Cebu Doctors’ University

College of Arts and Sciences

Physical Sciences Department

MODULE 1.1 – INTRODUCTION TO ANALYTICAL CHEMISTRY

Compiled by: Joselito R. Tumulak Jr., RChT, MS (cand.)
Analytical Chemistry Professor

INTENDED LEARNING OUTCOMES

At the end of this module, you will be able to:
1. Define the scope of analytical chemistry and its role in other sciences, especially in clinical
laboratories;
2. Contrast different quantitative analytical methods; and
3. Explain each step in a typical chemical analysis.

UNIT OUTLINE
Topic Page
I. Scope and Nature of Analytical Chemistry
A. Definition of Analytical Chemistry 1
B. Applications of Analytical Chemistry
II. Methods of Analysis
A. Levels of Analytical Methodology 6
B. Quantitative Analytical Methods
III. Flow of Chemical Analysis 16

I. SCOPE AND NATURE OF ANALYTCAL CHEMISTRY

A. Definition of Analytical Chemistry

 Analytical chemistry is often defined as the science of obtaining, separating, identifying, and
quantifying matter.
 Some important terms that often used in analytical chemistry:
- Sample – substances to be analyzed.
- Analyte – components of the samples to be determined/detected.
 For example, a blood sample is analyzed to determine the concentrations of various
substances, such as blood gases. The sample is blood, while the analytes are the
blood gases. We, therefore, speak of the determination of blood gases, not the
analysis of blood gases or glucose.
- Matrix – remainder of the sample other than the analyte.

 Although analytical chemistry is related to chemical analysis, remember that the craft of analytical
chemistry is not in performing a routine chemical analysis, but in improving established
methods, extending existing methods to new types of samples, and developing new methods
for measuring chemical phenomena.
- To help us understand this, consider this example. Mining engineers evaluate the economic
feasibility of extracting an ore by comparing the cost of removing the ore with the value of its
contents. To estimate its value, they analyze a sample of the ore. The challenge of
developing and validating the method providing this information is the analytical chemist’s
responsibility. Once developed, the routine, daily application of the method becomes the job
of the chemical analyst.

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  Types of Analysis in Analytical Chemistry:
- Characterization analysis – evaluates a sample’s chemical or physical properties.
Examples are determination of chemical structure, equilibrium constants, particle size, and
surface structure.
 Characterization analysis may be:
1. Qualitative analysis – establishes the chemical identity of species in the
sample (detection/identification)
2. Quantitative analysis – determines the relative amounts of the different
species in numerical terms (determination)
- Fundamental analysis - directed toward improving analytical method’s capabilities.
Examples are extending and improving the theory on which a method is based, studying a
method’s limitations, and designing new and modifying old methods.

Examples:

Consider the following situation:

1. A chemistry student prepared an ethanolic extract of malunggay leaves and wanted to determine
the types of polyphenols present in the extract. Using high-performance liquid chromatography
(HPLC), the student confirmed the presence of myrecytin, quercetin, kaempferol, gallic acid,
chlorogenic acid, and caffeic acid.

a. What is the sample in the given situation?

b. What is the analyte?
c. What form of analysis was done?

2. A student researcher obtained water samples from deep wells in a coastal barangay to determine
the extent of seawater intrusion. To determine the chloride ion concentration, the student used a
specific ion electrode for chlorides. The voltage reading obtained from the instrument was used to
calculate the concentration of chlorides in terms of mg/L.

a. What is the sample in the given situation?

b. What is the analyte?
c. What form of analysis was done?

If you answered:
1. a. ethanolic extract of malunggay leaves
b. polyphenols (myrecytin, quercetin, kaempferol,
gallic acid, chlorogenic acid, and caffeic acid)
c. Characterization Analysis - Qualitative analysis

and

2. a. water from deep wells

b. chloride ions
c. Characterization Analysis - Quantitative analysis

B. Application of Analytical Chemistry

 To illustrate the central role of analytical chemistry in other sciences, consider some example below:
- Clinical Chemistry
1. Determination the concentration of different substances in urine can be used to
diagnose and treat illness.
2. Quantitative measurements of calcium ion in blood serum help diagnose parathyroid
disease in humans

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 3. Determination of potassium and sodium ions in the body fluids permits the study of
these ions play in nerve-signal conduction, as well as muscle contraction and
relaxation.
- Material Science
1. Analysis of steel during its production allows adjustment in the concentrations of such
elements as carbon, nickel, and chromium to achieve a desired strength, hardness,
corrosion resistance, and ductility.
2. Quantitative analysis of crystalline germanium and silicon ensures that impurities are
controlled in semiconductor devices.
- Food Science
1. Quantitative determination of nitrogen in foods establishes their protein content and
thus their nutritional value.
2. Control of the amount of harmful preservatives and additives in processed food is
maintained by doing rigid chemical analysis.
- Environmental Chemistry
1. Quantities of hydrocarbons, nitrogen oxides, and carbon monoxide present in
automobile exhaust gases are measured to determine the effectiveness of emission-
control devices.
2. Pollutants in the river, such as heavy metals and organic wastes, are monitored
through quantitative analysis of these compounds.

II. ANALYTICAL METHODS

A. Levels of Analytical Methodology

 There four levels of analytical methodology:

1. Techniques – a chemical or physical principle that can be used to analyze a sample
2. Methods – the application of a technique for the determination of a specific analyte in a
specific matrix.
3. Procedures – a set of written directions detailing how to apply a method to a particular
sample, including information on proper sampling, handling of interferents, and validating
results
4. Protocols – a set of stringent written guidelines detailing the procedure that must be
followed if the agency specifying the protocol is to accept the results of the analysis.
Protocols are commonly encountered when analytical chemistry is used to support or define
public policy.

 To illustrate these four levels, consider a situation when you are asked to develop a way to determine
the concentration of lead in drinking water. How would you approach this problem?

Technique Many techniques have been used to determine lead levels. For example, in atomic
absorption spectroscopy lead is atomized, and the ability of the free atoms to absorb
light is measured; thus, both chemical principle (atomization) and a physical
principle (absorption of light) are used in this technique.
Method Atomic absorption spectroscopic method for determining lead levels in water is
different from that for the determination of lead in soil or blood. Choosing a method
for determining lead in water depends on how the information is to be used and the
established design criteria.
Procedure A method does not necessarily lead to a single procedure, as different analysts or
agencies will adapt the method to their specific needs. For example, the American
Public Health Agency and the American Society for Testing Materials publish
separate procedures for the determination of lead levels in water.
Protocol For purposes of determining lead levels in water under the Safe Drinking Water Act,
labs follow a protocol specified by the Environmental Protection Agency.
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B. Quantitative Analytical Methods

 Analytical chemistry in the modern times devote much time in the laboratory gathering quantitative
information about systems. Qualitative analysis is often part of an analytical procedure as a
preliminary test.
 The various methods of quantitative analysis are classified according to the measurable property of
the analyte that can be used to calculate the concentration of the analyte.
 Quantitative analytical methods can be classified as:
- Classical Methods – also known as wet methods or batch analysis and is achieved by
measurement of weight or volume. Calculations are often derived from stoichiometry of a
chemical reaction. Classical methods include:
o Gravimetry - determines mass of the analyte or some compound chemically related
to it. A typical gravimetric analysis involves precipitation, filtration, drying and
weighing of the precipitate.
- Example:
The amount of calcium in a solution can be determined by gravimetric
analysis using oxalate ion as precipitating agent.

Step 1
Dissolve Step 3
Filter, dry, and
weigh precipitate as
calcium oxalate
Step 2
Add solution
Sample containing
containing oxalate
calcium

o Titrimetry – also known as volumetric analysis; the volume of a solution containing

sufficient reagent to react completely with the analyte is measured. The important
used in titration is the buret.
- Example:
The acidity of vinegar is measured by titrating with a standard sodium
hydroxide solution.

- Instrumental Methods – uses instrumentations to measure physical properties of analyte,

such as current, potential, and absorption or emission of light. These physical properties are
directly proportional to the concentration of the analyte in the sample. The instrumental
methods of analysis are divided into categories according to the property of the analyte that
is to be measured. Instrumental methods of analysis can be:

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 o Spectroscopic methods – based on measurements of the interaction between
electromagnetic radiation and analyte atoms or molecules, or on the production of
such radiation by analytes.
- Some important terminologies:
o Spectroscopy – study of interaction of radiation and matter
o Spectrophotometry – measurement of emission or absorption of
radiation from matter
o Spectrophotometer – instrument used to measure emission or
absorption of radiation from matter
o Spectrum – the output of a spectrophotometer
- A Typical Spectrophotometer:

- The Electromagnetic Radiation:

- Different EM radiation will interaction with matter differently; thus, different

spectroscopic methods are based on the type of radiation used. Common
spectroscopic methods include:
o UV – Visible Spectrophotometry – uses radiation in the UV and
Visible region; results in electronic excitation in molecules. This is
often used in analyzing analytes that are colored or very rich double
bonds (conjugated).
o Infrared Spectrophotometry – uses infrared radiation which results
to the vibration of the chemical bonds in molecules. This is applied in
identification of functional groups in organic compounds.
o NMR Spectrophotometry – uses radio waves which results to the
alignment of the nuclei of the atoms in molecules. This is applied in
determining the alignment of elements in a molecule, and therefore its
structure.

o Electroanalytical Methods – involve the measurement of properties such as

potential difference, current, resistance, and conductivity. Electrodes are commonly
used in these methods.

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 - Different electroanalytical methods are distinguished from each other based
on the type of property being measured:
o Potentiometry – the difference in electrode potentials is measured
o Amperometry – the electric current is measured
o Coulometry – the charge passed during a certain time is recorded
o Voltammetry – the current is measured while actively altering the
potential.

o Chromatographic Methods – involves the separation of the components of a

mixture based on differences in the rates at which they are carried through a fixed
stationary phase by a gaseous or liquid mobile phase.

High-performance Liquid Chromatography Gas Chromatography

- Chromatographic methods can be classified as:
o High-Performance Liquid Chromatography – liquid mobile phase
and the sample must be soluble in a selected solvent.
o Gas Chromatography – gaseous mobile phase and the sample must
be gaseous, volatile, or can be converted into volatile derivative.
- In essence, chromatography is only used to separate the analytes in a sample
for better selectivity. This separation takes place in the column. Other
methods, such as spectroscopic or electroanalytical methods, are used to
quantify the separated analytes. The detector performs this quantification,
which is then processed by a computer.

Examples:

Identify the method of analysis used in the given conditions.

1. Rate of photosynthesis in cassava leaves at different wavelengths of light were determined based
from the absorbance of the leaves extract.
2. Titration analysis of oxalic acid in fruit extracts of starfruits.
3. Percent (%) Cu in ore was determined based on the mass of Cu isolated from the ore.
4. The components of a body fluid were separated and identified based on their rate of diffusion along
a SiO2 column.
5. The mg/L of dissolved salts in a water sample was assessed by measuring the samples’ potential
difference.
6. The alkalinity of a soap solution was determined based on the milliliters of the acid needed to
neutralize the soap solution.

If you answered:
1. Spectroscopic method
2. Titrimetry
3. Gravimetry
4. Chromatographic method
5. Electroanalytical method
6. Titrimetry
Then, you are correct!

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  Advantages and Disadvantages of classical methods:
Advantages Disadvantages
1. Established and widely used, providing 1. Limited sensitivity and specificity.
well-understood results. 2. May not be suitable for trace or difficult-
2. Inexpensive and widely accessible. to-detect compounds.
3. Simple to perform and interpret. 3. Can be time-consuming and labor-
4. Often provides quick results. intensive.
4. Can suffer from interference from other
compounds.
5. Results can be influenced by operator
skill and laboratory conditions.

 Advantages and Disadvantages of instrumental methods:

Advantages Disadvantages
1. High sensitivity and specificity. 1. More expensive than classical methods.
2. Ability to detect trace or difficult-to- 2. More complex to perform and interpret.
detect compounds. 3. Requires specialized training and
3. Increased speed and automation equipment.
compared to classical methods. 4. Can suffer from interference from other
4. Improved precision and accuracy. compounds.
5. Minimal operator dependence. 5. Can be affected by instrument
maintenance and drift.

III. FLOW OF CHEMICAL ANALYSIS

 A typical quantitative analysis includes the sequence of steps shown in the flow diagram below. In
some instances, one or more of these steps can be omitted.

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 1. Selecting a method.
- This step usually requires experience and intuition.
- Requirements in selecting a method:
o level of accuracy and precision
o time and available resources
o number of available samples
o complexity of sample and number of components to be analyzed
- Chemical analysts often look for methods of analyzing different samples in published
methods from reliable organization such as Association of Official Analytical Chemists
(AOAC) and United States Pharmacopeia (USP).
- For example, in pharmaceuticals, if you want to determine the purity of a specific drug
formulation, such as fever medicine containing paracetamol, you can use the USP method.
This method has been validated by numerous independent laboratories and experts around
the world.
-
2. Acquiring the sample.
- Analysis must be performed on a sample that has the same composition as the bulk material
from which it was taken.
- Sampling is the process of collecting a small mass of a material whose composition
accurately represents the bulk of the material being sampled.
- Consideration on Sampling:
o The sample is collected in a manner consistent with the goal of analysis.
o The sample should be handled so as to prevent its contamination or alteration
o The sample is brought to the laboratory and prepared for the technique chosen.
- The majority of published methods include an established sampling method to be used for
analysis.
- Sampling is frequently the most difficult step in an analysis and the source of greatest error.
The final analytical result will never be any more reliable than the reliability of the sampling
step.
- For example, sampling of human blood for determination of blood gases must follow strict
procedures. This is because the concentration of oxygen and carbon dioxide in the blood
depends on variety of physiological and environment variables, such as inappropriate
application of tourniquet.

3. Processing the Sample

- After acquiring the sample, the sample must be processed accordingly for analysis in the
laboratory.
- Preparing a Laboratory Sample
o A solid laboratory sample is ground to decrease particle size, mixed to ensure
homogeneity, and stored for various lengths of time before analysis begins.
o For liquid samples, evaporation of solvent must be prevented which may change
concentration of the analyte.
o If the analyte is a gas dissolved in a liquid, as in our blood gas example, the sample
container must be kept inside a second sealed container, perhaps during the entire
analytical procedure, to prevent contamination by atmospheric gases.
- Defining Replicates
o A replicate is a portion of a material of approximately the same size that are carried
through an analytical procedure at the same time and in the same way. The mass or
volume of a replicate must be determined by careful measurements with an analytical
balance or with a precise volumetric device.
o Replication improves the quality of the results and provides a measure of their
reliability.
o Quantitative measurements on replicates are usually averaged, and various
statistical tests are performed on the results to establish their reliability.
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 - Dissolution of sample
o Most analyses are performed on solutions of the sample made with a suitable
solvent.
o Choose an appropriate solvent to dissolve the entire sample.
o Harsh chemistry may be involved to convert the analyte into a soluble form, such as
heating with aqueous solutions of strong acids, strong bases, oxidizing agents,
reducing agents, or some combination of such reagents.

4. Eliminating Interferences
- An interference is a species that causes an error in an analysis by enhancing or attenuating
(making smaller) the quantity being measured. An interference may share similar chemical
or physical properties that is important in the analysis of an analyte.
- A scheme must be devised to isolate the analytes from interferences before the final
measurement is made. No hard and fast rules can be given for eliminating interference.
- For example, the sensitivity of calcium measurements is reduced by the presence of
aluminum, silicon, phosphate and sulfate. Ethylenediaminetetraacetic acid (EDTA)
complexes with the calcium and eliminates these interferences.

5. Measuring the Physical or Chemical Property

- In quantitative analysis, we quantity a property that is proportional to the amount of analyte
in the sample such as mass, volume, intensity of light, or electrical charge.
- Again, the property to be measured is dependent on the type of method.
- For example, concentration of hexavalent chromium (Cr6+) in water is determined by reacting
Cr6+ with 1,5-diphenylcarbazide (DPC) dye forming a purple-colored species. The intensity
of color produced after the reaction is directly proportional to the Cr 6+ present. The intensity
of color is quantitatively measured as light absorption in a spectrophotometer.

6. Calculating the Results

- Calculating analyte concentrations are based on
the raw experimental data collected in the
measurement step, characteristics of the
measurement instrument, and the stoichiometry of
the analytical reaction.
- If the procedure uses classical methods,
calculations is based on stoichiometry of the
reaction. However, if the procedure uses
instrumental methods, calculations is based on a
calibration curve plot (standard curve plot) just
like the one on the right.

7. Estimating the Reliability of Results

- The chemical analyst must provide some measure
of the uncertainties associated with the calculated results.
- An analytical result without an estimate of reliability is of no value.

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