— CHM 2 bee. cons . RIC EAE YO eng COk Sey og|202 OPTICAL METHODS OF ANALYSIS tal id Interaction of radiation ap matter i radiation isa form of enedgy which could ether bp absorbed or transmitted, The interaction of radiation in ‘avolves transition between the different energy levels of atoms ot molecules. The other types of spectroscopy interactions such as reflection, refraction and diffraction, are often related to the bulky properties of materials rather than to energy levels'of specific atoms or molecules. Generally, the absorption or emission of radiation IHange of energy. Thus, to understand the principle of this exchange, itis necessary by matter involves the to know the distribution of energy within an atom or molecule, The internal energy of a molecule is due to the atoms (iii). the rotation of various grou energy associated with (i), the electrons (ji) vibrations betw atoms within a molecule. 1 be altered by the absoiption dr emission of radiant energy. This is because atoms exist within a limited number of energy levels. A sthdy of the wavelength or frequency of radiation absorbed Jecule will ‘give information about its identity. This technique is known as or emitted by an atom qualitative spectroscop y Electromagnetic radiation ectromagnel fonn of energy which has both magnetic and electrical properties, hence called Radiatio radiation. The electromagnetic ‘radiation covered a long range of radiations which ere described o: haracterized by either wavelength or frequency. e peaks which is nieasured Wavelength (is defined as the distance betwe (am). the number of successive peaks passing a given poi squency (of radiation is defined relationship between the ovo parameters is: 1 proportional to the frequency and inversely proportional 10 wavelenWavelength (m), a 10° | 10° 10 2 i | | las 2 ° Infrared x 3 Visible Microwaves Radio waves Ultraviolet Wavelength (nm) | Fig. 11: Range of eleetromagnetic radiation Absorption of radiation Every molecular specie is capable of absorbing: s own characteristic frequencies or wavelength of electromagnetic radiation. ‘This process transfers enbrgy to the molecule or atom and results in a decrease in the intensity of the incidentiradiation, Thus, absorption of radiation could be: (). Atomic absorption which involves the absorption of electromagnetic radiation by free atomic species. The to the lowest excited state Known as the resonance line wavelength that is most = absorbed usually edresponds toan electronic transition from the ground state (i), Molecular absorption which involves the absozption of radiation By molecules in solution. This type of absorption usually takes pldce in ultraviolet and visible region of the electromagnetic spectrum. Absorption of radiation in this region cases transition of electro from molecular bonding orbital to the higher energy i molecular antibonding orbital Emission of radiation When an atom or molecule absorbs electromagnetic radiation, it will result in ether electonie or vibrational transition from lower to higher energy level. Such atom or molecule is said tobe in an excited or unstable state. If the atom or molecule loses all or part of this enetgy as radiation, photons of energy will be emitted which ] levels involved. The radiation emitted is of specific correspond to the differerjce between the energy frequencies and will show gp as a bright line lait as a spectrum. Just like absorption, emission could | also be atomic or moleculat. | ' 244 by many elements; particularly the metals which after being excited either Atomic emission is displa thermally ot electrically emt discontinuous lines spectrum ove to transition ending in the ground state Molecular emissions are due to electronic transition within the molecule but are modified or influenced by of sions are more complex than atomic emissions. The radiation emitted con: bond length. Molecular emis ian the narrow lines associated with atomic emission, broad bands of radiation rpther th also the resulting spectrum is approximately fhe mirror image of the absorption spectrum of the compound SPECTROCHEMICAL OPTICAL M! THODS Introduction | atensity. To relate optical measurement 10 al method of analysis involves the measurement of lis Opti amount of a particular substance in a sample, 18 the ability of atoms sigular set of conditions. Thus, this method of analysis absorb of emit light radiation when exposed fo # Pp cation, avolves the use of instrument with varying degrees of sophi Definition: Optical methogs are spectroscopic techniques that are based on the absorption oF emiss frared radiation. ‘Types of optical methods of analysis ultraviolet, visible and i soluti nent ofthe absorption of light radiation by # colo {i). Colorimetry which involves the measur 1 narrow wavelength band by « sures U Spectrophotometry which solution, ation light radiation by a free atomic specie, i), Atomic absorption analysis which measures the 2bs Auoregeent substance ina given samp of the amor (iii). Flourimetry which inyolves the estimat nission light atoms in exited electronic state 5, Anvil \ jv), Atomic emission analysis which based on the © rent of the absorption of visible radiation of light BY @ c jorimetry this method of analysis involves the measuren nent is carried out by the use of an instrament called colorimeter. The olution. This type of me colorimeter consists of:ight souree (tungsten Lani) Monochromator (Filter), Slit, Optical cell or vette anc Photo electric Light source (tungste . cell Galvanometer i i ‘The instrumental set up is t bellow: j=l Light source Monochromator — @-@ Detector Optical cell Schematic diagram of a simple Colorimeter Mode of operation | White light from a singin tamp passes through a bondenser lens to give a parallel bear which falls on the filter that is positioned to select radiation of specifid wavelength to impinge on a glass cuvete containing the s the light is pagsing through the solution, some part of itis absorbed by the sample component, cell (detector). In order to t absorbed is transmitted, apd detected by a photo electric cel Jution, the meter reading is first adjusted to 100% es then inserted in place ofthe blank andthe absorbance vead solution. while the part that is not transmittance (zero measure the absorbance of a sol n. The samp! gto the absorbance of the sample is then obtained from the standard absorbance) with a blank solu directly. The concentration correspon: or calibration graph. Spectrophotometry | | | ‘This method of analysis involves the measurement of the absorption ofa narcow wavelength band of radiation olution, The instruments that are Used to study or to measure the absorption or er wave lergth are called spectrometers or spectrophatometers, T by molecules in s mission of electromagnetic radiation 4 a function of | optical and electronic principles employed in these instruments are basically the same forall the rewions of Fthe spectrum, The essential components of a speotrophotometer include: stable source of radiant energy. ‘ors and slits which define collimate (malee parallel) and focus the beam of light. 4 system of lenses, mirr ! 4monochromator to resolve the radiation into its individual wavelength cuvette or cell for holding the sample radiation detector with an associated read out system (meter or recorder) Below is a schematic diagram of a typical spectrophotometer: rx B44 2] jetector Meter Radiation souree Monechromator cell Sample Components of a Spectrophotometer Radiation source ps consist of a « soutce of ultraviolet radiation is provided by hydrogen and deuterium lamps, These ja Jass tube provided with a quartz window and f of 180 ~350nm ed with hydr pait of electrodes whici aré enctosed in a gl cor deuterium gas at low pressure, These lamps provide radiation in the ran lament lamp is the most satisfactory Source of visible and near infrared radia A tungster jon in th provides radiati he région between 350 and 2500nm, Other sources of radiation include: xenon lamp. Globar and Nemst glower Monochromator 0 its individual wavelength somator is a device which is used to resolve polychromatic radiation Monod radiation of only particular and isolates these wavelengitis into a very narrow band. This device produces ligh’ led monvchromatic radiation 4. ath isc (single) wavelength. Thus, light of single or only one, wavelet Jiation ftom the source: a monochromator consists of an entrance slit which admits polychromatic ich resolves collimating device which is either a lens or a mirror; a dispersion device, prism or g radiation into its individual components, a focusing lens or mitror; and an exit slitnd translates the transmitted radiation from the sample into an electrical photographie film can be used as a detector. A detector 1, It is capable of absorbing the energy of photons the detection of ultraviolet and Detectors ‘Adetector isa device which identifies a Signal that activates a mete or recorder. Example, + dicates the existence of some physical phebomenot measfrable quantity. Photoeletic detectors are used for classified as phototubes and photoelectric cells. usually in and converts it into a visible radiations. These detectors are y used for the detection of near infrared radiation, ‘Another detector is photocdnduetive cell which is eemmon! But for middle and far infrared radiation, thermocouple detectors are used. erally, the detectors us@d in spectrophotometrie jnstruments could be: + photoelectric or barrier cel * photomultiplier tube. + thermocouple. + photoconductivity cell Differences between colorimeter and spectrophofometer | Unlike colorimeter, spectrdphotometer can discriminate effectively between compounds with overlapping absorption spectra, The filter system found in colorirpeter is replaced by prism or grating in spectrophotometer ‘The light passing through the sample solution is detgcted by a photocell in colorimeter, while itis detected by ne photomultiplier tube in most spectrophotometers. With most spectrophotometers, measurements can be do} n both ultraviolet and visfble region of the specttum, which is not possible with eolorimeters. Thus, the advantages displayed by spectrophotometer in comparison with colorimeter are improved resolution and sensitivity together with greater versatility. The more elaborate spectrophotometers have a scenning device which allows for the automatic determination of the absorbance of a solution as a continuous function of wavelength. Spectrophotometers with direct reading device are also available.Flame Spectroscopy Introduetiot Ifa solution containing a metallic salt, e.g, sodium chloride is aspirated into a flame for example acetylene burning in sir, @ vapour which contains atoms of the metal may be formed. Some of these gaseous metal atoms may be raised to an energy level which is sufficiently high to permit the emission of radiation characteristics|of the metal, e.g. the cljarncteristies yellow colour imparted to Names by compounds of sodium. Thip is the basis of fame photometry. However, a much larger number of the gaseous metal atoms will mrmally cemain in an unexcited state or, in other words, in the ground stare These grounds-state atoms| are capable of absorbing radiant energy of their own specific resonance wavelength, which in genefal isthe wavelength of the radiation that the toms would emit if excited from the ground state. Hence, iflight of the resonante wavelength is passed through a flame containing of the light will be absorbed, and the extent of absorption will be the atoms in question, then part s is the underlying principle proportional to the number of ground state atoms present in the fame, This of atomic absorption spectroscopy (AAS). | | Definition of Flame Spectroscopy | « Flame Emission Spectrosdopy isa technique ia whjeh the emission of light by thermally excited atoms in a flame or furnace is used to measure the concgntration of atoms, and Flame Atomic Absorption rptior| of light by free gaseous atoms in flame or furn Spectroscopy is a technique in which the abso is used to measure the concentration of atoms. Differences between Flame Emission and Flame Atomic Absorption Spectroscopy. Flame emission spectroscdpy is basically the sanje as flame atomic absorption spectroscopy. The difference is that no light source is needed in flame emission. Some of the atoms in the fame are collision|with other atoms. The e ited atoms entit their nic states to their ground tate. In flame emission spectroscopy promoted to excited ele the emission ( characteristic radiation as tey retum. intensity at a characteristic wavelength of an element is nearly proportional to the c element in the sample, For both absorption and emission, standard waves are usually used co establish the relation between signal and concentration.trometry (FES) fine spray or aerosol (Nebulization). The solvent issociated into free gaseous atoms in the ground rinciple of Flame Emission Spe Working Pt me as a lution is introduced into the fla /drated salt. The salt is can absorb nergy from the flame and be raised to an excited The sol porates leaving the deh} A certain fraction ct atoms eval state. clectronic state. The excited levels have a short liftime and! drop back to the ground state, emitting photons of characteristic swfelenath ‘These can be detected with conventional monochromator-detector get up. The intensity of emission is directly proportional to the concentration ofthe analyte in solution fc diagram of a flame emission spectrometer is given below: (EH=}0 being aspirated. A schema Monochromator ofFiter | | Meter Burner | .¢ emission spectrometer | | | | Working Principle of Flame Atomic Absorption Speetrophotometry (FAAS) ple solution is aspitated into a flame as in flame emission spectrometry and the sample The flame then coptains atoms of that element, Some are thermally A schematic diagram of a fam The sam element is converted to atomic vapour. excited by the lame, but must remain in the ground state, These ground state atoms ean absorb radiation ofa particular wavelength that is produced by a special source made from that element. The wavelength of radiation given off by the sources is the same as those absorbed by the atoms in the flame, The absorption follows Beer’s Law, that is, the absorbance is directly proportional to the path length in the fMame and to the concentration of atomic vapour il the flame, A schematic diagram of flame atomic absorption speetrophotometer is given below:| lamp Source [vetow \ Chopper Flame _ Cathode + roacrons peter |__| meter Ls | (dc supply) A schematic diagram of flame atomic absorption spectrophotometer Interference: By interference, we mean) any effect that changes the signal when analyte concentration remains jespreal unchanged. In the measurement of atomic absorptipn or emission signals, interference is wid and easy to overlook. If t are clever enough td'discern that interference is occurring, it may be corrected by counteracting {he source of interferencs Types of interference: Spectral: zefers to the overlap of analyte signal with signals due to other elements or molecules 1p Je or with signals due to flame or furnace. The best means of dealing with overlap betw the samp! lines of different elements in the sample is to cioose another wavelength for analysis. nt of atomization of used by any component of the sample that decreases the exte 2. Chemical: der the atomization of Ca®*, perhaps by forming non volatile salts. i analyte. E.g. SO." and POs “This can be solved by allding releasing agents which are chemicals to a sample to decrease chentisal terference, eg EDTA /and 8 ~ hydroxyquinolige protect Ca Tonization: In this, the ionization of analyte atpms decreases the concentration of n from the above interference. ral analyte ‘atoms in the flame, which results in the decrease of the desired atomic signal the extent of ionization This is solved by adding an ionization suppressor to'a sample so as to decreas of analyte. 9