ANALYTICAL

CHEMISTRY
Course Tutor: Dr. Kashif Gul
Course: Special Topics in Analytical
Chemistry
Where to Sample the Target Population

Homogeneous and heterogeneous populations.

Homogenization destroys information about the analytes spatial or temporal distribution
within the target population.
Random Sampling
Selected by using chance or random numbers. Each individual subject (human or otherwise)
has an equal chance of being selected. Eliminates bias in the selection process. Also known as
probability sampling.
Number each frame unit from 1 to N. Use a random number table or a random number
generator to select n distinct numbers between 1 and N, inclusively.
Easier to perform for small populations.
Judgmental Sampling
Use of available information about the target population to help select samples.
More biased than random sampling; however, fewer samples are required.
Limits the number of independent variables influencing the results of an analysis.
 Systematic Sampling
Population elements are an ordered sequence (at least, conceptually).
The first sample element is selected randomly from the first k population elements.
Thereafter, sample elements are selected at a constant interval, k, from the ordered
sequence frame.
Convenient and relatively easy to administer.

N
k = ,
n
where :
n = sample size
N = population size
k = size of selection interval
Stratified Random Sample
Stratified sampling reduces sampling error.
A stratum is a subset of the population that share at least one common
characteristic; such as males and females.
Identify relevant stratums and their actual representation in the population.
Random sampling is then used to select a sufficient number of subjects from each
stratum.
Stratified sampling is often used when one or more of the stratums in the
population have a low incidence relative to the other stratums.

Convenience Sampling
Use subjects that are easily accessible
Sampling Distribution of Mean

Proper analysis and interpretation of a sample statistic requires knowledge of its

distribution.

Calculatex
toestimate
Population Sample
Process of x
Inferential Statistics
(parameter) (statistic)

Selecta
randomsample
Central Limit Theorem

If x is the mean of a random sample of size

n from a population with mean of and
standard deviation of , then as n increases
the distribution of x approaches a normal
distribution with mean and
x

standard deviation x .
n
Z Formula for Sample Means

X X
Z
X

X


n
 Sampling from a Finite Population

Finite Correction Factor Nn

Modified Z Formula N 1

X
Z
Nn
n N 1
Sampling Distribution of p

Sample Proportion
X
p$
n
where:
X numberofitemsinasamplethatpossessthecharacteristic
n = numberofitemsinthesample

Sampling Distribution
Approximately normal if nP > 5 and nQ > 5 (P
is the population proportion and Q = 1 - P.) P Q
The mean of the distribution is P.
The standard deviation of the distribution is n
 Analytical Samples and Methods
Compromise between the accuracy
required and the economics available for the
analysis.
The number of samples that will be
analyzed determines the amount of time that
can be spend in preliminary operations such
as assembling and calibrating instruments
and equipment and preparing standard
solutions.
Complexity of the sample and the number
of components in the sample will also
influence the choice of method.
 Types of Samples and Methods
Sample Size:

Constituent Types:
eneral problem in trace procedures is that the reliability of results usually decreases
matically with a decrease in analyte level.
 Sampling
The process by which a representative
Who = Population:
fraction is acquired from a sampling all individuals elements of
frame is termed as sampling. interest
What = Parameter
Accuracy limiting step of the procedure. Characteristic of
population
This statement is especially true when Problem: cant study/survey
the material to be analyzed is a large whole pop
Solution: Use a sample for the
and inhomogeneous liquid, such as a who
subset, selected from
lake, or an inhomogeneous solid, such as population
an ore, a soil, or a piece of animal tissue. calculate a statistic for the
what
 Types of
Statistics
Descriptive statistics:
Organize and summarize scores from
samples
Inferential statistics:
Infer information about the
population based on what we
know from sample data
Decide if an experimental
manipulation has had an effect

An element is an object on which a measurement is taken.

A population is a collection of elements about which we wish to make an inference.

Sampling units are non-overlapping collections of elements from the population

that cover the entire population.
 A sampling frame is a list of sampling units.
A sample is a collection of sampling units drawn from a sampling frame.
Parameter: numerical characteristic of a population
Statistic: numerical characteristic of a sample

The deviation between an estimate from
an ideal sample and the true
population value is the sampling
error.
Almost always, the sampling frame
does not match up perfectly with
the target population, leading to
errors of coverage.
The population of interest is usually too
large to attempt to survey all of its
members.
A carefully chosen sample can be used to
represent the population.
The sample reflects the characteristics of
the population from which it is drawn.
Obtaining a Representative Sample

Sampling of non-
homogeneous material where
concentrations of the analyte
differ.
Prepare a representative
sample by taking our sample
increments from different
regions of the bulk material
Statistically, the goals of the sampling process are:
1. To obtain a mean analyte concentration that is an unbiased estimate of the
population mean.
This goal can be realized only if all members of the population have an equal
probability of being included in the sample.
2. To obtain a variance in the measured analyte concentration that is an
unbiased estimate of the population variance so that valid confidence limits
can be found for the mean, and various hypothesis tests can be applied.
This goal can be reached only if every possible sample is equally likely to be
drawn.
Probability versus Non-probability

Probability Samples:
Each member of the population has a known non-zero probability of being
selected
Methods include random sampling, systematic sampling, and stratified
sampling.
Non-probability Samples:
Members are selected from the population in some nonrandom manner
Methods include convenience sampling, judgment sampling, quota
sampling, and snowball sampling.
 The Gross Sample
Miniature replica of the entire mass of material (Sampling frame) .
It should correspond to the bulk material i-e inchemical composition and in particle-size
distribution.

Size of the Gross Sample

Basically, gross sample size 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, and
(3) the level of particle size at which heterogeneity begins.
Levels of heterogeneity in homogeneous and heterogeneous solutions i-e a homogeneous
solution of a gas or liquid is heterogeneous only on the molecular scale, a particulate solid,
such as an ore or a soil have individual pieces of solid differ from each other in
composition.
Heterogeneity develops in particles that may have dimensions on the order of a centimeter
or more and may be several grams in mass.
For example, let us presume that a pharmaceutical
mixture contains just two types of homogeneous
particles: type A particles containing the active
ingredient and type B particles containing only an
inactive filler material.
p = probability of randomly drawing an A type
particle
1-p = probability of randomly drawing an B type
particle
N = particles of the mixture taken
pN = the most probable value for the number of A
type particles
(1-p)N = the most probable number of B type
particles Random sample should consist of 2500 particles.
For such a binary population, the Bernoulli
equation can be used to calculate the standard
deviation of the number of A particles drawn, A.
Let us assume that both of the components in the mixture The relative standard deviation of sampling, r .
contain the active ingredient (analyte), such that the type A
particles contain a higher percentage of analyte, PA and the

type B particles a lesser amount, PB and the overall average

percent of active ingredient is P.
Furthermore, the average density d of the particles differs from
the densities dA and dB of these components.
Sampling Homogeneous Solutions of Liquids
and Gases

Sampling Particulate Solids

Sampling Metals and Alloys

 Standardization and Calibration:

Calibration determines the relationship between the analytical response and the analyte
concentration by the use of chemical standards (Standardized chemicals).
Comparison with Standards
Direct Comparison
Null comparison or Isomation methods
Some analytical procedures involve comparing a property of the analyte (or the product
of a reaction with the analyte) with standards such that the property being tested
matches or nearly matches that of the standard.

With some modern instruments, a variation of this procedure is used to determine if an analyte
concentration exceeds or is less than some threshold level. Let us consider that the exact concentration
of aflatoxin is not needed but only the comparator is needed to indicate that the threshold has been
exceeded. Alternatively, the approximate concentration of the analyte can be determined by comparing
the color of the unknown solution with those of several standards.

In the comparison method, antibodies specific to the aflatoxin are coated on the base of a plastic
compartment or microtiter well in an array on a plate such as that shown in The aflatoxin behaves as
the antigen. During the analysis, an enzyme reaction causes a blue product to be formed. As the
amount of aflatoxin in the sample increases, the blue color decreases in intensity.
Titrations
One of most accurate of all analytical procedures.
Analyte reacts with a standardized reagent (the titrant) in a known stoichiometric manner.
Chemical equivalence is usually indicated by the color change of a chemical indicator or
by the change in an instrument response.
The amount of the standardized reagent needed to achieve chemical equivalence can then
be related to the amount of analyte present by means of the stoichiometry.
 External Standard Calibration
A series of standard solutions is prepared separately from the sample.
The standards are used to establish the instrument calibration function, which is obtained
from analysis of the instrument response as a function of the known analyte concentration.
The calibration function can be obtained graphically or in mathematical form.
Generally, a plot of instrument response versus known analyte concentrations is used to
produce a calibration curve, sometimes called a working curve.
It is often desirable that the calibration curve be linear in at least the range of the analyte
concentrations.
The Least-Squares Method

The indeterminate errors in

the measurement process result
in deviation of data points from
the line.
Regression analysis provides
a straight line taking into
consideration the uncertainties.
Assumptions of the Least-Squares Method:

Correlation analysis
Weighted least-squares analysis
Non-linear regression methods
Errors in External Standard Calibration

The same response will be obtained for

analyte concentration which is present in the
sample and in the standard.
Usually in a determination, the raw response
from the instrument is not used but instead,
the raw analytical response is corrected by
measuring a blank . The ideal blank is identical
to the sample but without the analyte.
Most often a real blank is either a solvent Matrix effects
blank, containing the same solvent in which the Experimental Variables
sample is dissolved, or a reagent blank,
Contamination
containing the solvent plus all the reagents
Systematic Errors
used in sample preparation.
Random Errors
 Figures of Merit for Analytical Methods
Analytical procedures are characterized by a number of figures of merit such as accuracy,
precision, sensitivity, detection limit, and dynamic range.
Sensitivity and Detection Limit
The calibration sensitivity is the change in the response signal per unit change in analyte
concentration.
The calibration sensitivity is thus the slope of the calibration curve i-e linear curve means
sensitivity is constant and independent of concentration whereas nonlinear, sensitivity
changes with concentration and is not a single value.
Analytical sensitivity- Analytical sensitivity is usually a strong function of concentration.
The detection limit, DL, is the smallest concentration that can be reported with a certain
level of confidence i-e the analyte concentration that produces a response equal to k times
the standard deviation of the blank sb. For methods that require a calibration curve, the
detection limit is defined in a practical sense;

where k is called the confidence factor and m is the calibration sensitivity. The factor k is
usually chosen to be 2 or 3. A k value of 2 corresponds to a confidence level of 92.1%, while
a k value of 3 corresponds to a 98.3% confidence level.

Linear dynamic range with upper and lower limit

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