Spectroscopy is the study of the interaction between electromagnetic radiation and matter. It involves using different types of radiation such as ultraviolet, visible, infrared to cause transitions between energy levels in atoms and molecules. This interaction provides information about the composition and structure of matter. Common spectroscopy techniques include UV-Vis, IR, NMR, and atomic spectroscopy which use electromagnetic radiation to determine elemental composition. Spectrometers measure the absorption, emission or fluorescence of samples to obtain spectra unique to different materials.
Spectroscopy is the study of the interaction between electromagnetic radiation and matter. It involves using different types of radiation such as ultraviolet, visible, infrared to cause transitions between energy levels in atoms and molecules. This interaction provides information about the composition and structure of matter. Common spectroscopy techniques include UV-Vis, IR, NMR, and atomic spectroscopy which use electromagnetic radiation to determine elemental composition. Spectrometers measure the absorption, emission or fluorescence of samples to obtain spectra unique to different materials.
Spectroscopy is the study of the interaction between electromagnetic radiation and matter. It involves using different types of radiation such as ultraviolet, visible, infrared to cause transitions between energy levels in atoms and molecules. This interaction provides information about the composition and structure of matter. Common spectroscopy techniques include UV-Vis, IR, NMR, and atomic spectroscopy which use electromagnetic radiation to determine elemental composition. Spectrometers measure the absorption, emission or fluorescence of samples to obtain spectra unique to different materials.
electromagnetic radiation. the interaction of ultraviolet, visible, and infrared radiation with matter. Because these techniques use optical materials to disperse and focus the radiation, they often are identified as optical spectroscopies. Electromagnetic Radiation
is a 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. 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. The oscillations in the electric and magnetic fields are perpendicular to each other, and to the direction of the waves propagation. an example of plane- polarized electromagnetic radiation, consisting of a single oscillating electric field and a single oscillating magnetic field.
Plane-polarized electromagnetic radiation showing the oscillating electric field in red
and the oscillating magnetic field in blue. The radiations amplitude, A, and its wavelength, , are shown. Normally, electromagnetic radiation is unpolarized, with oscillating electric and magnetic fields present in all possible planes perpendicular to the direction of propagation. INTERACTION OF ELECTROMAGNETIC RADIATION WITH MATTER The interaction of radiation with matter can cause redirection of the radiation and/or transitions between the energy levels of the atoms or molecules. 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. Photons is absorbed by a sample it is destroyed, and its energy acquired by the sample.3 The energy of a photon, in joules, is related to its frequency, wavelength, and wavenumber by the following equalities :
E = hv = = hc
where h is Plancks constant, which has a value of 6.626 1034 J .s. Common types of Spectroscopy X-ray X-rays of sufficient energy are used to excite the inner shell electrons in the atoms of a sample. The electrons move to outer orbitals then down into the vacated inner shells and the energy in this de-excitation process is emitted as radiation. The absorption or emission energies are characteristic of the specific atom and small energy variations may occur that are characteristic of particular chemical bonding. The X-ray frequencies can be measured and X-ray absorption and emission spectroscopy is used to determine elemental composition and chemical bonding. Common types of Spectroscopy Flame Usually the analyte is in solution form (or converted into one) that is then converted to a free gaseous form in a multistage process (atomization). This method is often used for metallic element analytes present at very low concentration ranges. Common types of Spectroscopy Visible/Ultraviolet (UV) This uses the fact that many atoms are able to emit or absorb visible light. The atoms must be in a gaseous phase in order to obtain a spectrum just as those obtained in flame spectroscopy. It is common for visible absorption spectroscopy to be combined with UV absorption spectroscopy in UV/Vis spectroscopy. UV spectroscopy can also be used to analyze fluorescence from a sample in a form of absorption spectroscopy. Common types of Spectroscopy Infrared (IR) and Near Infrared (NIR) IR spectroscopy is used to show what types of bonds are present in a sample by measuring different types of inter-atomic bond vibrations at different frequencies. It relies on the fact that molecules absorb specific frequencies which is dependent on their chemical structure. This is determined by factors such as the masses of the atoms. NIR shows a greater penetration depth into a sample than mid- infrared radiation. This indicates a low sensitivity but also that it allows large samples to be measured in each scan by NIR spectroscopy with little (if any) sample preparation. It has numerous practical applications that include: medical diagnosis pharmaceuticals, biotechnology, various analyses (genomics, proteomic) and chemical imaging of intact organisms, textiles, forensic lab application and various military applications. Common types of Spectroscopy Nuclear Magnetic Resonance This is a prominent method for analyzing organic compounds because it exploits the magnetic properties of certain atomic nuclei to determine the properties (both chemical and physical) of these atoms or the molecules containing them. It can provide extensive information about the structure, dynamics, and chemical environment of atoms. Additionally, even different functional groups are distinguishable, and identical functional groups in differing molecular environments still give distinguishable signals. Spectroscopic Analysis Atomic Spectroscopy Molecular Spectroscopy Atomic Spectroscopy the determination of elemental composition by its electromagnetic or mass spectrum. Electrons exist in energy levels within an atom. These levels have well defined energies and electrons moving between them must absorb or emit energy equal to the difference between them. The science of atomic spectroscopy has yielded three techniques for analytical use: Atomic Absorption, Atomic Emission, Atomic Fluorescence. The process of excitation and decay to the ground state is involved in all three fields of atomic spectroscopy. Either the energy absorbed in the excitation process, or the energy emitted in the decay process is measured and used for analytical purposes. Atomic Spectroscopy Atomic Absorption measures the amount of light at the resonant wavelength which is absorbed as it passes through a cloud of atoms. As the number of atoms in the light path increases, the amount of light absorbed increases in a predictable way. By measuring the amount of light absorbed, a quantitative determination of the amount of analyte element present can be made. atomic absorption is the most widely applied of the three techniques and usually offers several advantages over the other two, particular benefits may be gained with either emission or fluorescence in special analytical situations. Atomic Spectroscopy Atomic Emission a sample is subjected to a high energy, thermal environment in order to produce excited state atoms, capable of emitting light. The energy source can be an electrical arc, a flame, or more recently, a plasma. The emission spectrum of an element exposed to such an energy source consists of a collection of the allowable emission wavelengths, commonly called emission lines, because of the discrete nature of the emitted wavelengths. Atomic Spectroscopy Atomic Fluorescence This technique incorporates aspects of both atomic absorption and atomic emission. The intensity of this "fluorescence" increases with increasing atom concentration, providing the basis for quantitative determination. Molecular Spectroscopy the study of absorption of light by molecules. a given molecule will specifically absorb only those wavelengths which have energies that correspond to the energy difference of the transition that is occurring. In the gas phase at low pressures, molecules exhibit absorption in narrow lines which are very characteristic of the molecule as well as the temperature and pressure of its environment. In the microwave and long-wavelength infrared regions of the spectrum, these lines are due to quantized rotational motion of the molecule. At shorter wavelengths similar lines are due to quantized vibration and electronic motion as well as rotational motion. Instruments for measuring Absorption
INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION
INSTRUMENTS USED FOR INFRARED ABSORPTION INSTRUMENTAL COMPONENTS OF UV/VIS ABSORPTION Sources of UV radiation It is important that the power of the radiation source does not change abruptly over it's wavelength range. The electrical excitation of deuterium or hydrogen at low pressure produces a continuous UV spectrum. The mechanism for this involves formation of an excited molecular species, which breaks up to give two atomic species and an ultraviolet photon. INSTRUMENTAL COMPONENTS OF UV/VIS ABSORPTION Sources of visible radiation The tungsten filament lamp is commonly employed as a source of visible light. This type of lamp is used in the wavelength range of 350 - 2500 nm. The energy emitted by a tungsten filament lamp is proportional to the fourth power of the operating voltage. This means that for the energy output to be stable, the voltage to the lamp must be very stable indeed. Electronic voltage regulators or constant-voltage transformers are used to ensure this stability. INSTRUMENTAL COMPONENTS OF UV/VIS ABSORPTION Wavelength selector (monochromator) All monochromators contain the following component parts; An entrance slit A collimating lens A dispersing device (usually a prism or a grating) A focusing lens An exit slit Polychromatic radiation (radiation of more than one wavelength) enters the monochromator through the entrance slit. The beam is collimated, and then strikes the dispersing element at an angle. The beam is split into its component wavelengths by the grating or prism. By moving the dispersing element or the exit slit, radiation of only a particular wavelength leaves the monochromator through the exit slit. INSTRUMENTAL COMPONENTS OF UV/VIS ABSORPTION Cuvettes The containers for the sample and reference solution must be transparent to the radiation which will pass through them. Quartz or fused silica cuvettes are required for spectroscopy in the UV region. These cells are also transparent in the visible region. Silicate glasses can be used for the manufacture of cuvettes for use between 350 and 2000 nm. INSTRUMENTAL COMPONENTS OF UV/VIS ABSORPTION Detectors photomultiplier tube is a commonly used detector in UV-Vis spectroscopy. It consists of a photoemissive cathode (a cathode which emits electrons when struck by photons of radiation), several dynodes (which emit several electrons for each electron striking them) and an anode. linear photodiode array is an example of a multichannel photon detector. These detectors are capable of measuring all elements of a beam of dispersed radiation simultaneously. A linear photodiode array comprises many small silicon photodiodes formed on a single silicon chip. INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION Filter Photometer The simplest instrument for molecular UV/Vis absorption is a filter photometer which uses an absorption or interference filter to isolate a band of radiation. The filter is placed between the source and the sample to prevent the sample from decomposing when exposed to higher energy radiation. A filter photometer has a single optical path between the source and detector, and is called a single-beam instrument. The instrument is calibrated to 0% while using a shutter to block the source radiation from the detector. INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION Single-beam Spetrophotometer An instrument that uses a monochromator for wavelength selection The simplest spectrophotometer is a single-beam instrument equipped with a fixed-wavelength monochromator Single-beam spectrophotometers are calibrated and used in the same manner as a photometer. The accuracy of a single-beam spectrophotometer is limited by the stability of its source and detector over time. INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION Double-Beam Spectrophotometer The limitations of fixed-wavelength, single-beam spectrophotometers are minimized by using a double-beam spectrophotometer. A scanning monochromator allows for the automated recording of spectra. Double-beam instruments are more versatile than single-beam instruments, being useful for both quantitative and qualitative analyses, but also are more expensive. INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION Diode Array Spectrometer An instrument with a single detector can monitor only one wavelength at a time. If we replace a single photomultiplier with many photodiodes, we can use the resulting array of detectors to record an entire spectrum simultaneously in as little as 0.1 s. In a diode array spectrometer the source radiation passes through the sample and is dispersed by a grating. One advantage of a diode array spectrometer is the speed of data acquisition, which allows to collect several spectra for a single sample. One disadvantage of a photodiode array is that the effective bandwidth per diode is roughly an order of magnitude larger than that for a high quality monochromator. INSTRUMENTS USED FOR MOLECULAR UV/VIS ABSORPTION Sample Cells The sample compartment provides a light-tight environment that limits the addition of stray radiation. Samples are normally in the liquid or solution state, and are placed in cells constructed with UV/Vis transparent materials, such as quartz, glass, and plastic. INSTRUMENTS USED FOR INFRARED ABSORPTION Filter Photometer The simplest instrument for IR absorption spectroscopy is a filter photometer similar for UV/Vis absorption. These instruments have the advantage of portability, and typically are used as dedicated analyzers for gases such as HCN and CO. INSTRUMENTS USED FOR INFRARED ABSORPTION Double-beam spectrophotometer Infrared instruments using a monochromator for wavelength selection use double-beam optics. Double-beam optics are preferred over single-beam optics because the sources and detectors for infrared radiation are less stable than those for UV/Vis radiation. In addition, it is easier to correct for the absorption of infrared radiation by atmospheric CO2 and H2O vapor when using double-beam optics INSTRUMENTS USED FOR INFRARED ABSORPTION Fourier transform spectrometer In a Fourier transform infrared spectrometer, or FTIR, the monochromator is replaced with an interferometer. Because an FT-IR includes only a single optical path, it is necessary to collect a separate spectrum to compensate for the absorbance of atmospheric CO2 and H2O vapor. This is done by collecting a background spectrum without the sample and storing the result in the instruments computer memory. The background spectrum is removed from the samples spectrum by ratioing the two signals. In comparison to other instrument designs, an FTIR provides for rapid data acquisition, allowing an enhancement in signal-to-noise ratio through signal-averaging. INSTRUMENTS USED FOR INFRARED ABSORPTION Sample Cells Infrared spectroscopy is routinely used to analyze gas, liquid, and solid samples. Sample cells are made from materials, such as NaCl and KBr, that are transparent to infrared radiation. Gases are analyzed using a cell with a pathlength of approximately 10 cm. Longer pathlengths are obtained by using mirrors to pass the beam of radiation through the sample several times. Volatile liquids must be placed in a sealed cell to prevent their evaporation.