Analytical Chemistry

Dr Swagat Mohapatra
UGC – Asst Professor
ICT – IOC Bhubaneswar
Analytical Chemistry
• Introduction
• Basic Tool
• Evaluation of analytical data
• Collecting & preparing samples
• Gravimetric & titrimetric methods
• Spectroscopic methods
• Chromatographic methods
• Quality Assurance
Fresenius’ analytical scheme for the gravimetric analysis of Ni in ores.

Note that the mass of nickel is not determined directly. Instead, Co and Ni are
isolated and weighed (mass A), and then Co is isolated and weighed (mass B). The
timeline shows that after digesting a sample, it takes approximately 44 hours to
complete an analysis. This scheme is an example of a gravimetric analysis
in which mass is the important measurement.
Gravimetric analysis for Ni in ores by precipitating Ni(dmg) .

• The development, in 1905, of dimethylglyoxime (dmg), a

reagent that selectively precipitates Ni2+ and Pd2+ and led to
an improved analytical method for the quantitative analysis
of nickel.
• The resulting analysis, as shown in Figure , requires fewer
manipulations and less time after completing the sample’s
dissolution.
• By the 1970s, flame atomic absorption spectrometry
replaced gravimetry as the standard method for analyzing
nickel in ores, resulting in an even more rapid analysis.
• Today, the standard analytical method utilizes an inductively
coupled plasma optical emission spectrometer.

The timeline shows that it takes approximately 4 hours

to complete an analysis after digesting the sample, which is 10x
shorter than the earlier method .
Analytical Chemistry
• Analytical chemistry is often described as the area of chemistry responsible for characterizing the composition
of matter, both qualitatively (Is there any lead in this sample?) and quantitatively (How much lead is in this
sample?).

• Typical problems on which analytical chemists work include:

1- qualitative analyses (What is present?),
2- quantitative analyses (How much is present?),
3- characterization analyses (What are the sample’s chemical and physical properties?), and
4- fundamental analyses (How does this method work and how can it be improved?).
Seven Stages of analytical method
• 1. Conception of analytical method (birth).
• 2. Successful demonstration that the analytical method works.
• 3. Establishment of the analytical method’s capabilities.
• 4. Widespread acceptance of the analytical method.
• 5. Continued development of the analytical method leads to significant improvements.
• 6. New cycle through steps 3–5.
• 7. Analytical method can no longer compete with newer analytical methods (deat
 Defination – Analytical Chemistry
• A more appropriate description of analytical chemistry is “the science of inventing and applying the concepts,
principles, and strategies for measuring the characteristics of chemical systems.”

• Analytical chemists typically operate at the extreme edges of analysis, extending and improving the ability of all
chemists to make meaningful measurements on smaller samples, on more complex samples, on shorter time
scales, and on species present at lower concentrations.

• Throughout its history, analytical chemistry has provided many of the tools and methods necessary for research
in the other traditional areas of chemistry, as well as fostering multidisciplinary research in, to name a few,
medicinal chemistry, clinical chemistry, toxicology, forensic chemistry, materials science, geochemistry, and
environmental chemistry.
Sampling
• A proper sampling strategy ensures that our samples are
representative of the material from which they are taken.
• Biased or nonrepresentative sampling, and contaminating samples
during or after their collection are sampling errors that can lead to a
significant error in accuracy. It is important to realize that sampling
errors are independent of errors in the analytical method.
The Importance of Sampling
When a manufacturer lists a chemical as ACS Reagent Grade, they must
demonstrate that it conforms to specifications set by the American Chemical
Society (ACS).

For example, the ACS specifications for NaBr require that the concentration
of iron be ≤5 ppm. To verify that a production lot meets this standard, the
manufacturer collects and analyzes several samples, reporting the average
result on the product’s label.

If the individual samples do not accurately represent the population from which they are drawn—what we call the target
population—then even a careful analysis must yield an inaccurate result. Extrapolating this result from a sample to its
target population introduces a determinate sampling error. To minimize this determinate sampling error, we must collect
the right sample.
Chemical Grades:
Most laboratory chemicals are labelled with a grade which generally indicates the purity of the chemical. There are many
different standards used in grading, however, the three you are most likely to encounter are:

(1) Reagent Grade (purest), (2) Laboratory Grade (middle tier quality), and (3) Technical Grade (least pure).

Reagent Grade
• Alternate Names: Analytical Reagent (AR), Guaranteed Reagent (GR), UNIVAR, AnalaR, Premium Reagent (PR)
• Purity Level: Very high purity, meets or exceeds standards set by American Chemical Society (ACS).

Laboratory Grade
• Alternate Names: Laboratory Reagent (LR), UNILAB, Chemically Pure (CP)
• Purity Level: Relatively high purity, exact levels of impurity unknown.

Technical Grade
• Alternate Names: Tech Grade (TG), Commercial Grade
• Purity Level: Contains impurities. Basic product specs usually available.

Other Grades
• HPLC grades; Ultra Pure reagents; AAS/ICP Standards; Ultrapure reagents; Molecular Biology grade; Reference
Standards for USP, EP and BP testing; Food Pharmaceutical grades (BP, EP, FG).
Designing a Sampling Plan
1. Where to Sample the Target Population
• Random Sampling
• Judgmental Sampling
• Systematic Sampling
• Systematic–Judgmental Sampling
• Stratified Sampling
• Convenience Sampling
2. What Type of Sample to Collect
3. How Much Sample to Collect
4. How Many Samples to Collect
5. Minimizing the Overall Variance
Where to Sample the Target Population
A sampling error occurs whenever a sample’s composition is not identical to its target population. If the
target population is homogeneous, then we can collect individual samples without giving consideration
to where to sample. Unfortunately, in most situations the target population is heterogeneous.

Random Sampling
The ideal sampling plan provides an unbiased estimate of the target population’s properties. A random
sampling is the easiest way to satisfy this requirement. Despite its apparent simplicity, a truly random
sample is difficult to collect.

Judgmental Sampling
• The opposite of random sampling is selective, or judgmental sampling in which we use prior
information about the target population to help guide our selection of samples. Judgmental sampling
is more biased than random sampling, but requires fewer samples. Judgmental sampling is useful if
we wish to limit the number of independent variables influencing our results. For example, if we are
studying the bioaccumulation of PCB’s in fish, we may choose to exclude fish that are too small or
that appear diseased.

Systematic Sampling
• Random sampling and judgmental sampling represent extremes in bias and in the number of samples
needed to characterize the target population. Systematic sampling falls in between these extremes.
In systematic sampling we sample the target population at regular intervals in space or time.
Implementing the Sampling Plan
• Solutions
1. Sample Collection
2. Sample Preservation and Preparation
• Gases
1. Sample Collection
2. Sample Preservation and Preparation
• Solids
1. Sample Collection
2. Sample Preservation
3. Sample Preparation
4. Reducing Particle Size
5. Bringing Solid Samples Into Solution
Implementing the Sampling Plan
Implementing a sampling plan normally involves three steps:
• physically removing the sample from its target population - we analyze a sample
after removing it from its target population. Because sampling exposes the target
population to potential contamination, the sampling device must be inert and clean
• preserving the sample - After removing a sample from its target population, there is
a danger that it will undergo a chemical or physical change before we can complete
its analysis. This is a serious problem because the sample’s properties no longer are
representative of the target population. To prevent this problem, we often preserve
samples before transporting them to the laboratory for analysis.
• preparing the sample for analysis - The initial sample is called the primary or gross
sample, and may be a single increment drawn from the target population, or a
composite of several increments. In many cases we cannot analyze the gross sample
without first reducing the sample’s particle size, converting the sample into a more
readily analyzable form, or improving its homogeneity.
Solutions
• Typical examples of solution samples include:
1- those drawn from containers of commercial solvents;
2- beverages, such as milk or fruit juice;
3- natural waters, including lakes, streams, seawater and rain;
4- bodily fluids, such as blood and urine;
5- suspensions; such as those found in many oral medications..

Solutions-Sample Collection
• The chemical composition of a surface water—such as a stream, river, lake, or ocean—is influenced by flow rate
and depth. Rapidly flowing shallow streams and rivers, and shallow (<5 m) lakes are usually well mixed, and show
little stratification with depth.

• To collect a grab sample, submerge a capped bottle below the surface, remove the cap and allow the bottle to fill
completely, and replace the cap. Collecting a sample this way avoids the air–water interface, which may be
enriched with heavy metals or contaminated with oil.
Sample Preservation and Preparation
To prevent a change in composition, samples are preserved by
• controlling the solution’s pH and temperature,
• by limiting its exposure to light or to the atmosphere, or
• by adding a chemical preservative.

After preserving a sample, it may be safely stored for later analysis.

Most solution samples do not need additional preparation before analysis. This is the case for samples of natural waters
and wastewaters. Solution samples with particularly complex matricies—blood and milk are two examples—may need
additional processing to separate the analytes from interferents.
Sample Preservation and Preparation
Preservation Methods and Maximum Holding Times for Selected Analytes in Natural Waters and Wastewaters

Analyte Preservation Method Maximum Holding Time

ammonia cool to 4oC; add H2SO4 to pH<2 28 days

chloride none required 28 days
metals—Cr(VI) cool to 4oC 24 hours

metals—Hg HNO3 to pH<2 28 days

metals—all others HNO3 to pH<2 6 months

nitrate none required 48 hours

1 mL of 10 mg/mL HgCl2 or immediate extraction 7 days without extraction

organochlorine pesticides
with a suitable non-aqueous solvent 40 days with extraction

pH none required analyze immediately

Gases
Typical examples of gaseous samples include

• automobile exhaust,
• emissions from industrial smokestacks,
• atmospheric gases,
• and compressed gases.

Also included in this category are aerosol particulates—the fine solid particles and liquid droplets
that form smoke and smog.
Sample Collection
• One approach for collecting a sample of urban air is to fill a stainless steel canister or a Tedlar/Teflon bag. A
pump pulls the air into the container and, after purging, the container is sealed. This method has the
advantage of simplicity and of collecting a representative sample.

• Disadvantages include the tendency for some analytes to adsorb to the container’s walls, the presence of
analytes at concentrations too low to detect with accuracy and precision, and the presence of reactive
analytes, such as ozone and nitrogen oxides, that may react with the container or that may otherwise alter
the sample’s chemical composition during storage. When using a stainless steel canister, cryogenic cooling,
which changes the sample from a gaseous state to a liquid state, may limit some of these disadvantages.
Solids
Typical examples of solid samples include
• large particulates, such as those found in ores;
• smaller particulates, such as soils and sediments;
• tablets, pellets, and capsules used for dispensing pharmaceutical products and animal feeds;
• sheet materials, such as polymers and rolled metals;
• and tissue samples from biological specimens.

Solids are usually heterogeneous and samples must be collected carefully if they are to be representative of
the target population. Let’s use the sampling of sediments, soils, and ores as a case study in how to sample
solids
Sample Collection
• Sediments from the bottom of streams, rivers, lakes, estuaries, and oceans are collected with a bottom grab
sampler, or with a corer.
• Its principal advantages are ease of use and the ability to collect a large sample.
• Disadvantages include the tendency to lose finer grain sediment particles as water flows out of the sampler, and the
loss of spatial information—both laterally and with depth—due to mixing of the sample.
UNIT -II

• Theory of Errors
• Uncertainties in measurements
• Sources & Classification of Errors
 Close-up of a burette showing the difficulty in estimating volume.
With scale divisions every 0.1 mL it is difficult to read the actual volume to better than ±0.01–0.03 mL.
Evaluating Analytical Data
When using an analytical method we make three separate evaluations of experimental error.
• First, before beginning an analysis we evaluate potential sources of errors to ensure that they will not adversely effect
our results.
• Second, during the analysis we monitor our measurements to ensure that errors remain acceptable.
• Finally, at the end of the analysis we evaluate the quality of the measurements and results, comparing them to our
original design criteria.
Characterizing Measurements and Results
One way to characterize data from multiple measurements/runs is to assume that the measurements are randomly
scattered around a central value that provides the best estimate of expected, or “true” value.

1. Measures of Central Tendency

• Mean
• Median

2. Measures of Spread
• Range
• Standard Deviation
• Variance
 Masses of Seven Circulating U. S. COINs
Coin Mass (g)
1 3.080
2 3.094
3 3.107
4 3.056
5 3.112
6 3.174
7 3.198

1. Measures of Central Tendency

• Mean ????
• Median ?????

2. Measures of Spread
• Range ????
• Standard Deviation ?????
• Variance ?????
Measures of Central Tendency
One way to characterize the data in Table is to assume that the masses are randomly scattered around a central value that
provides the best estimate of a coin’s expected, or “true” mass. There are two common ways to estimate central tendency:
• mean
• median.

Note
When n = 5, the median is the third value in the ordered data set; for n = 6, the median is the average of the third and
fourth members of the ordered data set.
 Masses of Seven Circulating U. S. COINs
Coin Mass (g)
1 3.080
2 3.094
3 3.107
4 3.056
5 3.112
6 3.174
7 3.198

Calculate the mean & median for the data given in the above Table?
 Masses of Seven Circulating U. S. COINs
Coin Mass (g)
1 3.080
2 3.094
3 3.107
4 3.056
5 3.112
6 3.174
7 3.198

Calculate the mean (ans 3.117) & median (ans 3.107) for the data given in the above Table?
Measures of Spread

While mean or median provides an estimate of the coin’s expected mass, spread of individual measurements provides
an estimate of the difference in mass among coins or of the uncertainty in measuring mass with a balance.

• Although we often define Spread relative to a specific measure of central tendency, its magnitude is independent
of the central value. Changing all measurements in the same direction, by adding or subtracting a constant value,
changes the mean or median, but does not change the spread.
• There are three common measures of spread:
1) the range,
2) the standard deviation, and
3) the variance.
 Range
• The range, w, is the difference between a data set’s largest and smallest values.

• The range provides information about the total variability in the data set, but does not provide
any information about the distribution of individual values.

Standard Deviation
The standard deviation, s, describes the spread of a data set’s individual values about its mean, and is given as

the relative standard deviation, sr

Variance
Another common measure of spread is the square of the standard deviation, or the variance. We usually report a data set’s
standard deviation, rather than its variance, because the mean value and the standard deviation have the same unit.
Masses of Seven Circulating U. S. COINs

Coin Mass (g)

Calculate the range, standard deviation, the relative standard deviation, the percent relative
1 3.080
standard deviation and the variance for the data given in the above table?
2 3.094
3 3.107
4 3.056 1) Range:
5 3.112
6 3.174 2) standard deviation :
7 3.198

3) relative standard deviation and the percent relative standard deviation

4) variance:
problem
The following data were collected as part of a quality control study for the analysis of sodium in serum; results are
concentrations of Na+ in mmol/L.
140, 143, 141, 137, 132, 157, 143, 149, 118, 145

Report the mean, the median, the range, the standard deviation, and the variance for this data.
Characterizing Experimental Errors
Characterizing the mass of the COIN using the given data suggests two questions.

• First, does our measure of central tendency agree with the coin’s expected mass? → tells about the accuracy of
measurements

• Second, why is there so much variability in the individual results? → tells about precision

Two types of Errors are there:

1) Determinate Errors → errors affecting the 2) Indeterminate Errors → Errors affecting

accuracy of an analysis precision
Errors Affecting Accuracy - Determinate errors

Accuracy is a measure of how close a measure of central tendency is to the expected value, μ. We can express accuracy as
either an absolute error, e.

Determinate errors are of four categories:

1) sampling errors
2) method errors
3) measurement errors, and
4) personal errors
1-Sampling Errors

A determinate sampling error occurs when our sampling strategy does not provide a representative sample.
For example,
• if you monitor the environmental quality of a lake by sampling a single location near a point source of pollution,
such as an outlet for industrial effluent, then your results will be misleading.
• In determining the mass of a COIN, our strategy for selecting coins must ensure that we count only coins having
one value.

2-Method Errors
In any analysis the relationship between the total signal, Stotal and the absolute amount of analyte, nA, or the analyte’s
concentration, CA, is
Stotal = kAnA+Smb kA is the method’s sensitivity for the analyte and
Stotal = kACA+Smb Smb is the signal from the method blank.

A determinate method error exists when our value for kA or Smb is invalid
3-Measurement Errors
The manufacturers of analytical instruments and equipment, such as glassware and balances, usually provide a statement of the
item’s maximum measurement error, or tolerance.

For example, a 10-mL volumetric pipet has a tolerance of ±0.02 mL, which means
that the pipet delivers an actual volume within the range 9.98–10.02 mL at a
temperature of 20 oC. Although we express this tolerance as a range, the error is
determinate; thus, the pipet’s expected volume is a fixed value within the stated
range

❖ Volumetric glassware is categorized into classes depending on its accuracy.

• Class A glassware is manufactured to comply with tolerances specified by agencies such as the National Institute of
Standards and Technology or the American Society for Testing and Materials. The tolerance level for Class A glassware
is small enough that we normally can use it without calibration.
• Class B glassware tolerance levels are usually twice or more those for Class A glassware. Other types of volumetric
glassware, such as beakers and graduated cylinders, are unsuitable for accurately measuring volumes.
We can minimize determinate measurement errors by calibrating our equipment.
It is never safe to assume that a calibration will remain unchanged during an analysis or over time.
One study, for example, found that repeatedly exposing volumetric glassware to higher temperatures during machine washing
and oven drying, leads to small, but significant changes in the glassware’s calibration.
4- Personal Errors
Analytical work is always subject to personal error. You can minimize personal errors by taking proper care.

• including the ability to see a change in the color of an indicator signaling the endpoint of a titration;
• biases, such as consistently overestimating or underestimating the value on an instrument’s readout scale;
• failing to calibrate instrumentation; and misinterpreting procedural directions.
Errors Affecting Precision - Indeterminate Errors

Precision is a measure of the spread of individual measurements or results about a central value, which we express as a
range, a standard deviation, or a variance.

We make a distinction between two types of precision: repeatability and reproducibility.

• Repeatability is the precision when a single analyst completes the analysis in a single session using the same
solutions, equipment, and instrumentation.
• Reproducibility, on the other hand, is the precision under any other set of conditions, including between analysts, or
between laboratory sessions for a single analyst. Since reproducibility includes additional sources of variability, the
reproducibility of an analysis cannot be better than its repeatability.

Errors affecting precision are indeterminate and are characterized by random variations in their magnitude and their
direction. Because they are random, positive and negative indeterminate errors tend to cancel, provided that enough
measurements are made. In such situations the mean or median is largely unaffected by the precision of the analysis.
 Error vs Uncertainty

• Error is the difference between a single measurement • Uncertainty expresses the range of possible values
or result and its expected value. In other words, error for a measurement or result.
is a measure of bias. • Uncertainty accounts for all errors—both determinate
• Error can be divided into determinate and and indeterminate—that might reasonably affect a
indeterminate sources. measurement or result.
• Although we can correct for determinate errors, the • Although we always try to correct determinate errors
indeterminate portion of the error remains. before beginning an analysis, the correction itself is
subject to uncertainty.
Uncertainty vs Precision
Difference between precision and uncertainty:
Suppose you purchase a 10-mL Class A pipet from a laboratory supply company and use it without any additional calibration.
• The pipet’s tolerance of ±0.02 mL is its uncertainty because your best estimate of its expected volume is 10.00 mL ± 0.02
mL. This uncertainty is primarily determinate error.

• If you use the pipet to dispense several replicate portions of solution, the resulting standard deviation is the pipet’s
precision.

Experimental Results for Volume

Delivered by a 10-mL Class A Transfer
Pipet The table shows results for ten such trials, with a mean of 9.992 mL and a
Volume Volume standard deviation of ±0.006 mL. This standard deviation is the precision with
Number Number which we expect to deliver a solution using a Class A 10-mL pipet.
(mL) (mL)
1 10.002 6 9.983
2 9.993 7 9.991
In this case the published uncertainty for the pipet (±0.02 mL) is worse than its
3 9.984 8 9.990 experimentally determined precision (±0.006 ml).
4 9.996 9 9.988
5 9.989 10 9.999
Propagation of Uncertainty

Que - Suppose you dispense 20 mL of a reagent using the Class A 10-mL pipet whose calibration information is given in
the table. If the volume and uncertainty for one use of the pipet is 9.992 ± 0.006 mL (mean and standard deviation), what
is the volume and uncertainty when we use the pipet twice?

Experimental Results for Volume

Delivered by a 10-mL Class A Transfer
Pipet
Volume Volume
Number Number
(mL) (mL)
1 10.002 6 9.983
2 9.993 7 9.991
3 9.984 8 9.990
4 9.996 9 9.988
5 9.989 10 9.999
Ans
Experimental Results for Volume
• Mean = 9.992 mL and standard deviation = ±0.006 mL. Delivered by a 10-mL Class A Transfer
• As a first guess, we might simply add together the volume and the maximum Pipet
Volume Volume
uncertainty for each delivery; thus Number Number
(mL) (mL)
(9.992mL+9.992mL) ± (0.006mL+0.006mL) = 19.984 ± 0.012mL 1 10.002 6 9.983
2 9.993 7 9.991
3 9.984 8 9.990
Adding the uncertainty for the first delivery to that of the second delivery assumes
4 9.996 9 9.988
that with each use the indeterminate error is in the same direction and is as large 5 9.989 10 9.999
as possible.

• At the other extreme, we might assume that the uncertainty for one delivery is positive and the other is negative. If we
subtract the maximum uncertainties for each delivery,
(9.992mL+9.992mL) ± (0.006mL−0.006mL) = 19.984 ± 0.000mL
we clearly underestimate the total uncertainty.

So what is the total uncertainty ? From the above discussion we know that the total uncertainty is greater than ±0.000 mL
and less than ±0.012 mL. To estimate the cumulative effect of multiple uncertainties we use a mathematical technique
known as the propagation of uncertainty.
Propagation of Uncertainty – A few symbols

• A propagation of uncertainty allows us to estimate the uncertainty in a result from the uncertainties in the
measurements used to calculate the result.
For the equations in this section we represent the result with the symbol R, and the measurements with the symbols A, B,
and C. The corresponding uncertainties are uR, uA, uB, and uC.

We can define the uncertainties for A, B, and C using standard deviations, ranges, or tolerances (or any other measure of
uncertainty), as long as we use the same form for all measurements.
1. Uncertainty When Adding or Subtracting
When adding or subtracting measurements we use their absolute uncertainties for a propagation of uncertainty.
For example, if the result is given by the equation
R = A+B−C
then the absolute uncertainty in R is

2. Uncertainty When Multiplying or Dividing

When multiplying or dividing measurements we use their relative uncertainties for a propagation of uncertainty. For
example, if the result is given by the equation
R = (A × B) / C
then the relative uncertainty in R is
Uncertainty for Mixed Operations
Many chemical calculations involve a combination of adding and subtracting, and multiply and dividing.

We can calculate uncertainty by treating each operation separately using below equations
 Experimental Results for Volume Delivered by
Problems: a 10-mL Class A Transfer Pipet
Problem 1:
When dispensing 20 mL using a 10-mL Class A pipet, what is the total volume Volume Volume
Number Number
dispensed and what is the uncertainty in this volume? First, complete the (mL) (mL)
calculation using the manufacturer’s tolerance of 10.00 mL ± 0.02 mL, and 1 10.002 6 9.983
then using the calibration data from the table. 2 9.993 7 9.991
3 9.984 8 9.990
Problem 2: 4 9.996 9 9.988
5 9.989 10 9.999

Problem 3:
Problem 1:
When dispensing 20 mL using a 10-mL Class A pipet, what is the total volume dispensed and what is the uncertainty in this
volume? First, complete the calculation using the manufacturer’s tolerance of 10.00 mL ± 0.02 mL, and then using the calibration
data from the table.

Experimental Results for

Volume Delivered by a 10-
mL Class A Transfer Pipet
Number Volume
(mL)
1 10.002
2 9.993
3 9.984
4 9.996
5 9.989
6 9.983
7 9.991
8 9.990
9 9.988
10 9.999