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1. 2. Introduction of spectroscopy UV-Visible spectroscopy (Electronic Spectroscopy) 3. Infrared spectroscopy ( IR spectroscopy) 4. Microwave spectroscopy Studying the properties of matter through its interaction with different frequency components of the electromagnetic spectrum. The term "spectroscopy" defines a large number of techniques that use radiation to obtain information on the structure and properties of matter. The basic principle shared by all spectroscopic techniques is to pass a beam of electromagnetic radiation onto a sample, and observe how it responds to such radiation. The response is usually recorded as a function of radiation wavelength. A plot of the response as a function of wavelength is referred to as a spectrum. It is the most powerful tool available for the study of atomic and molecular structures. It is used in the analysis of wide range of samples. Electromagnetic radiation is an oscillating electric and magnetic disturbance that spreads as a wave through empty space, the vacuum. electromagnetic radiation consists of electromagnetic waves, which are synchronized oscillations of both electric and magnetic fields which are perpendicular to each other. Electromagnetic waves are transverse in nature as they propagate by varying the electric and magnetic fields such that the two fields are perpendicular to each other. Accelerated charges are responsible to produce electromagnetic waves. The energy changes within a molecule during emission or absorption electromagnetic radiation are quantized. Each particular frequency of light has a particular energy associated with it, given by another simple equation. E=hv E=energy of radiation h=planck’s Constant v=Frequency of radiation TYPE 1: A) Atomic Spectroscopy : It is concerned with interactions of Electro Magnetic Radiations with atoms. B) Molecular Spectroscopy : It is concerned with interactions of electro Magnetic Radiations with molecules TYPE 2: A) ABSORPTION SPECTRUM: If the electromagnetic radiations are passed through a substance, the dark pattern of lines that are obtained corresponding to wave lengths absorbed is known as Absorption spectrum. B) EMISSION SPECTRUM: If electromagnetic radiations are passed through a substance, the pattern of lines recorded after the emission of the absorbed wave lengths are known as emission spectrum. The theory revolving around this concept states that the energy from the absorbed ultraviolet radiation is actually equal to the energy difference between the higher energy state and the ground state. UV spectrophotometer principle follows the Beer-Lambert Law. This law states that whenever a beam of monochromatic light is passed through a solution with an absorbing substance, the decreasing rate of the radiation intensity along with the thickness of the absorbing solution is actually proportional to the concentration of the solution and the incident radiation. This law is expressed through this equation: A = log (I0/I) = ECI A stands for the absorbance, I0 refers to the intensity of light upon a sample cell, l refers to the intensity of light departing the sample cell, C stands for the concentration of the solute, L stands for the length of the sample cell and E refers to the molar absorptivity. The absorption of UV or visible radiation corresponds to the excitation of outer electrons. Transitions are possible by involving π,σ, and n electrons. When an atom or molecule absorbs energy, electrons are promoted from their ground state to an excited state. In a molecule, the atoms can rotate and vibrate with respect to each other. These vibrations and rotations also have discrete energy levels, which can be considered as being packed on top of each electronic level. The ultraviolet region falls in the range between 200-400 nm, visible region fall between 400-750 nm. Absorption of radiation in this region causes electronic transitions between electronic levels. This absorption changes vibrational and rotational levels also. σ–σ* Transitions An electron in a bonding s orbital is excited to the corresponding antibonding orbital. The energy required is large. For example, methane (which has only C-H bonds, and can only undergo σ–σ* transitions) shows an absorbance maximum at 125 nm. Absorption maxima due to σ– σ* transitions are not seen in typical UV-Vis. spectra (200 - 700 nm) n–σ* Transitions Saturated compounds containing atoms with lone pairs (non-bonding electrons) are capable of n–σ* transitions. These transitions usually need less energy than σ–σ* transitions. They can be initiated by light whose wavelength is in the range 150 - 250 nm. The number of organic functional groups with n–σ* peaks in the UV region is small. n–π* and π–π* Transitions Most absorption spectroscopy of organic compounds is based on transitions of n or π electrons to the π excited state. This is because the absorption peaks for these transitions fall in an experimentally convenient region of the spectrum (200 - 700 nm). These transitions need an unsaturated group in the molecule to provide the p electrons. Molar absorbtivities from n–π* transitions are relatively low, and range from 10 to100 L mol-1 cm-1 . π–π* transitions normally give molar absorbtivities between 1000 and 10,000 L mol-1 cm-1 . When ultraviolet radiations are absorbed, this results in the excitation of the electrons from the ground state towards a higher energy state. Molecules containing π-electrons or non-bonding electrons (n-electrons) can absorb energy in the form of ultraviolet light to excite these electrons to higher anti-bonding molecular orbitals. The more easily excited the electrons, the longer the wavelength of light it can absorb. There are four possible types of transitions (π–π*, n–π*, σ– σ*, and n–σ*), and they can be ordered as follows: σ–σ* > n–σ* > π–π* > n–π* The absorption of ultraviolet light by a chemical compound will produce a distinct spectrum which aids in the identification of the compound. Detection of Impurities It is one of the best methods for determination of impurities in organic molecules. Additional peaks can be observed due to impurities in the sample and it can be compared with that of standard raw material. By also measuring the absorbance at specific wavelength, the impurities can be detected. Structure elucidation of organic compounds It is useful in the structure elucidation of organic molecules, such as in detecting the presence or absence of unsaturation, the presence of hetero atoms. UV absorption spectroscopy can be used for the quantitative determination of compounds that absorb UV radiation. UV absorption spectroscopy can characterize those types of compounds which absorbs UV radiation thus used in qualitative determination of compounds. Identification is done by comparing the absorption spectrum with the spectra of known compounds. This technique is used to detect the presence or absence of functional group in the compound. Absence of a band at particular wavelength regarded as an evidence for absence of particular group. Kinetics of reaction can also be studied using UV spectroscopy. The UV radiation is passed through the reaction cell and the absorbance changes can be observed. Many drugs are either in the form of raw material or in the form of formulation. They can be assayed by making a suitable solution of the drug in a solvent and measuring the absorbance at specific wavelength. Molecular weights of compounds can be measured spectrophotometrically by preparing the suitable derivatives of these compounds. UV spectrophotometer may be used as a detector for HPLC. MICROWAVE-SPECTROSCOPY APPLICATIONS