain we
have the EXACT correct answer.
148
3) Measurements always have two parts  a
numerical part (sometimes called a factor) and a
dimension (a unit). The reason for this is that we
are measuring quantities  length, elapsed time,
temperature, mass, etc. Not only do we have to
tell how much there is, but we have to tell how
much of what.
Measuring gives significance (or meaning) to each
digit in the number produced. This concept of
significance, of what is and what is not
significant is VERY IMPORTANT.
149
The concept of significant figures (or
significant digits) is important. A
measurement can be defined as the
comparison of the dimensions of an object
to some standard.
The dimensions of an object refer to some
property the object possesses. Examples
include mass, length, area, density, and
electrical charge. Dimensions are often
called units.
150
Identifying significant digits
The following rules are helpful in identifying significant
digits
1. Digits other than zero are significant.
e.g., 42.1m has 3 sig figs.
2. Zeroes are sometimes significant, and sometimes
they are not.
3. Zeroes at the beginning of a number (used just to
position the decimal point) are not significant.
e.g., 0.025m has 2 sig figs. In scientific notation, this can
be written as 2.5*10
2
m
151
4. Zeroes between nonzero digits are significant.
e.g., 40.1m has 3 sig figs
5. Zeroes at the end of a number that contains a
decimal point are significant.
e.g., 41.0m has 3 sig figs, while 441.20m has 5.
In scientific notation, these can be written
respectively as
4.10*10
1
and 4.4120*10
2
6. Zeroes at the end of a number that does not
contain a decimal point may or may not be
significant. If we wish to indicate the number of
significant figures in such numbers, it is common
to use the scientific notation.
152
e.g., The quantity 52800 km could be having 3, 4,
or 5 sig figs—the information is insufficient for
decision. If both of the zeroes are used just to
position the decimal point (i.e., the number was
measured with estimation ±100), the number is
5.28×10
4
km (3 sig figs) in scientific notation. If
only one of the zeroes is used to position the
decimal point (i.e., the number was measured
±10), the number is 5.280×10
4
km (4 sig figs). If
the number is 52800±1 km , it implies
5.2800×10
4
km (5 sig figs).
153
Exact Numbers
Exact numbers can be considered as having an unlimited
number of significant figures. This applies to defined
quantities too. e.g.,
1. The rules of significant figures do not apply to (a) the
count of 47 people in a hall, or (b) the equivalence: 1
inch = 2.54 centimeters.
2. In addition, the power of 10 used in scientific notation
is an exact number, i.e. the number 10
3
is exact, but the
number 1000 has 1 sig fig.
It actually makes a lot of sense to write numbers derived
from measurements in scientific notation, since the
notation clearly indicates the number of significant digits
in the number.
154
Look at the following example
155
The general rule for estimation of the last
digit is to record to 1/10th of the smallest
division of the measuring device. So, for
the common centimeterruler, in which the
smallest division is 1mm, the estimate for
the last digit can be to 0.1mm which is
1/10th, of the smallest division.
156
Suppose a student records a length of
26.2mm with a ruler (between the red
marks). In his/her judgment, the length is
greater than 26.1mm but less than
26.3mm, and so the best estimate is
26.2mm. The measurement can be written
as 26.2±0.1 mm. The number 26.2mm
contains three significant figures. Although
the last digit, 2, is an estimate, it is
considered to be a significant figure for the
measurement.
157
How do you read it?
How do you read it?
158
159
160
161
162
Math With Significant Figures
Addition and Subtraction
In mathematical operations involving significant figures, the
answer is reported in such a way that it reflects the
reliability of the least precise operation. Let's state that
another way: a chain is no stronger than its weakest link.
An answer is no more precise that the least precise
number used to get the answer. Let's do it one more
time: imagine a team race where you and your team
must finish together. Who dictates the speed of the
team? Of course, the slowest member of the team. Your
answer cannot be MORE precise than the least precise
measurement.
163
For addition and subtraction, look at the decimal
portion (i.e., to the right of the decimal point) of
the numbers ONLY. Here is what to do:
1) Count the number of significant figures in the
decimal portion of each number in the problem.
(The digits to the left of the decimal place are not
used to determine the number of decimal places
in the final answer.)
2) Add or subtract in the normal fashion.
3) Round the answer to the LEAST number of
places in the decimal portion of any number in
the problem.
164
Find the formula weight for Ag
2
MoO
4
given the
following atomic weights: Ag = 107.870, Mo =
95.94, O = 15.9994.
The number with the least number of digits after
the decimal point is 95.94 which has two digits
for expression of precision. Also, it is the number
with the highest uncertainty. The atomic weights
for Ag and O have 3 and 4 digits after the
decimal point. Therefore if we calculate the
formula weight we will get 375.6776. However,
the answer should be reported as 375.68 ( i.e. to
the same uncertainty of the least precise value.
165
Multiplication and Division
In mathematical operations involving significant
figures, the answer is reported in such a way
that it reflects the reliability of the least precise
operation. Let's state that another way: a chain
is no stronger than its weakest link. An answer is
no more precise that the least precise number
used to get the answer. Let's do it one more
time: imagine a team race where you and your
team must finish together. Who dictates the
speed of the team? Of course, the slowest
member of the team. Your answer cannot be
MORE precise than the least precise
measurement.
166
The following rule applies for multiplication and
division:
The number having the least number of significant
figures is called the KEY NUMBER. The LEAST
number of significant figures in any number of
the problem determines the number of
significant figures in the answer.
This means you MUST know how to recognize
significant figures in order to use this rule.
In case where two or more numbers have the
same least number of significant figures, the key
number is determined as the number of the
lowest value regardless of decimal point.
167
2.5 x 3.42.
The answer to this problem would be 8.6
(which was rounded from the calculator
reading of 8.55). Why?
2.5 is the key number which has two
significant figures while 3.42 has three.
Two significant figures is less precise than
three, so the answer has two significant
figures.
168
How many significant figures will the answer to
3.10 x 4.520 have?
You may have said two. This is too few. A
common error is for the student to look at a
number like 3.10 and think it has two significant
figures. The zero in the hundredth's place is not
recognized as significant when, in fact, it is. 3.10
is the key number which has three significant
figures.
Three is the correct answer. 14.0 has three
significant figures. Note that the zero in the
tenth's place is considered significant. All trailing
zeros in the decimal portion are considered
significant.
Example
Synthetic Chemistry Analytical Chemistry
To make a new stuff
To find out what is it
“white powder”
Qualitative
Quantitative
to identify
This white powder is
NaCl
to detect
There is a trace
admixture
of iodine in this
sample
to analyze
to determine X in a sample
to quantify
to measure
Run assay
determination
quantitation
0.375 mg/mL caffeine
Composition, %%:
Cu, 85.34(3); Zn, 11.23; Sn, 1.02(2);
Fe, 0.123(2); Mn, 0.058(5); Ni, 0.229(5); …
A chocolate story: how much caffeine?
Garbage in,
Garbage out!
Calibration
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