Instrumental Analysis

INSTRUMENTAL METHODS OF ANALYSIS
Antonio G. Celajes Jr., RCh

WCC PRAYER
LAST MEETING’S LEARNING OBJECTIVES
At the end of the lesson, the students will be able to judge which titrimetric technique to be used when given a sample of known analyte.
a. Overview of Titrimetry
a.1 Equivalence Points and End Points
a.2 Volume as a Signal
a.3 Titration Curves
a.4 The Buret
b. Titrations Based on Acid–Base Reactions
b.1 Acid–Base Titration Curves
b.2 Selecting and Evaluating the End Point
b.3 Titrations in Non-aqueous Solvents
b.4 Representative Method
b.5 Quantitative Applications
b.6 Qualitative Applications
b.7 Characterization Applications
b.8 Evaluation of Acid–Base Titrimetry
c. Titrations Based on Complexation Reactions
c.1 Chemistry and Properties of EDTA
c.2 Complexometric EDTA Titration Curves
c.3 Selecting and Evaluating the End Point
c.4 Representative Method
c.5 Quantitative Applications
c.6 Evaluation of Complexation Titrimetry
d. Titrations Based on Redox Reactions
d.1 Redox Titration Curves
d.2 Selecting and Evaluating the End Point
d.3 Representative Method
d.4 Quantitative Applications
d.5 Evaluation of Redox Titrimetry
e. Precipitation Titrations
e.1 Titration Curves
e.2 Selecting and Evaluating the End Point
e.3 Quantitative Applications
e.4 Evaluation of Precipitation Titrimetry
TODAY’S LEARNING OBJECTIVES
At the end of the lesson, the students will be able to construct analytical procedures based on spectroscopic
methods of analysis.
Spectroscopic Method of Analysis

a. Overview of Spectroscopy
a.1 What Is Electromagnetic Radiation?
a.2 Measuring Photons as a Signal
b. Basic Components of Spectroscopic Instrumentation
b.1 Sources of Energy
b.2 Wavelength Selection
b.3 Detectors
b.4 Signal Processors
At the end of the lesson, the students will be able to construct analytical procedures based on
spectroscopic methods of analysis.
c. Spectroscopy Based on Absorption

c.1 Absorbance of Electromagnetic Radiation
c.2 Transmittance and Absorbance
c.3 Absorbance and Concentration: Beer’s Law
c.4 Beer’s Law and Multicomponent Samples
c.5 Limitations to Beer’s Law
At the end of the lesson, the students will be able to construct analytical procedures based on spectroscopic
methods of analysis.
d. Ultraviolet-Visible and Infrared Spectrophotometry

d.1 Instrumentation
d.2 Quantitative Applications
d.3 Qualitative Applications
d.4 Characterization Applications
d.5 Evaluation
e. Atomic Absorption Spectroscopy
e.1 Instrumentation
e.2 Quantitative Applications
e.3 Evaluation
Instrumental analysis is a field of analytical chemistry that investigates analytes using scientific
instruments.
An instrument is a communication device between the system under study and the investigator.
Instrument converts
Information contained in Information for human
Chemical and physical →→→→→→→→ interpretation in terms
Characteristics of property of interest and
manipulation
SPECTROSCOPIC METHOD OF ANALYSIS
Spectroscopy
Spectroscopy and spectrography are terms used to refer to the measurement of radiation
intensity as a function of wavelength and are often used to describe experimental spectroscopic
methods.
Spectral measurement devices are referred to as spectrometers, spectrophotometers,
spectrographs or spectral analyzers.
Electromagnetic radiation—light
Form of energy whose behavior is described by the properties of both waves and particles;
Some properties of electromagnetic radiation, such as its refraction when it passes from one
medium to another are explained best by describing light as a wave;
Other properties, such as absorption and emission, are better described by treating light as a
particle.
Wave Properties of Electromagnetic Radiation
Electromagnetic radiation consists of oscillating electric and magnetic fields that
propagate through space along a linear path and with a constant velocity;
In a vacuum electromagnetic radiation travels at the speed of light, c, which is 2.99792
× 108 m/s.
When electromagnetic radiation moves through a medium other than a vacuum its velocity,
v, is less than the speed of light in a vacuum;
The difference between v and c is sufficiently small (<0.1%) that the speed of light to
three significant figures, 3.00 × 108 m/s, is accurate enough for most purposes.
An electromagnetic wave is characterized by several fundamental properties, including its
velocity, amplitude, frequency, phase angle, polarization, and direction of propagation.
Wave Properties of Electromagnetic Radiation
The amplitude of a wave is a measure of the displacement of
the wave from its rest position;
Frequency is the number of occurrences of a repeating event
per unit of time.
The wavelength, λ, is defined as the distance between

successive maxima
λ=c/v where c = speed of light v = frequency
Wavenumber is the reciprocal of wavelength v = 1/λ

Particle Properties of Electromagnetic Radiation
When matter absorbs electromagnetic radiation it undergoes a change in energy;
The interaction between matter and electromagnetic radiation is easiest to understand if we
assume that radiation consists of a beam of energetic particles called photons;
When a photon is absorbed by a sample it is “destroyed,” and its energy acquired by the
sample;
The energy of a photon, in joules, is related to its frequency, wavelength, and wavenumber by
the following equalities
where E = energy; h = 6.63 x 𝟏𝟎−𝟑𝟒 Js (Planck’s constant); v = frequency;

c = 3.00 x 𝟏𝟎𝟖 m/s
SPECTROSCOPY BASED ON ABSORPTION
Sample Problems
1. At 700nm, how much energy does the red of the visible spectrum possess?
I. Given: λ = 700nm = 7.00 x 𝟏𝟎−𝟕 m
II. Required: E
III. Solution:
hc (6.63 x 𝟏𝟎 −𝟑𝟒 Js) (3.00 x 𝟏𝟎𝟖 m/s)

E = ------ = -----------------------------------------
λ 7.00 x 𝟏𝟎−𝟕 m
c = 3.00 x 𝟏𝟎𝟖 m/s
= 2.84 x 𝟏𝟎−𝟏𝟗 J
h = 6.63 x 𝟏𝟎−𝟑𝟒 Js
Sample Problems
1. What would have been the wavelength of an electromagnetic radiation if its energy is 3.78
x 10−19 J?
I. Given: E = 3.78 x 𝟏𝟎−𝟏𝟗 J
II. Required: λ
III. Solution:
hc hc (6.63 x 𝟏𝟎 −𝟑𝟒 Js) (3.00 x 𝟏𝟎𝟖 m/s)

E = ------ λ = ------ = -----------------------------------------
λ E 3.78 x 𝟏𝟎−𝟏𝟗 J
c = 3.00 x 𝟏𝟎𝟖 m/s Eλ = hc
----- -----
E E = 5.26 x 𝟏𝟎−𝟕 m = 526 nm
h = 6.63 x 𝟏𝟎−𝟑𝟒 Js
The Electromagnetic Spectrum
The frequency and wavelength of
electromagnetic radiation vary over
many orders of magnitude;
For convenience, we divide
electromagnetic radiation into different
regions—the electromagnetic spectrum—
based on the type of atomic or molecular
transition that gives rise to the absorption
or emission of photons;
The boundaries between the regions of
the electromagnetic spectrum are not
rigid, and overlap between spectral
regions is possible.
Photons as a Signal Source
A spectroscopic measurement is possible only if the photon’s interaction with the sample
leads to a change in one or more of these characteristic properties;
We can divide spectroscopy into two broad classes of techniques
1. Spectroscopic Techniques Involving an Exchange of Energy Between a Photon and the
Sample and
2. Spectroscopic Techniques That Do Not Involve an Exchange of Energy Between a
Photon and the Sample
Spectroscopic Techniques Involving an Exchange of Energy Between a Photon and the Sample
Spectroscopic Techniques That Do Not Involve an Exchange of Energy Between a Photon and the
Sample
Basic Components of Spectroscopic Instruments
Sources of Energy
A source of electromagnetic radiation must provide an output that is both intense and
stable;
Sources of electromagnetic radiation are classified as either continuum or line sources;
A continuum source emits radiation over a broad range of wavelengths, with a relatively
smooth variation in intensity;
A line source, on the other hand, emits radiation at selected wavelengths.
Basic Components of Spectroscopic Instruments Sources of Energy
Sources of Thermal Energy
The most common sources of thermal energy are flames and plasmas;
Flames sources use the combustion of a fuel and an oxidant to achieve temperatures of
2000–3400 K;
Plasmas, which are hot, ionized gases, provide temperatures of 6000–10 000 K.
Chemical Sources of Energy
Exothermic reactions also may serve as a source of energy. In chemiluminescence the
analyte is raised to a higher-energy state by means of a chemical reaction, emitting
characteristic radiation when it returns to a lower-energy state;
When the chemical reaction results from a biological or enzymatic reaction, the emission
of radiation is called bioluminescence. Commercially available “light sticks” and the
flash of light from a firefly are examples of chemiluminescence and bioluminescence.
Wavelength Selection
The ideal wavelength selector has a high throughput of radiation and a narrow effective
bandwidth;
A high throughput is desirable because more photons pass through the wavelength
selector, giving a stronger signal with less background noise;
A narrow effective bandwidth provides a higher resolution, with spectral features
separated by more than twice the effective bandwidth being resolved.
Wavelength Selection Using Filters
The simplest method for isolating a narrow band of radiation is to use an absorption or
interference filter;
Absorption filters work by selectively absorbing radiation from a narrow region of the
electromagnetic spectrum;
A simple example of an absorption filter is a piece of colored glass;
Interference filters use constructive and destructive interference to isolate a narrow
range of wavelengths.
Wavelength Selection Using Monochromators
A filter has one significant limitation—because a filter has a fixed nominal wavelength,
if you need to make measurements at two different wavelengths, then you need to use
two different filters;
A monochromator is an alternative method for selecting a narrow band of radiation that
also allows us to continuously adjust the band’s nominal wavelength;
Radiation exits the monochromator and passes to the detector;
Monochromator converts a polychromatic source of radiation at the entrance slit to a
monochromatic source of finite effective bandwidth at the exit slit.
Wavelength Selection Using Monochromators
Monochromators are classified as either fixed-wavelength or scanning;
In a fixed-wavelength monochromator we select the wavelength by manually rotating the
grating;
Normally a fixed-wavelength monochromator is used for a quantitative analysis where
measurements are made at one or two wavelengths;
A scanning monochromator includes a drive mechanism that continuously rotates the grating,
allowing successive wavelengths to exit from the monochromator;
Scanning monochromators are used to acquire spectra, and, when operated in a fixed-
wavelength mode, for a quantitative analysis.
Interferometers
An interferometer provides an alternative approach for wavelength selection;
Instead of filtering or dispersing the electromagnetic radiation, an interferometer allows
source radiation of all wavelengths to reach the detector simultaneously;
Radiation from the source is focused on a beam splitter that reflects half of the radiation
to a fixed mirror and transmits the other half to a movable mirror;
The radiation recombines at the beam splitter, where constructive and destructive
interference determines, for each wavelength, the intensity of light reaching the detector.
Detectors
The human eye, of course, has a poor range—responding only to visible light—nor is it
particularly sensitive or accurate;
Modern detectors use a sensitive transducer to convert a signal consisting of photons into
an easily measured electrical signal.
Transducer is a general term that refers to any device that converts a chemical or
physical property into an easily measured electrical signal;
The retina in your eye, for example, is a transducer that converts photons into an
electrical nerve impulse;
There are two broad classes of spectroscopic transducers: thermal transducers and
photon transducers.
Detectors
Photon Transducers.
Phototubes and photomultipliers contain a photosensitive surface that absorbs radiation in the
ultraviolet, visible, or near IR, producing an electrical current proportional to the number of
photons reaching the transducer;
Thermal Transducers.
Infrared photons do not have enough energy to produce a measurable current with a photon
transducer;
A thermal transducer, therefore, is used for infrared spectroscopy;
The absorption of infrared photons by a thermal transducer increases its temperature, changing
one or more of its characteristic properties.
Signal Processors
A transducer’s electrical signal is sent to a signal processor where it is displayed in a
form that is more convenient for the analyst;
Examples of signal processors include analog or digital meters, recorders, and computers
equipped with digital acquisition boards;
A signal processor also is used to calibrate the detector’s response, to amplify the
transducer’s signal, to remove noise by filtering, or to mathematically transform the
signal.
Absorbance of Electromagnetic Radiation
In absorption spectroscopy a beam of electromagnetic radiation passes through a sample;
Much of the radiation is transmitted without a loss in intensity;
At selected frequencies, however, the radiation’s intensity is attenuated;
This process of attenuation is called absorption;
Two general requirements must be met if an analyte is to absorb electromagnetic radiation;
The first requirement is that there must be a mechanism by which the radiation’s electric field or
magnetic field interacts with the analyte.
For ultraviolet and visible radiation, this interaction involves the electronic energy of valence
electrons. A chemical bond’s vibrational energy is altered by the absorbance of infrared
radiation.
Absorbance of Electromagnetic Radiation
The second requirement is that the energy of the electromagnetic radiation must exactly
equal the difference in energy, ∆E, between two of the analytes quantized energy
states.
The bonds and functional groups that give rise to the absorption of ultraviolet and visible
radiation are called chromophores.
Transmittance and Absorbance
As light passes through a sample, its power decreases as some of it is absorbed;
This attenuation of radiation is described quantitatively by two separate, but related terms:
transmittance and absorbance.
Transmittance is the ratio of the source radiation’s power exiting the sample, PT, to that incident
on the sample, P0.
T = PT / P0 or T = I / I0 Where I0 is the intensity of the incident light beam and I is the
intensity of the light coming out of the sample.
Multiplying the transmittance by 100 gives the percent transmittance, %T, which varies between
100% (no absorption) and 0% (complete absorption);
All methods of detecting photons—including the human eye and modern photoelectric
transducers—measure the transmittance of electromagnetic radiation.
Transmittance and Absorbance
An alternative method for expressing the attenuation of electromagnetic radiation is
absorbance, A , which we define as
Absorbance is the more common unit for expressing the attenuation of radiation because
it is a linear function of the analyte’s concentration.
Absorbance and Concentration: Beer’s Law
A = εbC where A = absorption; ε = molar absorptivity(/cmM);

b = cell pathlength; C = molar concentration
Limitations to Beer’s Law
Fundamental Limitations to Beer’s Law
Beer’s law is a limiting law that is valid only for low concentrations of analyte. There are
two contributions to this fundamental limitation to Beer’s law.
1. At higher concentrations the individual particles of analyte no longer behave

independently of each other. The resulting interaction between particles of analyte may
change the analyte’s absorptivity.
2. A second contribution is that the analyte’s absorptivity depends on the sample’s
refractive index. Because the refractive index varies with the analyte’s concentration, the
values of a and ε may change. For sufficiently low concentrations of analyte, the
refractive index is essentially constant and the calibration curve is linear.
QUANTITATIVE APPLICATIONS
Sample Problems
1. What would have been the %transmittance of the radiation in the sample if the intensity of
the light before entering the sample is 2.56cd while the light that came out of the sample is
0.77cd?
I. Given: Io = 2.56cd I = 0.77cd
II. Required: %T
III. Solution:
I I 0.77cd
T = ------ %T = ------ x 100 = ---------- x 100 = 30.08%
Io Io 2.56cd
Sample Problems
2. A sample was recorded to have %transmittance of 67.89%. What would have its molar
concentration if it was placed in a 1.50cm cell? The molar absorptivity of the sample is
548/cmM?
I. Given: %T = 67.89% T = 67.89/100 = 0.6789 b = 1.50cm ε = 548/cmM
II. Required: M
III. Solution:
0.1682
A = εbC C = ----------------------------
------ ------ (548 / cmM)(1.50cm)
εb εb
A A = -logT = -log0.6789
C = ------ = 0.000205M
εb = 0.1682
= 2.05 x 𝟏𝟎−𝟒 𝐌
Sample Problems
3. The concentration of a solution of NaCl was analyzed using UV-Vis spectroscopy. A
transmittance of 0.637 was recorded using a 2.40cm cell. How many grams of NaCl in a liter
solution must have been used if its molar absorptivity was found to be 628/cmM? Na= 23; Cl
=35
I. Given: T = 0.637 b = 2.40cm V = 1.00L ε = 628/cmM
MW NaCl = (1)(23) + (1)(35) = 58g/mole
II. Required: wt of NaCl in g
III. Solution:
wt = (M)(MW)(V in L) A = -logT = -log0.637 = 0.1959 wt = (0.000130mole/L)(58g/mol)(1 L)

A 0.1959 = 0.0075g
C = ------ = ---------------------------- = 0.000130M = 1.30 x 𝟏𝟎−𝟒 𝐌
εb (628 / cmM)(2.40cm)

Instrumental Analysis

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INSTRUMENTAL METHODS OF ANALYSIS

Antonio G. Celajes Jr., RCh

Spectroscopic Method of Analysis

c. Spectroscopy Based on Absorption

d. Ultraviolet-Visible and Infrared Spectrophotometry

The wavelength, λ, is defined as the distance between

Wavenumber is the reciprocal of wavelength v = 1/λ

where E = energy; h = 6.63 x 𝟏𝟎−𝟑𝟒 Js (Planck’s constant); v = frequency;

hc (6.63 x 𝟏𝟎 −𝟑𝟒 Js) (3.00 x 𝟏𝟎𝟖 m/s)

hc hc (6.63 x 𝟏𝟎 −𝟑𝟒 Js) (3.00 x 𝟏𝟎𝟖 m/s)

A = εbC where A = absorption; ε = molar absorptivity(/cmM);

1. At higher concentrations the individual particles of analyte no longer behave

wt = (M)(MW)(V in L) A = -logT = -log0.637 = 0.1959 wt = (0.000130mole/L)(58g/mol)(1 L)

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