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TABLES OF CONTENTS
NO 1.0 UV-VIS SPECTROSCOPY 2.1 Introduction 2.2 2.3 2.4 2.5 2.0 Theory and principles Absorption process (beer-lambert law) Instrument of uv vis spectroscopy Instrument schematic diagram TOPICS PAGES

FOURIER-TRANSFORM SPECTROSCOPY 2.1 Introduction 2.2 History 2.3 Theory and principles 2.4 Instrument schematic diagram

3.0

REFERENCES

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UV-VIS SPECTROSCOPY 1.1 INTRODUCTION OF UV-VIS SPECTROSCOPY

For the UV-VIS Spectroscopy, it is concern about the interactions of radiation with matter which used to obtain all kind of information. According to the Prof Greenlief (2004), the absorption of light by solution is the oldest but it still one of the more useful instruments methods. Thus UV-VIS spectrometer is included as one of the oldest instrumental techniques of analysis and it also the basis of the ideal method that can analyzed the quantities and qualitative of the sample. In the UV-VIS, the absorption of radiation is dependent on the electronic structure of the absorbing species such as ion, atom and molecules. The matter can absorb the radiation by following transition and by this kind of transition it will raise the internal energy of the matter. The increase in energy equal to the energy absorb by the matter. Rotational : the molecule rotates about various axes, the energy of rotational being definite energy levels, so the molecule may absorb radiation and be raised to a higher rotational energy level. Vibrational : the atoms or group of atoms within the molecule is vibrate with each other and thus energy is definite absorb. So that, the amount of energy is raised to a higher vibrational energy level. Electron : both atoms and molecule can undergo this transition and the electrons is transfer from one orbital to another.

By the absorb radiation, it will transform into the UV spectrum. The spectrum is different with different conditions which are gas phase, liquid phase and aqueous solution as figure 4.

Figure 4

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THEORY AND PRINCIPLES According to Wikipedia, the molecules that containing -electron or non- bonding electron which is lone pair (n-electron), it can absorb the energy in the form of UV to excite the electrons to the higher anti-bonding molecular orbitals. It can be concluded that the more easily the electron to the excited level when the wavelength of the light it absorb is longer. The energy absorb by the molecules can change the between the ground state and excited state of electrons within the system. The electron can be classes into 3 types namely (single bond), (multiple bond) or non bonding (n-lone pair). Thus the electron when converted into energy of radiation it can excited from highest occupied molecular orbital (HOMO) to the lowest unoccupied molecular orbital (LUMO). Most of the UV-VIS Spectro occur at the -* or n- * which occur above 200nm wavelength while for n-* it occur at the short wavelength. The figure 5 below shows the different transition between electronic states.

Figure 5

For the single bond compound, it has not been widely used for this analytical purpose because of their difficulties absorption of radiation. Thus it can be analyzed for the compound with the double or triple bonds because they not held strongly as single bond. Then, these kinds of molecules are easily excited by the radiation. According to Skoog (2004),the unsaturated organic functional group which absorb the ultraviolet are known as the chromophores as figure 6.

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Spectra changes can be classed into : Bathochromic shift (red shift)- shift to longer wavelength Hypsochromatic shift (blue shift) shift to shorter wavelength Hyperchromatic effect - shift to greater absorbance Hypochromatic effect shift to shorter absorbance

INSTRUMENT OF UV VIS SPECTROSCOPY For this kind of instrument, they are made up of 5 components which are: a) a stable sources of radiant energy b) a wavelength selector c) one or more sample container d) a radiation detector e) a signal processing and readout unit. The layout of the components in the uv-visible spectroscopy:

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a) Radiation source

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According to Skoog (2004),the output power should be stable for reasonable periods of time. Thus, to get the good stability, the power supply must be enough to provide electrical power to the sources. In the uv-vis spectroscopy, they widely used continuum sources for example is deuterium and hydrogen lamps that can emit radiation in the range of 160-380 nm. For both of this lamps, the quartz windows had been used and also quartz cuvette because of the glass that absorb radiation of wavelength less than 350nm.

b) Wavelength selector (filter/monochromators) For the monochromator, there are following components ;

c) Sample container According to the Instrumentation of uv in pharmatutor, there are many kinds of sample cell that used in the uv which based on the: 1. The path length, shape, size 2. The transmission characteristics at the desired wavelength 3. The relative expense The cuvette that used for the sample should be transparent so that the light can passed through into it and the result will be recorded. For the uv, quartz and fused silica cuvette are used based on the wavelength which is range between 180-2500nm. The thickness generally is 1cm and rectangular in shape.

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d) Detector To detect the radiation, there are 3 types of photosensitive devices that are used which are 1. Photovoltaic or barrier layer cell 2. Phototubes or photoemissive tubes 3. Photomultiplier tubes

ULTRAVIOLET-VISIBLE PHOTOMETERS AND SPECTROPHOTOMETERS Spectrophotometers are the instrument that is used to measure the wavelength distribution of light. It also used to characterize the color of objects.

SINGLE-BEAM INSTRUMENT According to Brian M. Tissue,2000 , this spectrophotometers are often sufficient for making quantitative absorption measuerements in the UV-Vis spectral region. By measuring the absorbance at a single wavelength and applying the Beer-lambert Law the concentration of an analyte in solution can be determined. This spectrophotometers can utilize a fixed wavelength light source or a continuous source. The simplest instruments use a single-wavelength light source: Light-emitting diode (LED) A sample container Photodiode detector

Instruments that contain continuous source have a dispersing element and aperture or slit to select a single wavelength before the light passes through the sample cell.

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To obtain the transmittance reading: 1) Digital readout is zeroed with the sample compartment empty so that the occluder (vane that is automatically falls between the beam and the detector whenener the cylindrical cell is removed from its holder) blocks the beam and no radiation reaches the detector. Called as 0% calibration process or adjustment. 2) Insert cell containing the blank(often the solvent) into the cell holder and the pointer is brought to the 100% T mark by adjusting the position of the light control aperture and thus the amount of light reaching the detector. Called as 100% T calibration or adjustment. 3) Place the sample in the cell compartment and the percent transmittance or the absorbance is read directly from the LED readout.

Spectral range of spectronic 20 : 340-950 nm Spectral band-pass: 20 nm Wavelength accuracy : 2.5 nm Photometric accuracy : 2%T

DOUBLE BEAM INSTRUMENT The purpose of double beam instrument is to determine the amount of light of a specific wavelength absorbed by an analyte in a sample. There are 2 types of double-beam instruments:

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1) Dual beam in space

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The beam are separated by beam splitter(V-shape). One beam passes through the reference solution to a photodetector, and the second simultaneously passes through the sample to a second matched photodetector. The two outputs are amplified, and their ratio, is obtained electronically or computed and displayed on the output device 2) Dual beam in time The beam are separated by in time rotating sector mirror. It directs the entire beam through the reference cell and then through the sample cell. The pulses of radiation are then combined by another mirror, which is it transmits the reference beam and reflects the sample beam to the detector. The dual beam in space is an instrument which is has two matched detectors. While the dual beam in time is the instrument which is uses a beam copper to automatically monitor the sample and reference signals sequentially with short intervals between measurements. Double-beam in time is generally more preferred compare to the double-beam in space. This is due to the difficulty in matching two detectors. The disadvantages of double beam instruments: The light path must be optically equal and have similarly efficient optical components. If the mirror used, it need to be coated at the same time. The mirror is need to be replaced in pairs. Build up of dust or film over the time may affect the beams. The requirements may make dual beam instruments complicated, difficult to align, and expensive to service.

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2.0 FOURIER-TRANSFORM SPECTROSCOPY INTRODUCTION FTIR is Fourier Transform Infrared Spectrometer. It is the preferred method of infrared spectroscopy. An FTIR consist of a single beam optical assembly with, as an essential component, an interferometer and the sample. It have three active components, which are a moving mirror, a fixed mirror and a beam splitter (Chemical Analysis, 2007).

In infrared spectroscopy, infrared (IR) radiation will pass through a sample. Some of the radiation will be absorbed by the sample and some of it will be trasmitted. The resulting spectrum will show the molecular absorption and transmission, creating a molecular fiengerprint of the sample. It is called a fingerprint because there will never have two or more unique molecular structures that will produce the same infrared spectrum. That is the reason that makes the infrared spectroscopy very useful for some analysis (Thermo Nicolet Corporation, 2001). HISTORY The first generation infrared spectrometer was invented in late 1950s. It utilizes prism optical splitting system. The prism are made of NaCL, the requirement of the sample is very strict and the scan range is narrow. So, it is no longer being used. The second generation was introduced in 1960s. It utilizes gratings as the monochrometer. The performance are much better than the first generation infrared spectrometer but there are still several prominent weaknesses. The invention of the third generation infrared spectrometer, FTIR spectrometer, marked the abdication of monochrometer an dthe prosperity of interferometer (UC Davis Chemwiki, 2010). The Infrared Spectroscopy has been a very useful technique for materials analysis in the laboratory for over seventy years. During the past years, the technique of FTIR has

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become more important as an instrument to obtain a high quality infrared spectra in analytical research and process control. There are some reasons for this development, one of it is the capability of FTIR spectrometer itself. The FTIR can produce routine spectra in much shorter times with better quality than those from conventional grating system and the extension of the possibilities of performing measurements which would be impossible with grating system. The FTIR spectrometer also can produce rapid sample measurement, absolute wavelength accuracy, versatile spectra processing and total automation of compex analysis (Indian Institute of Technology Bombay) The original infrared instruments were of the dispersive type. These instruments will separate the individual frequencies of energy emitted from the infrared source. This was accomplished by the use of a prism or grating. An infrared prism works exactly the same as a visible prism which separates visible light into its frequencies. A grating is a more modern dispersive element which better in separates the frequencies of infrared energy. The detector will measure the amount of energy at each frequency which has passed through the sample. This results in a spectrum which is a graph of intensity vs frequency (Thermo Nicolet Corporation, 2001). Principles of Fourier Transfer Infrared Spectroscopy Figure 1 shows the basic components of an FTIR. The infrared source emits a broad band of different wavelength of infrared radiation. The IR (Infrared) source used in the Temet GASMET FTIR CR-series is a SiC (Silicon Carbide) ceramic at a temperature of 1550 K. The IR radiation goes through an interferometer that adjusts the infrared radiation. The interferometer performs an optical inverse Fourier transform on the entering IR radiation. The modulated IR beam passes through the gas sample where it is absorbed to various extents at different wavelengths by the various molecules present. Finally the intensity of the IR beam is detected by a detector, which is a liquid-nitrogen cooled MCT (MercuryCadmium-Telluride) detector in the case of the Temet GASMET FTIR CR-series. The detected signal is digitised and Fourier transformed by the computer to get the IR spectrum of the sample gas. (Basil Daham,UK)

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Figure 1: Basic components of FTIR (Basil Daham, UK)

A SIMPLE SPECTROMETER LAYOUT

Figure 2 : FTIR spectrometer (Thermo Nicolet Corporation, 2001) In addition,the unique part of an FTIR spectrometer is the interferometer. Figure 3 shows a Michelson type plane mirror interferometer. Infrared radiation from the source is collected and collimated (made parallel) before it strikes the beam splitter. The beam splitter ideally transmits one half of the radiation, and reflects the other half. Both transmitted and reflected beams strike mirrors, which reflect the two beams back to the beam splitter. Thus, one half of the infrared radiation that finally goes to the sample gas has first been reflected from the beam splitter to the moving mirror, and then back to the beam splitter. The other half of the infrared radiation going to the sample has first gone

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through the beam splitter and then reflected from the fixed mirror back to the beam splitter. When these two optical paths are reunited, interference occurs at the beam splitter because of the optical path difference caused by the scanning of the moving mirror. (Basil Daham, UK)

Figure 3: Michelson interferometer (Basil Daham, UK)

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3.0 REFERENCES:

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1. D.Skoog, D.M.West, F.J.Holler, Fundamental of Analytical Chemistry, Eight Editions, Instrument of Optical Chemistry, Canada, 2004. 2. http://www.pharmatutor.org/pharma-analysis/analytical-aspects-of-uv-visiblespectroscopy/instrumentation-of-absorption-spectroscopy.html 3. http://en.wikipedia.org/wiki/Ultraviolet%E2%80%93visible_spectroscopy 4. Dr Hinna Hamid, Ultraviolet and Visible Spectrometry (12.07.2007) 5. Prof Greenlief, Uv-Vis Absorption Spectroscopy, 2004 6. Daham, B. (n.d.). Principles of Fourier Transfer Infrared Spectroscopy. Retrieved march 17, 2013, from http://www.daham.org/basil/leedswww/ftir/ftir_principle.htm 7. Thermo Nicolet Corporation. (2001). Introduction to Fourier Transform Infrared Spectrometry, 6. 8. http://www.rose-hulman.edu/~brandt/Fluorescence/Absorbance_Spectroscopy.pdf (Mark Brandt,1999-2010) 9. http://www.shsu.edu/~chm_tgc/primers/spect.html (Dr. Thomas G. Chasten,1998,2007,2009) 10. http://www.hunterlab.com/appnotes/an11_97.pdf 11. http://www.files.chem.vt.edu/chem-ed/spec/uv-vis/singlebeam.html (Brian M. Tissue,2000) 12. (Fundamental of analytical chemistry;Brooks/Cole,Cengange Learning;Skoog,West,Holler,Crouch;2004) 13. Rouessac, Francis & Rouessac, A (2007). Chemical Analysis. French: John Wiley & Sons 14. (2001). Introducton to fourier Transform Infrare Spectrometry. Thermo Nicolet Corporation, . retrieved March, 16 2013 , from http://www.google.com.my/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0C EUQFjAA&url=http%3A%2F%2Fmmrc.caltech.edu%2FFTIR%2FNicolet%2FNicolet %2520Software%2FNicolet%25202%2F4700_6700_User.pdf&ei=zPtFUbSVGoTTrQ eIuIHICg&usg=AFQjCNH3XVONAAeR7T3mUV1Kk5ckaTQJNg 15. Fourier Transform Infrared Spectrometer. (n.d) retrieved March, 16 2013, from Indian Institute of Technology, Bombay Web Site: http://www.rsic.iitb.ac.in/Ftir.html 16. How an FTIR Spectrometer Operates. (2010). retrieved March, 16 2013, from Uc Davis ChemWiki Web Site: http://chemwiki.ucdavis.edu/Physical_Chemistry/Spectroscopy/Vibrational_Spectro scopy/Infrared_Spectroscopy/How_an_FTIR_Spectrometer_Operates?highlight=FT IR

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