FTIR ANALYSIS OF DICLOFENAC SODIUM

Project work submitted in partial fulfillment of the requirement for the award of the Degree of

MASTER OF SCIENCE In PHYSICS by N. SAMPATH KUMAR

REGISTER NO: 2270734

DEPARTMENT OF PHYSICS DWARAKA DOSS GOVERDHAN DOSS VAISHNAV COLLEGE CHENNAI-600 106 APRIL 2010

BONAFIDE CERTIFICATE

This is to certify that the report entitled FTIR ANALYSIS OF DICLOFENAC SODIUM being submitted to the Madras University, Chennai, by Mr.N.SAMPATH KUMAR, for the partial fulfillment for the award of degree of M.Sc., (Physics) is a bonafied record of work carried out by him, under my guidance and supervision.

Place: Date :

DECLARATION

I here by declare that this dissertation entitled FTIR ANALYSIS OF DICLOFENAC SODIUM is based on the original work done by me under the guidance and supervision of Mr.L.T.V.Vasanthagopalan, Lecturer, Department of Physics, D.G. Vaishnav College, Chennai-600 106 and has not formed the basis for the award of any degree, diploma, associateship or similar titles.

Place: Date:

N.SAMPATH KUMAR Reg. No: 2270734 D.G. Vaishnav College, Chennai-600 106.

ACKNOWLEDGEMENT
I gratefully acknowledge

Dr.S.Narasimhan,

Principal,

D.G.Vaishnav College, Chennai-600 106, for his kind encouragement throughout my course. I wish to thank Dr.D.Uthra, Head, Department of Physics, D.G.Vaishnav College, Chennai- 600 106, for her support throughout my course. My sincere thanks to Dr. B.Anita and Prof. B.Sylaja, Department of Physics, D.G.Vaishnav College, Chennai - 600 106. I wish to express my sincere thanks Department to my of guide Physics,

Mr.L.T.V.Vasanthagopalan,

Lecturer,

D.G.Vaishnav College, Chennai-600 106, who has suggested and guided me to select this project and shown keen interest during the course of the project. I cordially thank my friend encouragement to my project work. Mr.Shanmugapriyan, for his

Date : Place: (N.SAMPATH KUMAR)

CONTENTS
CHAPTER 1 INTRODUCTION 1

1.1 1.2 1.3 1.4 1.5 1.6 1.7

Introduction to Spectroscopy Fourier Transform Infrared spectroscopy (FTIR) Diclofenac Sodium Phototoxicity and Photosensitivity Ecological problem FTIR study Scope of study INSTRUMENTATION OF FTIR

1 2 4 9 13 14 15 16 16 17 22 28 29 29 31 38

CHAPTER 2 2.1 2.2 2.3 2.4 2.5 2.6

Infrared spectroscopy Theory of infrared absorption FTIR spectroscopy Description of FTIR spectrophotometer Applications of FTIR Advantages of FTIR

CHAPTER 3 RESULTS AND DISCUSSION 3.1 Conclusion

LIST OF TABLES 1 Infrared region and corresponding wave numbers 16

2 Vibrational frequencies for Normal and Sunlight condition 3 Vibrational frequencies for Normal and ice point condition LIST OF FIGURES 1 2 3 4 5 Chemical Structure of Diclofenac Sodium Optical geometries of two conformational isomers Photodegradation of Diclofenac sodium Bending and Stretching vibrations Optical diagram of FTIR spectrophotometer

34 37

4 6 12 19 24

CHAPTER 1
1.1 INTRODUCTION TO SPECTROSCOPY Spectroscopy is studying the properties of matter through its interaction with different frequency components of the electromagnetic spectrum. Although almost all parts of the electromagnetic spectrum are

used for studying matter in organic chemistry, we are mainly concerned with energy absorption from three or four regions: Ultraviolet and visible; infrared (IR); microwave; and radio frequency absorption. Infrared spectroscopy is the subset of spectroscopy that deals with the infrared region of the electromagnetic spectrum. IR spectroscopy is basically vibrational spectroscopy. It covers a range of techniques, the most common being a form of absorption spectroscopy. As with all spectroscopic techniques, it can be used to identify compounds and investigate sample composition. A common laboratory instrument that uses this technique is an infrared spectrophotometer. The infrared portion of the electromagnetic spectrum is usually divided into three regions; the near-, mid- and far- infrared, named for their relation to the visible spectrum. The far-infrared, approximately 400-10 cm -1 (1000-30 m), lying adjacent to the microwave region, has low energy and may be used for rotational spectroscopy. The mid-infrared, approximately 4000-400 cm-1 (30-2.5 m) may be used to study the fundamental vibrations and associated rotational-vibrational structure. The higher energy near-IR, approximately 14000-4000 cm-1 (2.5-0.8 m) can excite overtone or harmonic vibrations. The names and classifications of these subregions are merely conventions. They are neither strict divisions nor based on exact molecular or electromagnetic properties. The infrared spectrum of a sample is recorded by passing a beam of infrared light through the sample. Examination of the transmitted light reveals how much energy was absorbed at each wavelength. This can be

done with a monochromatic beam, which changes in wavelength over time, or by using a Fourier transform instrument to measure all wavelengths at once. From this, a transmittance or absorbance spectrum can be produced, showing at which IR wavelengths the sample absorbs. Analysis of these absorption characteristics reveals details about the molecular structure of the sample. When the frequency of the IR is the same as the vibrational frequency of a bond, absorption occurs. This technique works almost exclusively on samples with covalent bonds. Simple spectra are obtained from samples with few IR active bonds and high levels of purity. More complex molecular structures lead to more absorption bands and more complex spectra. The technique has been used for the characterization of very complex mixtures. 1.2 FOURIER TRANSFORM INFRARED SPECTROSCOPY (FTIR) FTIR is most useful for identifying chemicals that are either organic or inorganic. It can be utilized to quantitate some components of an unknown mixture. It can be applied to the analysis of solids, liquids, and gasses. The term Fourier Transform Infrared Spectroscopy (FTIR) refers to a fairly recent development in the manner in which the data is collected and converted from an interference pattern to a spectrum. Today's FTIR instruments are computerized which makes them faster and more sensitive than the older dispersive instruments. Fourier transform infrared (FTIR) spectroscopy is a measurement technique for collecting infrared spectra. Instead of recording the amount of

energy absorbed when the frequency of the infra-red light is varied (monochromator), the IR light is guided through an interferometer. After passing through the sample, the measured signal is the interferogram. Performing a Fourier transform on this signal data results in a spectrum identical to that from conventional (dispersive) infrared spectroscopy. 1.2.1 QUALITATIVE ANALYSIS FTIR can be used to identify chemicals from spills, paints, polymers, coatings, drugs, and contaminants. FTIR is perhaps the most powerful tool for identifying types of chemical bonds (functional groups). The wavelength of light absorbed is characteristic of the chemical bond as can be seen in this annotated spectrum. By interpreting the infrared absorption spectrum, the chemical bonds in a molecule can be determined. FTIR spectra of pure compounds are generally so unique that they are like a molecular "fingerprint". While organic compounds have very rich, detailed spectra, inorganic compounds are usually much simpler. For most common materials, the spectrum of an unknown can be identified by comparison to a library of known compounds. We have several infrared spectral libraries including on-line computer libraries. To identify less common materials, IR will need to be combined with nuclear magnetic resonance, mass spectrometry, emission spectroscopy, X-ray diffraction, and/or other techniques. FTIR spectrometers are cheaper than conventional spectrometers because building an interferometer is easier than the fabrication of a

monochromator. In addition, measurement of a single spectrum is faster for the FTIR technique because the information at all frequencies is collected simultaneously. This allows multiple samples to be collected and averaged together resulting in an improvement in sensitivity. 1.3 DICLOFENAC SODIUM

IUPAC Name: sodium (O-(2,6-dichloroanilino)phenyl)acetate. Molecular Formula: C14H10Cl2NNaO2. Molecular Weight: 318.1 g/mol.

Non-steroidal anti-inflammatory drugs (NSAIDs) comprise a large group of compounds that can be divided into two main sub-groups namely carboxylic acid and enolic acid sub-groups. Diclofenac sodium is a strong anti-inflammatory drug. Like other NSAIDs, diclofenac sodium has three basic properties-anti-inflammatory, antipyretic and analgesic. The antiinflammatory action is due to inhibition of prostaglandin (PG) synthesis, and preventing action of other mediators of inflammation locally. The agent enhances recovery by abolishing inflammation, diminishing pain and providing symptomatic relief and well being to the patient. Diclofenac sodium that consists of a phenylacetate group, a secondary amino group and a dichlorophenyl ring has limited water solubility

especially in gastric juice. A possibility to overcome this limitation is the complexation with -cyclodextrin. -cyclodextrin is a cyclic oligosaccharide consisting of seven glucopyranose units that can be represented as truncated cone structure with the wide and narrow rims occupied by the secondary and primary hydroxyl group, respectively. The central cavities of these molecules are hydrophobic and thus are able to encapsulate a wide variety of molecules. 1.3.1 PHARMACOLOGY The exact mechanism of action is not entirely known, but it is thought that the primary mechanism responsible for its anti-inflammatory, antipyretic, and analgesic action is inhibition of prostaglandin synthesis by inhibition of cyclooxygenase (COX) and it appears to inhibit DNA synthesis. Inhibition of COX also decreases prostaglandins in the epithelium of the stomach, making it more sensitive to corrosion by gastric acid. This is also the main side effect of diclofenac sodium. It has a low to moderate preference to block the COX-2-isoenzyme (approximately 10-fold) and is said to have therefore a somewhat lower incidence of gastrointestinal complaints than noted with aspirin.

Optimized geometries of two conformational isomers of Diclofenac sodium

There are two isomers of diclofenac sodium. Conformer-2 isomer of diclofenac sodium is more stable than the other isomer. The biological halflife of diclofenac sodium is about 1-2h, therefore it requires multiple dosing to maintain therapeutic drug blood level. 1.3.2 INDICATIONS Diclofenac sodium is used for musculoskeletal complaints, especially arthritis, rheumatoid arthritis, Polymyositis, Dermatomyositis, osteoarthritis,dental pain, spondylarthritis, ankylosing spondylitis, and pain management in cases of kidney stones and gallstones. An additional indication is the treatment of acute migraines. Diclofenac sodium is used commonly to treat mild to moderate post-operative or post-traumatic pain, particularly when inflammation is also present, and is effective against menstrual pain and endometriosis. Diclofenac sodium is often used to treat chronic pain associated with cancer, particularly if inflammation is also present.

1.3.3 SIDE EFFECTS

*

Cardiac Following the identification of increased risks of heart attacks with the

selective COX-2 inhibitor, it is concluded that diclofenac sodium does increase the risk of myocardial infarction.
*

Gastrointestinal Gastrointestinal complaints are most often noted. The development of

ulceration and/or bleeding requires immediate termination of treatment with diclofenac sodium. Most patients receive an ulcer-protective drug as prophylaxis during long-term treatment.
*

Hepatic Liver damage occurs infrequently, and is usually reversible. Hepatitis

may occur rarely without any warning symptoms and may be fatal. Patients with osteoarthritis more often develop symptomatic liver disease than patients with rheumatoid arthritis. Liver function should be monitored regularly during long-term treatment. If used for the short term treatment of pain or fever, diclofenac sodium has not been found to be more hepatotoxic than other NSAIDs.
*

Renal Studies showed that diclofenac sodium caused acute kidney failure in

sensitive persons or animal species, and potentially during long term use. In non-sensitive persons resistance to side effects decreases with age. However,

diclofenac sodium appears to have a different mechanism of renal toxicity. 1.3.4 FORMULATION Diclofenac sodium is available in stomach acid resistant formulations (25 and 50 mg), fast disintegrating oral formulations (25 and 50 mg), slowand controlled-release forms (75, 100 or 150 mg), suppositories (50 and 100 mg), and injectable forms (50 and 75 mg). However, the solubility of diclofenac sodium is very limited in aqueous solutions around physiological pH. Parenteral or injectable formulations of diclofenac sodium have therefore necessitated the use of solubilizing additives such as propylene glycol. 1.3.5 INTERACTIONS WITH DIETARY SUPPLEMENTS
*

Calcium Diclofenac sodium decreases the amount of calcium lost in the urine,

which may help prevent bone loss in postmenopausal women.

*

L-tryptophan Diclofenac sodium causes complex changes to L-tryptophan levels in

the blood, but the clinical implications of this are unknown. More research is needed to determine whether supplementation with L-tryptophan is a good idea for people taking diclofenac.
*

Lithium Lithium is a mineral that may be present in some supplements and is

also used in large amounts to treat mood disorders such as manic-depression. Diclofenac sodium may inhibit the excretion of lithium from the body, resulting in higher blood levels of the mineral. Since minor changes in lithium blood levels can produce unwanted side effects, diclofenac sodium should be used with caution in people taking lithium supplements.

1.4 PHOTOTOXICITY AND PHOTOSENSITIVITY Phototoxicity and photoallergy are the two major types of photosensitisation. Phototoxicity occurs more frequently than photoallergy and does not involve an immunological mechanism. Phototoxic reactions, which resemble severe sunburn and may even blister, are dose dependent for both drugs and sunlight. These reactions occur immediately after, or within a few hours of taking the drug and simultaneous exposure to radiation of the appropriate wavelength and will subside if the drug is withdrawn and/or if possible the patient avoids excessive exposure to sunlight. Phototoxicity may cause damage to cells by modification of certain targets such as DNA, lipids and/or amino acids and proteins. Drugs may be exposed to a variety of light conditions, including direct sunlight, filtered (window) sunlight and various artificial light sources during manufacture, storage, distribution and administration to the patient. Although the ultraviolet (UV) component of sunlight is considered the most problematic, exposure to fluorescent lighting for prolonged periods of time needs to be taken into consideration in terms of the spectral distribution of these light sources, which extends from 300nm to 3000nm. UV-C (200-

280nm), UV-B (280-320nm) and UV-A (320-400nm) comprise the three regions of the UV-spectrum. Although eliminated from the earths surface by absorption by molecular oxygen and ozone, UV-C, which may cause adverse effects, is present in artificial light sources such as germicidal lamps and welding arcs. Although the UV-A reaches the earths surface to a greater extent than that of UV-B, it is the UV-B which causes sunburn and skin cancer. While sunlight and fluorescent light sources have a high output in the visible region of the spectrum (400-800 nm), the significant output in the UV-region must be noted for the fluorescent light sources. Most drugs absorb UV-C, while UV-B is responsible for the photoreactivity of drugs in the presence of sunlight and UV-A in addition to being absorbed by DNA, may cause photosensitisation reactions. In order to cause a reaction, the UV radiation must penetrate, whether it is the drug in the formulation or in the patient. The former is dependant on the transparency of the packaging. In the patient, although UV radiation shorter than 320nm penetrates the stratum corneum of the Caucasian skin, it must reach the absorbing drug molecule in the peripheral blood capillaries in order to illicit a photochemical response. Studies on the photodegradation of drugs are relevant to the drug development process, because the photolysis products may have biological effects different from those of the parent compounds. This may explain, at least partially, the phototoxicity mechanism. Reports on cutaneous photosensitivity disorders provoked by new pharmaceuticals appear with increasing frequency.

A mechanistic study of the photodegradation is found in the recent literature. The major photoproducts of diclofenac sodium are carbazole derivatives (compounds 8Clcb and cb). Assays in vitro performed with Diclofenac sodium and the photoproducts show positive photoxicity only for 8Clcb. Likewise, the analysis of the photophysical data are interpreted considering the participation of the triplet state of 8Clcb. Thermodynamic considerations as well as an observed quenching of triplet state as the concentration of 8Clcb is increased lead to formulate the formation of an excimer from which a radical ion could be formed. Dehalogenation is followed to yield an aryl radical and chloride anion. In other words, irradiation with UVA light of the anti-inflammatory drug diclofenac sodium in aqueous buffer or methanol solution leads to sequential loss of both chlorine substituents and ring closure to carbazole-1acetic acid as the major product. Minor products result from substitution by the solvent. Diclofenac sodium, after photodehalogenation (removal of both chlorine atoms) takes place, the carboxyl group remains intact which allows the photoproduct to be readily excreted. Drug concentration needs to be taken into consideration in all photosensitivity reactions and this involves

the dosage of the drug concerned the higher the dose the higher the circulation concentration of the drug and the greater the potential for a photosensitivity reaction to occur. This needs to be taken into consideration when comparing the ability of diclofenac sodium to cause photosensitivity effects in patients.

When the drug (Aryl-Cl) is photolyzed in aqueous or alcoholic (ROH) solution, HCl is liberated and a mixture of reduction (Aryl-H) and substitution (Aryl-OR) products is obtained. This is exemplified by the photodegradation of diclofenac sodium as shown in figure. The photodechlorination occurs for these compounds more strongly in deoxygenated solution. Many drugs causing adverse photosensitivity effects contain chlorine atoms as substituents, with the photolabile nature of chlorine being shown in chloroaromatic compounds. Diclofenac sodium has been reported to cause phototoxicity effects in patients due to a free radical photodechlorination

process. The photodegradation of diclofenac sodium in aqueous solution involved the loss of HCl to form the chlorocarbazole with subsequent loss of the second chlorine on further irradiation.

1.5 ECOLOGICAL PROBLEM

The presence of diclofenac sodium has been reported in natural waters and in wastewater treatment plant effluents as a consequence of its incomplete elimination with conventional wastewater treatments. . Diclofenac sodium enters the environment either as the parent compound or as active/inactive metabolites. Due to their pharmacological activity, their determination and understanding their behavior in the environment are important. Direct photolysis has been described as the main transformation pathway for its elimination under environmental conditions and some photoproducts have been identified as responsible for its phototoxicity.

1.6 FTIR STUDY FTIR spectra of diclofenac sodium, at three different storage conditions were obtained using a ABB BOMEM MB series 104 model spectrophotometer. 1.7 SCOPE OF STUDY COMPARATIVE STUDY OF DICLOFENAC SODIUM SALT AT DIFFERENT STORAGE CONDITIONS USING FTIR TECHNIQUE FTIR technique has been employed to determine whether there is any change in the potency of a drug if it is stored in different storage conditions. Three samples of diclofenac sodium salt (pure form) were taken. The first sample was stored in the normal condition. The second sample was freezed at -18C for 24 hours. The third sample was exposed to sunlight for 50 hours. FTIR spectra for all the samples were taken. FTIR spectra of diclofenac sodium at ice point condition and sunlight exposed condition were compared with the spectrum of the same drug at normal condition. The study of photolysis of diclofenac sodium in solid-state is of particular interest. Since diclofenac sodium as any other NSAID is known for phototoxicity, it will be useful if we study its photoproducts after exposure to sunlight. In present investigation, FTIR spectrum of diclofenac sodium at ice point is compared with the spectrum of the same drug at standard condition. On comparing, we concluded that freezing has no direct effect on the

efficacy of the given drug in solid state. Again the FTIR spectrum of diclofenac sodium at sunlight exposed condition is compared with the spectrum of the same drug at standard condition. On comparing, we can observe some changes in the spectrum of diclofenac sodium at sunlight condition. Some peaks were missing and new stretching frequency (broad band) was located in the range 2800-3600 cm -1, may be due to the overlapping CH and NH bands. It is inferred that different storage conditions would degrade the chemical potency of the drug.

CHAPTER 2

2.1 INFRA RED SPECTROSCOPY Infrared radiation cannot be seen by the human eye. This is because it lies outside the visible spectrum it has a lower energy. Although it is invisible, infrared radiation makes a highly visible contribution to physics and analytical chemistry. Infrared spectroscopy is one of the most important analytical techniques available to chemists and it is one of the most familiar. Infrared spectroscopy is a method of identifying substances by the infrared wavelengths they can absorb. Infrared light is on the spectrum just after visible light: The term infra red covers the range of the electromagnetic spectrum between 0.78 and 1000m. In the context of infrared spectroscopy, wavelength is measured in wave numbers, which have the units cm-1. Wave number = 1 / wavelength in centimeters. It is useful to divide the infra red region into three sections; near, mid and far infra red; The most useful I.R. region lies between 4000-670 cm-1.
TABLE 1: INFRARED REGION AND CORRESPONDING WAVE NUMBERS

Regio n Near Middle Far

Wave length range (m) 0.78 2.5 2.5 50 50 1000

Wave number range (cm-1) 12800- 4000 4000- 200 200 - 10

2.2 THEORY OF INFRA RED ABSORPTION

IR radiation does not have enough energy to induce electronic transitions as seen with UV. Absorption of IR is restricted to compounds with small energy differences in the possible vibrational and rotational states. Only those vibrations that result in the change of molecular dipole (separation of positive and negative charges) and having absorption frequencies in the infrared region of the spectrum absorb infrared radiation. Simple gas molecules such as H2, CI2 and O2 do not have dipoles and therefore do not give infrared spectra. However, molecules such as SO 2 and CO2 do have dipoles, hence they have infrared spectra. Atoms in molecules are not static, but rather they vibrate about their equilibrium positions. The frequency of these vibrations depends on the mass of the atom and the length and strength of the bonds. Molecular vibrations are stimulated by bonds absorbing radiation of the same frequency as their natural vibrational frequency (usually in the infrared region).
INTERACTION BETWEEN MOLECULAR DYNAMICS AND LIGHT

For a molecule to absorb IR, the vibrations or rotations within a

molecule must cause a net change in the dipole moment of the molecule. The alternating electrical field of the radiation interacts with fluctuations in the dipole moment of the molecule. If the frequency of the radiation matches the vibrational frequency of the molecule then radiation will be absorbed, causing a change in the amplitude of molecular vibration. Molecular rotations Rotational transitions are of little use to the spectroscopist. Rotational levels are quantized, and absorption of IR by gases yields line spectra. However, in liquids or solids, these lines broaden into a continuum due to molecular collisions and other interactions. Molecular vibrations The positions of atoms in a molecules are not fixed; they are subject to a number of different vibrations. Vibrations fall into the two main categories of stretching and bending.

Stretching : Change in inter atomic distance along bonds axis Bending : Change in angle between two bonds. There are four types of bend:
1 1 1 1

Rocking Scissoring Wagging Twisting

Vibrational Coupling In addition to the vibrations mentioned above, interaction between vibrations can occur (coupling) if the vibrating bonds are joined to a single, central atom. Vibrational coupling is influenced by a number of factors:
*

Strong coupling of stretching vibrations occurs when there is a

common atom between the two vibrating bonds

*

Coupling of bending vibrations occurs when there is a common bond between vibrating groups Coupling between a stretching vibration and a bending vibration occurs if the stretching bond is one side of an angle varied by bending vibration

Coupling is greatest when the coupled groups have approximately equal energies No coupling is seen between groups separated by two or more bonds Infrared spectroscopy has advanced rapidly as an analytical technique

and it can now be used to look at problems that were once thought to be too difficult. Protein structures can now be investigated as can microbiological systems- e.g. the identification of viruses and fungi. Infrared spectroscopy can also be used to study the particles that block air filters, how fats crystallize in margarines and low fat spreads, and it can also be used for categorizing or identifying fibers and paint chips in forensic analysis, and for the screening of illicit substances. The complexity of infrared spectra in the 1450 to 600 cm -1 region makes it difficult to assign all the absorption bands, and because of the unique patterns found there, it is often called the fingerprint region. Absorption bands in the 4000 to 1450 cm-1 region are usually due to stretching vibrations of diatomic units and this is sometimes called the group frequency region. The infrared spectrum can be split conveniently into four regions:

4000 2500 cm-1:

The absorption of single bonds to hydrogen - e.g. C-H, O-H and N-H: The high frequency of these stretches is due to low mass of the hydrogen atom. Typically C-H stretches are found here.
*

2500 2000 cm-1 :

The absorption of triple bonds e.g. CC and CN; Relatively high requencies are required to provide the high energies necessary to make the strong triple bonds vibrate e.g. CH3CH2CN.
*

2000 1500 cm-1 :

The absorption of double bonds e.g. C=C, C=O; This is an important region because it is where the C=O and C=C absorptions occur. These are important for identifying compounds containing these functional groups.
*

1500 - 400 cm-1 :

Absorption owing to other bond deformations e.g. rotating, scissoring and some bending. This region is known as the fingerprint region because this part of the spectrum is unique to each compound. This region is rarely used for identifying functional groups. Hydrogen bonding The absorption by O-H groups in alcohols and carboxylic acids normally gives broad bands because the vibrational mode is complicated by hydrogen bonding. The broadness depends on the degree of hydrogen bonding. The frequency of an alcoholic O-H stretch depends on whether the alcohol is primary, secondary or tertiary.

2.3 FTIR SPECTROSCOPY In Fourier transform infrared spectroscopy (FTIR) only one beam is used and all the required frequencies pass through the instrument at once. It is called FTIR spectroscopy because a mathematical treatment Fourier transformation is used to interpret the data and produce a spectrum. FTIR is a fast, sensitive technique and is now used in most modern chemical laboratories. Fourier Transform Infra-red (FTIR) Spectroscopy refers to an experimental technique for collecting Infra-red spectra. Instead of recording the amount of energy absorbed as the frequency of the infra-red light is varied, the sample is exposed to single pulse of infra-red light and the resulting signal observed. By performing a mathematical Fourier Transform on this response, a spectrum identical to that from conventional Infra-red spectroscopy can be derived. It can be applied to a variety of types of spectroscopy including optical spectroscopy, infrared spectroscopy (IR), nuclear magnetic resonance, and electron spin resonance spectroscopy. There are several methods of measuring the temporal coherence of the light, including the pulsed Fourier transform spectrograph and the continuous wave Michelson or Fourier transform spectrometer.

2.3.1 PRINCIPLE OF FTIR FTIR relies on the fact that the most molecules absorb light in the infra-red region of the electromagnetic spectrum, this absorption corresponds specifically to the bonds present in the molecule. The sample is

irradiated by a broad spectrum of Infra-red light and the level of absorbance at a particular frequency is plotted after Fourier transforming the data. The resulting spectrum is characteristic of the organic molecules present in the sample. In addition to standard transmission FTIR we also have the following sample handling capabilities:
*

Horizontal ATR attenuated total reflectance. Allows measurement of aqueous solutions, elastic and viscous samples which are difficult to grind

* *

Specular reflectance- allows measurements of thin films on metals KBr Disc allow suspension of powders in IR transparent KBr so they may be analysed Gas cells - This may be used for FTIR analysis of a gaseous sample (e.g. headspace analysis or environmental monitoring) Solution cells for measuring liquid sample in transmission mode

2.3.2 INSTRUMENTATION The Essential components of infrared spectrometer are

a) Infrared radiation source a) Monochromator a) Sample cells a) Detectors

a). Infrared radiation source The most commonly used light sources are nearest globar consist of a spindle of rare earth oxide (cesium, zirconium) of about 2.5cm long, 0.25cm diameter with a black body peak around 7100cm-1 at 1600 1700C. The globar is a low resistance silicon carbide with black body peak between 5500 500cm-1 at 1200C . It is more useful in longer wavelength. Ceramic globar consist of a mixture of zirconium oxide, yttrium oxide, ertium oxide heated at 1500C by electrical means. b). Monochromators Optical prism or gratings are used to obtain approximately

monochromatic component wavelength. Grating gives better resolving power. The resolution also depends on slit width and quality of mirrors. The rock salt prism is used as monochromator in the region 400 650cm-1. For higher resolution in a selected region of wavelength Lithium fluoride or Calcium fluoride are used. c). Sample cell The sample used may be gases, liquids, solids. The material containing the sample must be transparent to infrared radiation, for the sampling of solids any one of the method is employed. The solids are dissolved in solvents because solids result in scattering of infrared radiation too much. The absorption due to solvent has to be compensated by keeping the source in a cell of same thickness as that containing the sample in the reference beam. Solid films For solids of amorphous in nature, this technique is used. The sample is deposited on the surface of a potassium bromide or sodium chloride cell by evaporation of a solution of the solid. This technique used for rapid qualitative analysis only. Mull technique A Mull of the substance is prepared by grinding it into a fine powder and mixed with mulling agent such as paraffin oil to make a thick paste. This mull is spread between the two infrared transmitting windows and placed in the path of the infrared and the spectrum is run. For accurate measurements

the size of the sample particle be less than that of wavelength of radiation used. Pellet method In the technique a small amount of finely ground sample is intimately mixed with about 100 times its weight of potassium bromide, then it is passed under high pressure to form a small pellet of about 1-2nm thickness and 1cm in diameter. Sampling of liquids The sample in liquid at room temperature are taken into a rectangular cell made of sodium chloride or potassium bromide and thus infrared spectra are obtained directly. Sampling of gases The gas sample is put in a special cell of about 10cm long and is kept across the path of the infrared beam. The end walls of the cells are generally of sodium chloride. d). Detectors Infrared detectors are classified into two types. They are Thermal detectors and Photo detectors. Thermal detectors These detectors depend on the integrated energy of a large number of

incident photons to produce through their heating effects. Thermocouple is one of the most commonly used of thermal infrared detectors. Photo detectors Semiconductors devices in which an electron can absorb the energy of a single quantum of infrared radiation to be promoted from the valence band to the conduction band by which they can contribute to electrical conductivity. Photo detectors are not used in the laboratory spectrophotometer. Recorder The amplified signal is used to move an attenuator, which cuts down the radiation coming out of the reference beam until energy balance is restored. This is achieved by a motor which drives the comb into the reference beam. The recorder pen is also coupled to this motor so that the comb movement is followed exactly by the pen. Infrared spectrophotometer is of two types. They are a single beam spectrophotometer and a double beam spectrophotometer. The single beam instrument measures directly the amount of energy transmitted by the sample. It is the most accurate transmittance measurements. The double beam spectrophotometer is employed for radio recording of survey spectra covering a wide range double beam instrument is used in qualitative analysis. Most of the instrument is based on the double beam principle. High speed instrument are required for kinetic studies and for line identification of constituents.

2.4 DESCRIPTION OF FTIR SPECTROPHOTOMETER A Fourier transform spectrometer (abbreviated FTS) is a Michelson interferometer with a movable mirror. By scanning the movable mirror over some distance, an interference pattern is produced that encodes the spectrum of the source. A Fourier transform spectrometer consists of two mirrors located at a right angle to each other and oriented perpendicularly, with a beam splitter placed at the vertex of the right angle and oriented at a 45 angle relative to the two mirrors. Radiation incident on the beam splitter from one of the two ports is then divided into two parts, each of which propagates down one of the two arms and is reflected off one of the mirrors. The two beams are then recombined and transmitted out the other port. When the position of the mirror is continuously varied along the axis of the corresponding arm, an interference pattern is swept out as the two phaseshifted beams interfere with each other. Operation of infrared spectrometers Conventional infrared Spectrometers are double beam instruments. This means that there two radiation beams, one passing through the sample and the other passing through a reference which can be air. Radiation across the whole frequency range is passed through the sample. A monochromator is used to select radiation of only one frequency at a time to pass through the sample and the reference. As a particular frequency is absorbed by the sample less radiation is transmitted. A detector compares the energy that is transmitted by the sample with the radiation that is passing through the reference. From the difference the spectrum is then plotted.

2.5 TYPICAL APPLICATIONS OF FTIR

* * * *

Contamination detection Raw material testing Measurements of layers by specular and diffuse reflection Atmospheric Sampling using a portable unit Fourier transforms spectrometers. Aside from smelling tea, infra-red spectroscopy is widely used in both research and industry as a simple and reliable technique for measurement, quality control, and dynamic measurement. The instruments are now small, and can be transported, even for use in field trails.

2.6 ADVANTAGES OF FTIR The FTIR technique have many advantages over the plain infrared spectrometry in terms of scanning, signal to noise ratio, sensitivity, resolution and data processing.
(i)

The repaid scanning property has a great impact in the field of gas chromatography. Simultaneous data collection helps investigate spectrum of transient species such as unstable molecules or intermediate in a chemical regions.

(i)

(i)

Due to an increase of signal to noise ratio, the resolution is far superior to the ordinary spectrum. The sensitivity helps to investigate species on metal oxides, pharmaceuticals, proteins, etc.,

(i)

It make use all frequencies from the source simultaneously rather

than sequentially as in a scanning instrument.

(i)

The sensitivity of the Fourier transform method is greater than that of the dispersive technique because more radiation enters the slit less system. This is called Jacquinot advantage. FTIR spectrometers are cheaper than conventional spectrometers

because it is not necessary to vary the frequency of the infra-red light in a carefully controlled manner. In addition sampling time is smaller for the FTIR technique. This allows multiple samples to be collected and average together resulting in an improvement in sensitivity.

CHAPTER 3

RESULTS AND DISCUSSION

COMPARATIVE STUDY OF DICLOFENAC SODIUM AT THREE DIFFERENT CONDITIONS

a) Normal condition and Sunlight exposed condition White amorphous powder of Diclofenac sodium turned into yellow when it was exposed to sunlight for 50 hours. Corresponding FTIR spectrum was compared with the spectrum of diclofenac sodium at

normal condition. We can observe shifts in peaks of the sample at sunlight condition as compared with the spectrum at normal condition.

FTIR SPECTRUM OF DICLOFENAC AT NORMAL CONDITION

FTIR SPECTRA OF DICLOFENAC SODIUM EXPOSED TO SUNLIGHT

FINGERPRINT REGION OF DICLOFENAC SODIUM AT SUNLIGHT EXPOSED CONDITION

GROUP FREQUENCY REGION OF DICLOFENAC SODIUM AT SUNLIGHT EXPOSED CONDITION

Table 2: Vibrational frequencies for normal and sunlight exposed conditions

Assignment

Normal condition observed frequency (cm-1)

sunlight condition observed frequency (cm-1) 3367 3080 2971 1577 1284 and 1306 749 1665 to 2000 1453 1605 and 1471

aromatic secondary N-H

3387 3036 and 3079 2971 1575 1282 and 1305 747 1665 to 2000 1454 1604 and 1469

(m) aromatic C-H

aliphatic

C-H O=C-O- (vs)

asymmetric

C-N (s)
aromatic

C-Cl (s)
+

Overtones (weak bands)

metallic salt Na aromatic

C-C

The FTIR spectrum of the diclofenac sodium exhibited distinctive peak at 3387 cm-1 at normal condition and at 3367 cm-1 when exposed to sunlight due to N-H stretching of the secondary amine. And peaks at 3036 and 3080 cm-1 due to stretching of the aromatic C-H at normal condition and sunlight exposed condition respectively, and at 2971.41 cm-1 because of C-H aliphatic stretching at normal condition and sharp but weak peaks are visible correspondingly at sunlight exposed condition. The FTIR spectra of diclofenac sodium displayed very strong peak at 1575 cm-1 and 1577 cm-1 due to carboxyl group at normal condition and sunlight exposed condition respectively. And a strong peak at 747 cm -1 due to stretching of C-Cl bond. Sharp peaks at 1282 cm-1 and 1304 cm-1 due to C-N stretching of aromatic secondary amine at normal condition, whereas the corresponding peaks due to sunlight exposed condition appeared at 1284 cm -1 and 1306 cm-1 . Very broad band is observed in the range 2800 cm -1 to 3600 cm-1 may be due to overlapping of C-H and N-H bands. The absence of the carbonyl stretching band in the 18001600 cm1 spectral range confirms the presence of the carboxylate group in diclofenac sodium in solid state. b) Normal and ice point condition Diclofenac sodium salt was freezed at -18C for 24 hours.

Corresponding FTIR spectra was compared with the spectrum of diclofenac sodium which was maintained at normal storage condition. On comparing both the FTIR spectra no apparent changes were observed.

FTIR SPECTRUM OF DICLOFENAC SODIUM AT ICE POINT CONDITION

FINGERPRINT REGION OF DICLOFENAC SODIUM AT ICE POINT CONDITION

Table 3: Vibrational frequencies for normal and ice point conditions

Assignment

Normal condition observed frequency (cm-1)

Ice point condition observed frequency (cm-1) 3386 3035 and 3079 2971 1574 1282 and 1305 747 1665 to 2000 1454 1604 and 1468

aromatic secondary N-H

3387 3036 and 3079 2971 1575 1282 and 1305 747 1665 to 2000 1454 1604 and 1469

(m) aromatic C-H

aliphatic

C-H O=C-O- (vs)

asymmetric

CN (s)
aromatic

C-Cl (s)
+

Overtones (weak bands)

metallic salt Na aromatic

C-C

The FTIR spectra of the diclofenac sodium exhibited distinctive peak at 3387 cm-1 and 3386 cm-1 at normal and ice point conditions respectively, due to N-H stretching of the secondary amine. And peak at 3036 cm -1 and 3035 cm-1 due to stretching of the aromatic C-H at normal and ice point conditions respectively, and at 2971.41 cm-1 due to C-H aliphatic stretching. The FTIR spectra of diclofenac sodium displayed very strong peak at 1575 cm-1 and 1574 cm-1 due to carboxyl group at normal condition and ice point condition respectively. And a strong peak at 747 cm -1 due to stretching of CCl bond. Sharp peaks at 1282 cm-1 and 1305 cm-1 due to C-N stretching of aromatic secondary amine observed in both the conditions. The absence of the carbonyl stretching band in the 18001600 cm1 spectral range confirms the presence of the carboxylate group in diclofenac sodium in solid state.

CONCLUSION FTIR spectra of diclofenac sodium at different storage conditions were verified and it is concluded that freezing does not have any influence on the drug chemically. And this drug in the solid-state has undergone a colour change (reactions took place only in a thin surface-layer) on exposure to sunlight. This change in colour might not relate to chemical degradation of the drug; however the ability to effect changes in the physical stability such as tablet hardness, friability, disintegration and dissolution needs to be considered due to the potential changes in bioavailability and thus therapeutic efficacy. It is inferred that phototoxicity behaviour of diclofenac sodium cannot be verified by FTIR technique alone.

REFERENCES:

1. John VA. The pharmacokinetics and metabolism of diclofenac sodium (voltarol TM ) in animals and man.Rheumatol Rehabil Suppl, 2,1979, 22-27. 2. Gunasekaran S & Anita B, Indian J Pure & Appl Phys, 48(2010) 183-191 3. Gunasekaran S & Uthra D, Indian J Pure & Appl Phys, 46(Feb 2008) 100-105 4. Maitani Y, Kugo M, Nagai T. Permeation of diclofenac salt through silicone membrane: a mechanistic study of percutaneous absorption of ionizable drug. Chem Pharm Bull 42,1994, 1297-1301. 5. Fang, JY, Sung KC, Lin HH, Fang CL. Transdermal iontophoretic delivery of diclofenac sodium from various polymer formulations: in vitro and in vivo studies. Int. J. Pharm, 178, 1999,83-92 6. Kubitzek,F., Ziegler,G., Gold,M.S., Liu,J.H., and Ionescu,E. Low-dose diclofenac potassium in the treatment of episodic tension, European J.Pain 7(2), (2003) 155. 7. Jobin,F. and Gagnon,F.T. Inhibition of human platelet aggregation by a dibenzenazepine compound and by N-(2,6-dichlorophenyl)-o-aminophenylacetic acid, Canadian J.Phys.Pharm. 49, (1971) 479. 8. Sathyanarayana, D.N., Vibrational Spectroscopy; theory and applications New Age Int. (P) Ltd., (2004). 9. en.wikipedia.org/wiki/FTIR 10. www.freepatentsonline.com/5679660.html