Madhavi mutha


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Madhavi mutha

  1. 1. A Seminar On Significance Of Separation Techniques In Pharmaceutical Analysis Presented By, Guided By, Miss. Madhavi Mutha Dr. Rajesh J Oswal Prof.Sandip Kshirsagar Department of Pharmaceutical Chemistry6/13/2012 1
  2. 2. INTRODUCTION . Separations touch every branch of science and technology and have developed into a well established branch known as separation science. If you just look at the developments, say in biological sciences such as biochemistry, biotechnology, clinical pharmacology, therapeutics and toxicology, the progress has taken place prominently because of the advancements in the separation methods.6/13/2012 2
  3. 3. SEPARATION METHODS − A UNIFIED SCIENCE The separations play a key role in the various branches of science and technology but they themselves form a unified branch of science. In the simplest terms, separation is defined as an operation in which a mixture is divided into at least two components having different compositions. But this particular definition has a limitation as it does not cover chiral separations in which molecules of same composition and chemical structure are involved. Therefore, a broader definition of separation will be as:“Separation is a process by which a mixture is divided in at least two components with different compositions or two types of molecules with the same composition but different stereochemistry.”6/13/2012 3
  4. 4. Objectivesi) Analysis of different constituents of a mixture.ii) Procuring pure materials from complex mixtures. In analysis, (i), there may be two aspects:• Removal of interfering constituents before the actualquantitation of one or more known compounds.• Isolation of unknown constituents for subsequentcharacterization.Under “(ii)”, for obtaining the pure materials fromcomplex mixtures, the constituent with the desired purity.In some cases, in order to attain the required level ofpurity, the separation steps of the same process mayhave to be repeated.6/13/2012 4
  5. 5. SIGNIFICANCE OF SEPARATION TECHNIQUES Analytical chemistry:1.Classical methods:. Qualitative analysisA qualitative analysis determines the presence or absence of a particularcompound, but not the mass or concentration..Flame test.Inorganic qualitative analysis generally refers to a systematic scheme toconfirm the presence of certain, usually aqueous, ions or elements byperforming a series of reactions that eliminate ranges of possibilities and thenconfirms suspected ions with a confirming test.. 6/13/2012 5
  6. 6. Gravimetric analysis:Gravimetric analysis involves determining the amount of materialpresent by weighing the sample before and/or after sometransformation. A common example used in undergraduate educationis the determination of the amount of water in a hydrate by heating thesample to remove the water such that the difference in weight is dueto the loss of water..Volumetric analysis:Titration involves the addition of a reactant to a solution beinganalyzed until some equivalence point is reached. Often the amountof material in the solution being analyzed may be determined. 6/13/2012 6
  7. 7. 2.Instrumental methods:.Spectroscopy:Spectroscopy measures the interaction of the molecules withelectromagnetic radiation. Spectroscopy consists of manydifferent applications such as atomic absorption spectroscopy,atomic emission spectroscopy, ultraviolet-visible spectroscopy,x-ray fluorescence spectroscopy, infrared spectroscopy, Ramanspectroscopy, dual polarisation interferometry, nuclear magneticresonance spectroscopy, photoemission spectroscopy,Mössbauer spectroscopy and so on 6/13/2012 7
  8. 8. .Mass spectrometry :Mass spectrometry measures mass-to-charge ratio of moleculesusing electric and magnetic fields.Electrochemical analysis:Genomics - DNA sequencing and its related research. Geneticfingerprinting and DNA microarrayProteomics - the analysis of protein concentrations andmodifications, especially in response to various stressors,at variousdevelopmental stages, or in various parts of the body.Metabolomics - similar to proteomics, but dealing with metabolites.Transcriptomics - mRNA and its associated fieldLipidomics - lipids and its associated fieldPeptidomics - peptides and its associated field6/13/2012 8
  9. 9. .Thermal analysis:Calorimetry and thermogravimetric analysis measure theinteraction of a material and heat..Separation: Adsorption, adhesion of atoms, ions or molecules of gas, liquid,or dissolved solids to a surface..Centrifugation: Separating textile. Removing water from lettuce after washing it in a salad spinner. Separating particles from an air-flow using cyclonic separation. The clarification and stabilization of wine. 6/13/2012 9
  10. 10. Chromatography:Column chromatography :Column chromatography is a separation technique in which thestationary bed is within a tube. The particles of the solid stationaryphase or the support coated with a liquid stationary phase may fill thewhole inside volume of the tube (packed column) or be concentratedon or along the inside tube wall leaving an open, unrestricted path forthe mobile phase in the middle part of the tube (open tubular column).Differences in rates of movement through the medium are calculatedto different retention times of the sample.Paper chromatography:.separation and identification of mixtures that are or can becolored, especially pigments.Two-way paper chromatography, involves using two solvents androtating the paper 90° in between. This is useful for separatingcomplex mixtures of similar compounds, for example, amino acids.6/13/2012 10
  11. 11. .Thin layer chromatography: .analyzing ceramides and fatty acids. .detection of pesticides or insecticides in food and water. .analyzing the dye composition of fibers in forensics. .assaying the radiochemical purity of radiopharmaceuticals..Displacement chromatography:.purification of proteins from complex mixtures..Two-dimensional chromatography represents rigorous approach toevaluation of the proteome. 6/13/2012 11
  12. 12. .Gas chromatography: Gas chromatography (GC), is used in analysing compounds that can be vaporized without decomposition. It is well suited for use in the petrochemical, environmental monitoring and remediation, and industrial chemical fields. .Liquid chromatography: High-performance liquid chromatography: HPLC, is a chromatographic technique that can separate a mixture of compounds and is used in biochemistry and analytical chemistry to identify, quantify and purify the individual components of the mixture.6/13/2012 12
  13. 13. .Affinity chromatography:.Purify and concentrate a substance from a mixture into a bufferingsolution.Reduce the amount of a substance in a mixture.Discern what biological compounds bind to a particular substance.Purify and concentrate an enzyme solution..Supercritical fluid chromatography:(SFC) is a form of normal phase chromatography that is used forthe analysis and purification of low to moderate molecular weight,thermally labile molecules. It can also be used for the separation ofchiral compounds .Ion-exchange chromatography:It has significance in water purification, preconcentration of tracecomponents, ligand-exchange chromatography, ion-exchangechromatography of proteins, high-pH anion-exchangechromatography of carbohydrates and oligosaccharides, and others.6/13/2012 13
  14. 14. .Size-exclusion chromatography:The main application of gel-filtration chromatography is thefractionation of proteins and other water-soluble polymers, while gelpermeation chromatography is used to analyze the molecular weightdistribution of organic-soluble polymers.Hybrid techniquesGas chromatography–mass spectrometry:GC-MS include drug detection, fire investigation, environmental analysis,explosives investigation, and identification of unknown samples.GC-MS has been widely heralded as a "gold standard" for forensicsubstance identification because it is used to perform a specific testNuclear magnetic resonance spectroscopy:NMR spectroscopy is often the only way to obtain high resolutioninformation on partially or wholly unstructured proteins."Nucleic acid NMR" is the use of NMR spectroscopy to obtaininformation about the structure and dynamics of polynucleic acids,such as DNA or RNA.6/13/2012 14
  15. 15. Infrared spectroscopy: Used in the long-term unattended measurement of CO2concentrations in greenhouses and growth chambers by infrared gas analyzers. It also has significance in forensic analysis Infrared spectroscopy has also been successfully utilized in the field of semiconductor microelectronics.Liquid chromatography–mass spectrometry: LC-MS is very commonly used in pharmacokinetic studies of pharmaceuticals and is most frequently used technique in the field of bioanalysis and Metabolics. LC-MS is frequently used in drug development at many different stages including Peptide Mapping, Glycoprotein Mapping, Natural Products Dereplication, Bioaffinity Screening, In Vivo Drug Screening, Metabolic Stability Screening, etc.6/13/2012 15
  16. 16. CONCLUSION: : It deals with separation of mixtures to enhance purity of substance. Separation techniques are important because most substances are needed in the pure state.6/13/2012 16
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