Instrumental Technique for Environmental Analysis Chapter 2 Spectroscopy

Rezaul Karim Environmental Science and Technology Jessore Science and Technology University

Principles of Spectroscopy
Electromagnetic radiation; general and wave properties; Electromagnetic spectrum; Interaction of light with matter; quantum mechanical properties of radiation; quantitative aspects of Spectro-chemical measurements; Optical Systems Used in Spectroscopy

Reference

Francis Rouessac and Annick Rouessac, 2007, Chemical Analysis-Modern Instrumentation Methods and Techniques, 2nd Edition, John Wiley & Sons Ltd, England James W Robinson, 1995. Undergraduate instrumental analysis, Marcel Dekker, Inc. NY Skoog, Holler & Crouch 2007, Instrumental Analysis, Brooks Cole Cengage Learning, USA. Daniel C. Harris , 2010, Quantitative Chemical Analysis , 8th edition, W. H. Freeman and Company 41 Madison Avenue New York.
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What is spectroscopy?
Spectroscopy is the science that deals with the interactions of various types of radiation with matter. Spectrometry and spectrometric methods refer to the measurement of the intensity of radiation with a photoelectric transducer or other type of electronic device. It is based on atomic and molecular spectroscopy. The most widely used spectrometric methods are based on electromagnetic radiation (EMR).

Processes in Spectroscopy
Absorption: The process by which the energy of the light is transferred to the atom or molecule raising them from the ground state to an excited state Fluorescence: The absorbed energy is rapidly lost to the surroundings by collisions within the system and relax back to the ground state. Emission: If the substances (atoms or molecules) are heated to high temperatures (in a flame or in an electric discharge) the electrons are exited to higher energy levels. Later, they relax to the ground state with the emission of radiation.

What is Light?
Light is a form of energy Light travels through space at extremely high velocities

The speed of light (c) ~ 3 x 1010 cm/sec or 186,000 miles per second

Light is classified as electromagnetic radiation (EMR)

Electromagnetic radiation

EMR is a type of energy that takes several forms e.g. light and radiant heat. The properties of electromagnetic radiation are conveniently described by means of a classical sinusoidal wave model which embodies such characteristics as wavelength, frequency, velocity, and amplitude. In contrast to other wave phenomena, such as sound, electromagnetic radiation requires no supporting medium for its transmission and thus passes readily through a vacuum. The energy of a photon is proportional to the frequency of the radiation.
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Sinusoidal wave model

electric and magnetic fields that undergo in-phase, sinusoidal oscillations at right angles to each other and to the direction of propagation.

The abscissa of this plot is either time as the radiation passes a fixed point in space or distance when time is held constant. In this figure, the electric field is in the xy plane, and the magnetic field is in the xz plane.

Wave properties

The time in seconds required for the passage of successive maxima or minima through a fixed point in space is called the period p of the radiation.

Wavelength (l) is the linear distance between any two equivalent points on successive waves (e.g., successive maxima or minima). Other way, l, is the crest-to-crest distance between waves. Amplitude is the vertical distance from the midline of a wave to the peak or trough. Frequency (v) is the number of waves that pass through a particular point in 1 second (Hz = 1 cycle/s) 106 s-1 = 106 Hz or 1 MHz

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Wave number , which is defined as the reciprocal of the wavelength in centimeters, The unit for is cm-1 it is directly proportional to the frequency, and thus the energy, of radiation. Thus, we can write, = kv where the proportionality constant k depends on the medium and is equal to the reciprocal of the velocity
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The power P of radiation is the energy of the beam that reaches a given area per second, The intensity I is the power per unit solid angle. These quantities are related to the square of the amplitude A . The energy of a photon depends on its frequency (v)

Ephoton = hv

h = Plancks constant h = 6.63 x 10-27 erg sec or 6.63 x 10-34 Js

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V = Wave Number (cm )

-1

l = Wave Length
C = Velocity of Radiation = 3 x 1010 cm/sec. u = Frequency of Radiation (cycles/sec)

The relation between frequency and wavelength is

V = u C = l

E = hu= h

C l
u = C l

u =

C l

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By how many kilojoules per mole is the energy of O2 increased when it absorbs ultraviolet radiation with a wavelength of 147 nm? How much is the energy of CO2 increased when it absorbs infrared radiation with a wavenumber of 2300 cm-1?

Try yourself: What is the wavelength, wave number, and name of radiation with an energy of 100 kJ/mol? (Answer: 1.20 m, 8.36 103 c m-1, infrared)
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Electromagnetic spectrum

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Electromagnetic spectrum

The diagram shows an approximation to the spectrum of visible light.

The main colour regions of the spectrum are approximately:

colour wavelength (nm) region violet blue cyan green yellow orange red 380 - 435 435 - 500 500 - 520 520 - 565 565 - 590 590 - 625 625 - 740

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Common Wavelength Symbols and Units

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interaction of light with matter;

Spectroscopy is the study of the interaction of radiant energy (light) with matter. Spectroscopy studies the interaction of light with matter defined as materials composed of molecules or atoms or ions. In a gas, atoms or molecules are widely separated from each other; in liquids and solids, the atoms or molecules are closely associated. In solids, the atoms or molecules may be arranged in a highly ordered array, called a crystal, as they are in many minerals, or they may be randomly arranged, or amorphous, as they are in many plastics. Whatever their physical state or arrangement atoms, molecules, and ions are in constant motion. For molecules, many types of motion are involved. Molecules can rotate, vibrate, and translate (move from place to place in space). Interaction with radiant energy can affect these molecular motions.

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Molecules that absorb IR radiation vibrate with greater amplitude; interaction with UV or visible light can move bonding electrons to higher energy levels in molecules. A change in any form of motion or electron energy level involves a change in the energy of the molecule. Such a change in energy is called a transition;

vibrational transitions, rotational transitions, electronic transitions, and so on in molecules.

We have some of the same kinds of motion in atoms and ions; atoms can move in space, and their electrons can move between energy levels, but atoms or monoatomic ions cannot rotate or vibrate. The chemical nature of matter (its composition), its physical state, and the arrangement of the atoms or molecules in the physical state with respect to each other affect the way in which any given material interacts with electromagnetic radiation.

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Some of the important types of transitions studied by spectroscopy

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When light strikes a sample of matter, the light may be

absorbed by the sample, transmitted through the sample, reflected off the surface of the sample, or scattered by the sample.

Samples can also emit light after absorbing incident light; such a process is called luminescence. There are different kinds of luminescence, called
fluorescence or phosphorescence depending on the specific process that occurs; For the moment, we will focus on the absorption, transmission, and emission of light by matter.

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Absorbance of light
When a molecule absorbs a photon, the energy of the molecule increases. We say that the molecule is promoted to an excited state. If a molecule emits a photon, the energy of the molecule is lowered. The lowest energy state of a molecule is called the ground state.

microwave radiation stimulates rotation of molecules when it is absorbed. Infrared radiation stimulates vibrations. Visible and ultraviolet radiation promote electrons to higher energy orbitals. X-rays and short-wavelength ultraviolet radiation break chemical bonds and ionize molecules.

Absorption of light increases the energy of a molecule. Emission of light decreases its energy

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When light is absorbed by a sample, the irradiance of the beam of light is decreased. Irradiance, P, is the energy per second per unit area of the light beam.

Schematic diagram of a single-beam spectrophotometric experiment. P0, irradiance of beam entering sample; P, irradiance of beam emerging from sample; b, length of path through sample

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Light is passed through a monochromator (a prism, a grating, or even a filter) to select one wavelength (Color Plate 12). Light with a very narrow range of wavelength is said to be monochromatic (one color.) The monochromatic light, with irradiance P0, strikes a sample of length b. The irradiance of the beam emerging from the other side of the sample is P. Some of the light may be absorbed by the sample,
so P P0

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Transmittance, T, is defined as the fraction of the original light that passes through the sample.
Transmittance: T= P/P0

Therefore, T has the range 0 to 1. The percent transmittance is simply 100T and ranges between 0 and 100%. Absorbance is defined as

Absorbance: A=log(P/P0) = -log T

When no light is absorbed, P =P0 and A= 0. If 90% of the light is absorbed, 10% is transmitted and P = P0/10. This ratio gives A=1. If only 1% of the light is transmitted, A =2. Absorbance is sometimes called optical density. Absorbance is directly proportional to the concentration, c, of the light-absorbing species in the sample
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Beers law:
Beers law:

A= bc Where, Absorbance: A, dimensionless Path lengh, b; expressed in centimeters molar absorptivity (or extinction coefficient, , M-1 cm-1 The heart of spectrophotometry as applied to analytical chemistry, is called the BeerLambert law, or simply Beers law. Molar absorptivity is the characteristic of a substance that tells how much light is absorbed at a particular wavelength.
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Absorbance, Transmittance, and Beers Law

Find the absorbance and transmittance of a 0.002 40 M solution of a substance with a molar absorptivity of 313 M1 cm-1in a cell with a 2.00-cm pathlength

Solution: Absorbance, A= bc =(313 M-1 cm-1)(2.00 cm)(0.002 40 M)= 1.50

Transmittance is obtained by raising 10 to the power equal to the expression on each side of the equation: T = 10 logT; T = 10A = 101.50 = 0.0316 Just 3.16% of the incident light emerges from this solution.

Try yourself: The transmittance of a 0.010 M solution of a compound in a 0.100-cmpathlength cell is T 8.23%. Find the absorbance (A) and the molar absorptivity (). (Answer: 1.08, 1.08 103 M-1 cm-1)

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Beers law:
A l = l bc

because A and depend on the wavelength of light. The quantity is simply a coefficient of proportionality between absorbance and the product bc. The greater the molar absorptivity, the greater the absorbance. The part of a molecule responsible for light absorption is called a chromophore. Any substance that absorbs visible light appears colored when white light is transmitted through it or reflected from it. (White light contains all the colors in the visible spectrum.) The substance absorbs certain wavelengths of the white light, and our eyes detect the wavelengths that are not absorbed. The observed color is called the complement of the absorbed color. For example, bromophenol blue has maximum absorbance at 591 nm and its observed color is blue.
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Colors of visible light

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Deviations from Beers Law /When Beers Law Fails

Beers law states that absorbance is proportional to the concentration of the absorbing species. It applies to monochromatic radiation, and it works very well for dilute solutions ( 0.01 M) of most substances. There are several possible reasons for deviation from linearity at high concentrations.

In concentrated solutions, solute molecules influence one another as a result of their proximity. When solute molecules get close to one another, their properties (including molar absorptivity) change somewhat. At very high concentration, the solute becomes the solvent. (6 Properties of a molecule are not exactly the same in different solvents. Non-absorbing solutes in a solution can also interact with the absorbing species and alter the absorptivity. If the absorbing molecule participates in a concentration-dependent chemical equilibrium, the absorptivity changes with concentration. For example, in concentrated solution, a weak acid, HA, may be mostly undissociated. As the solution is diluted, dissociation increases. If the absorptivity of A is not the same as that of HA, the solution will appear not to obey Beers law as it is diluted.

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Measuring Absorbance

The minimum requirements for a spectrophotometer (a device to measure absorbance of light) were shown in Fig

Figure outlines a single-beam instrument, which has only one beam of light Light from a continuous source is passed through a monochromator, which selects a narrow band of wavelengths from the incident beam. This monochromatic light travels through a sample of pathlength b, and the irradiance of the emergent light is measured.

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i.

ii. iii. iv. v.

vi.

We do not measure the incident irradiance, P0 , directly. Rather, the irradiance of light passing through a reference cuvet containing pure solvent (or a reagent blank) is defined as P0. This cuvet is then removed and replaced by an identical one containing sample. The irradiance of light striking the detector after passing through the sample is the quantity P. Knowing both P and P0 allows T or A to be determined. The reference cuvet compensates for reflection, scattering, and absorption by the cuvet and solvent. A double-beam instrument splits the light to pass alternately between sample and reference cuvets.

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For visible and ultraviolet spectroscopy, a liquid sample is usually contained in a cell called a cuvet that has flat, fused-silica (SiO2) faces. Glass is suitable for visible, but not ultraviolet, spectroscopy because it absorbs ultraviolet radiation. The most common cuvets have a 1.000-cm pathlength For infrared measurements, cells are commonly constructed of NaCl or KBr. For the 400 to 50 cm1 far-infrared region, polyethylene is a transparent window.

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Cuvets

Common cuvets for visible and ultraviolet spectroscopy. Flow cells permit continuous flow of solution through the cell. In the thermal cell, liquid from a constant-temperature bath flows through the cell jacket to maintain a desired temperature.
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Solid samples are commonly ground to a fine powder, which can be added to mineral oil (a viscous hydrocarbon also called Nujol) to give a dispersion that is called a mull and is pressed between two KBr plates. Solids and powders can also be examined by diffuse reflectance, in which reflected infrared radiation, instead of transmitted infrared radiation, is observed. Wavelengths absorbed by the sample are not reflected as well. Gases are more dilute than liquids and require cells with longer path lengths, typically ranging from 10 cm to many meters. A path length of many meters is obtained by reflecting light so that it traverses the sample many times before reaching the detector.

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In recording an absorbance spectrum, first record a baseline spectrum with reference solutions (pure solvent or a reagent blank) in both cuvets. If the instrument were perfect, the baseline would be 0 everywhere. In our imperfect world, the baseline usually exhibits small positive and negative absorbance. We subtract the baseline absorbance from the sample absorbance to obtain the true absorbance at each wavelength.

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For spectrophotometric analysis, we normally choose the wavelength of maximum absorbance for two reasons:
(1) The sensitivity of the analysis is greatest at maximum absorbance; that is, we get the maximum response for a given concentration of analyte. (2) The curve is relatively flat at the maximum, so there is little variation in absorbance if the monochromator drifts a little or if the width of the transmitted band changes slightly.

Beers law is obeyed when the absorbance is constant across the selected waveband.

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Beers law in Chemical Analysis

For a compound to be analyzed by spectrophotometry, it must absorb light, and this absorption should be distinguishable from that due to other substances in the sample. Because most compounds absorb ultraviolet radiation, measurements in this region of the spectrum tend to be inconclusive, and analysis is usually restricted to the visible spectrum. If there are no interfering species, however, ultraviolet absorbance is satisfactory. Proteins are normally assayed in the ultraviolet region at 280 nm because aromatic groups present in virtually every protein have an absorbance maximum at 280 nm.

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Example: Measuring Benzene in Hexane

A) Pure hexane has negligible ultraviolet absorbance above a wavelength of 200 nm. A solution prepared by dissolving 25.8 mg of benzene (C6H6, FM 78.11) in hexane and diluting to 250.0 mL had an absorption peak at 256 nm and an absorbance of 0.266 in a 1.000cm cell. Find the absorptivity of benzene at this wavelength. Solution: The concentration of benzene is [C6H6] = (0.025 8 g)/(78.11 g/mol)/0.250 0 L = 1.32x103 M We find the molar absorptivity from Beers law: Molar absorptivity =A /bc = (0.266)/(1.00 cm)(1.32x103 M) = 201.3 M-1 cm-1

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B) A sample of hexane contaminated with benzene had an absorbance of 0.070 at 256 nm in a cuvet with a 5.000-cm pathlength. Find the concentration of benzene in mg/L. Solution Using Beers law with the molar absorptivity from part (a), we find [C6H6] =A/b =0.070/2013 M1 cm1(5.00 cm) = 6.95 x105 M [C6H6] = (6.95x10-5 mol/L)(78.11 X103 mg/mol) = 5.4 mgL-1
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OPTICAL SYSTEMS USED IN SPECTROSCOPY

The instruments that are used to study the absorption or emission of electromagnetic radiation as a function of wavelength are called spectrometers or spectrophotometers. The essential components of a spectrophotometer include:

1- A stable source of radiant energy 2- A system of lenses, mirrors, and slits which define, collimate (make parallel) and focus the beam. 3- Monochromators to resolve the radiation into component wavelengths or bands of wavelength. 4- A transparent container to hold the sample. 5- Radiation detector 6- Readout system (meter, recorder or computer).
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The specific arrangement of these components is referred to as the optics or optical configuration or optical layout of the instrument. The optical layout of a simple single-beam absorption spectrometer is

:
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Radiation Sources
Sources of radiant energy consist of materials that are excited by a high voltage electric discharge or by electrical heating. As the materials return to lower energy states, they emit photons of characteristic energies corresponding to E, the energy difference between the excited and lower quantum states. An ideal radiation source for spectroscopy should have the following characteristics:

1. The source must emit radiation over the entire wavelength range to be studied. 2. The intensity of radiation over the entire wavelength range must be high enough so that extensive amplification of the signal from the detector can be avoided. 3. The intensity of the source should not vary significantly at different wavelengths. 4. The intensity of the source should not fluctuate over long time intervals. 5. The intensity of the source should not fluctuate over short time intervals. Short time fluctuation in source intensity is called flicker.

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Continuum sources
Continuum sources emit radiation over a wide range of wavelengths and the intensity of emission varies slowly as a function of wavelength. Typical continuum sources include:

the tungsten filament lamp which produces visible radiation (white light), the deuteriumlamp for the UV region, high pressure mercury or xenon arc lamps for the UV region, and heated solid ceramics or heated wires for the IR region of the spectrum. Xenon arc lamps are also used for the visible region.

Continuum sources are used for most molecular absorption and fluorescence spectrometric instruments.

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Line sources,
Line sources emit only a few discrete wavelengths of light, and the intensity is a strong function of the wavelength. Typical line sources include

hollow cathode lamps and electrodeless discharge lamps, used in the UV and visible regions for AAS and atomic fluorescence spectrometry, sodium or mercury vapor lamps (similar to the lamps now used in street lamps) for lines in the UV and visible regions, and lasers. Lasers are high intensity coherent line sources; lasers are available with emission lines in the UV, visible, and IR regions. They are used as sources in Raman spectroscopy, molecular and atomic fluorescence spectroscopy.
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Sources of Ultraviolet Radiation The hydrogen lamp and deuterium lamp are the most common sources of UV radiation. They consist of a pair of electrodes which are enclosed in a class tube provided with a quartz window and filled with hydrogen or deuterium gas at low pressure. When a stabilized high voltage is applied to the electrodes, an electron discharge occurs which excites other electrons in the gas molecules to high energy states. As the electrons return to their ground states they emit radiation in the region roughly between 180 and 350 nm. Sources of Visible Radiation A tungsten (W) filament lamp is the most satisfactory and inexpensive source of visible radiation. The filament is heated by a d-c power supply, or by a storage battery. The tungsten filament emits continuous radiation in the region between 350 and 2500 nm. Sources of Infrared Radiation The Globar and Ernst glower are the primary sources of infrared radiation. The Globar is a silicon carbide (SiC) rod heated to approximately 1200 0C. It emits continuous radiation in the (1-40) m region.

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Wavelength Selectors
These are devices which resolve wide band polychromatic radiation from the source into narrow bands or, even better, monochromatic radiation. There are two types of resolving devices

filters and mono-chromators. Filters prepared from special materials allow transmission of only limited wavelength regions while absorbing most of the radiation of other wavelengths. The simplest and most inexpensive way to select certain portions of the electromagnetic spectrum is with a filter. There are two major types:
absorption filters and interference filters.

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Absorption filters

Absorption filters can be as simple as a piece of colored glass. Colored glass absorption filters can be purchased that isolate various ranges of visible light. These filters are stable, simple, and cheap, so they are excellent for use in portable spectrometers designed to be carried into the field. The biggest limitation is that the range of wavelengths transmitted is broad compared with prisms and gratings which are also devices used to select a narrow wavelength range from a broad band polychromatic source. The transmission range may be 50300 nm for typical absorption filters. Absorption filters are limited to the visible region of the spectrum and the X-ray region.
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interference filter

the interference filter is constructed of multiple layers of different materials. The filter operates on the principle of constructive interference to transmit selected wavelength ranges. The wavelengths transmitted are controlled by the thickness and refractive index of the center layer of material. Interference filters can be constructed for transmission of light in the IR, visible, and UV regions of the spectrum. The wavelength ranges transmitted are generally 110 nm, The amount of light transmitted is generally higher than for absorption filters.

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Monochromators

Monochromators resolve poly-chromatic into its individual wavelengths and isolate these wavelengths into very narrow bands. The components of a monochromator include:
(1) an entrance slit which admits polychromatic radiation from the source; (2) a collimating device either a lens or a mirror; (3) a dispersing device, either a prism or grating which resolve the radiation into small bands of wavelengths emerging at different angles; (4) A focusing lens or mirror; (5) An exit slit.
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Sample Containers

The cells or cuvettes that hold the samples must be made of material that is transparent to radiation in the spectral region of interest. Quartz or fused silica is required for work in the ultraviolet region (< 350 nm). Plastic containers have also found application in the visible region. Crystalline sodium chloride (NaCl) is the most common substance employed for cell windows in the infrared region.
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Radiation Detectors
Any detector absorbs the energy of the radiation and converts this energy to a measurable quantity such as the darkening of a photographic plate, electric current or thermal changes. Any detector must generate a signal which is quantitatively related to the radiant power striking it. The noise of a detector refers to the background signal or dark current generated when no radiant power from the sample reaches the detector.

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Phototubes Ultraviolet and visible radiation, possess enough energy to cause photoejection of electrons when they strike surfaces which have been treated with specific type of compounds. Their absorption may also cause bound, nonconducting electrons to move into conducting bands in certain semiconductors. Both processes generate an electric current which is directly proportional to the radiant power of the absorbed radiation. Photomultiplier tube If the ejected electron is accelerated by an electric field, it acquires more energy; and if it strikes another electron-active surface, it may transfer some of its energy, ejecting several more electrons. These electrons may in turn be accelerated to another surface and produce even more electrons, and so on. Each succeeding electron active plate, or dynode, is at higher electrical potential and thus acts as an amplification stage for the original photon. After nine stages of amplification, the original photon has been amplified by a factor of approximately 106.
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Photoconductivity detectors These are semiconductors whose resistance decreases when they absorb radiation in the region (0.75 to 3 m). The application of photoconductors is important in Fourier Transform Infrared instrumentation FTIR. Absorption of radiation by semiconductor materials promotes some of their bound electrons into an energy state in which they are free to conduct electricity. The resulting change in conductivity can then be measured. Thermal detectors In thermal detectors, the radiation absorbed is converted to thermal energy (heat) and a corresponding temperature change is noted. These are various types of rapid response thermometers such as thermocouples, resistance thermometers (bolometer), gas thermometers, and pyroelectric transducers.

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Processors and readout

The electronic signal generated by any radiation detector must be translated into a form that can be interpreted. This process is typically accomplished with amplifiers, ammeters, potentiometers and potentiometer recorders. Amplifiers

The amplifier takes an input signal from the circuit of the sensing component and, through a series of electronic operations, produces an output signal which in many times larger than the input. The amplification factor (ratio of output to input) is called the gain of the amplifier.

Readout devices
Several types of readout devices are found. For example, the digital meters, the scale of potentiometers, cathode ray tubes and computers. The instrument is calibrated so that there are 100 units on the meter from (It=0) to (I = I0) and these units are linear with respect to It. When an absorbing sample is substituted for the blank, the detector response will show between 0 and 100 units on the meter

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