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Group Members: Rubini Devi a/p Selvarajoo SB09005 Kalai selvi a/p Mohanraj SB09031

Group Members: Rubini Devi a/p Selvarajoo SB09005 Kalai selvi a/p Mohanraj SB09031 G.Sumithalaksmy a/p Gunasegara n SB09063 Calvin Lee Weng Leong SB09017.

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Group Members: Rubini Devi a/p Selvarajoo SB09005 Kalai selvi a/p Mohanraj SB09031

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  1. Group Members: • Rubini Devi a/p Selvarajoo SB09005 • Kalaiselvi a/p Mohanraj SB09031 • G.Sumithalaksmy • a/p Gunasegaran SB09063 • Calvin Lee Weng Leong SB09017

  2. Differentiate the extraction method used to extract metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.

  3. INTRODUCTION • Key problem of downstream processing methods is the high cost. • requires purification techniques that are delicate enough to preserve the biological activity. • In conventional methods, scale-up problems are enormous, making them uneconomical or very expensive.

  4. CONT…. • There is a need for efficient and economical approaches to the bioseparation problems. • Extraction using aqueous two-phase is one such possibility.

  5. Principle of Extraction Two kinds of polymer or a kind of polymer and a kind of salt dissolved in the same solvent, because of the Non-Intersoluble between polymers or polymer and salt, when the concentration of polymer or inorganic salt reached a certain value, will divided into two phase which are Non-Intersoluble. • Because all the solvent is water, so we call it aqueous two-phase, water hold a a large proportion in the two phase (85％一95％)

  6. Principle of Extraction Aqueous Two Phase Extraction is a method of extraction that utilize the materials which has got a different partition coefficient in two insoluble, aqueous two-phase. The system that can form aqueous two-phase are: Ion-type polymer - non-ion-type polymer(intermolecular repulsion) PEG－DEXTRAN polymer-low molecular weight compound(salting-out effect) PEG－Ammonium Sulfate

  7. Nucleic acid extraction methods: • Aqueous two-phase extraction • Phenol-choloroform extraction • Minicolumn purification • RNA extraction • Protein extraction methods • Aqueous two-phase extraction • Ion exchange chromatography • Size-exclusion chromatography • Affinity chromatography

  8. The Advantages of Aqueous Two-phase Extraction: • They provide mild conditions that do not harm or denature unstable/labile proteins and nucleic acids. • The polymer layer stabilizes the extracted protein and nucleic acid molecules, favouring a higher concentration of the desired protein in one of the layers, resulting in an effective extraction • The interfacial tension of two phase is low, magnitude order is 10-4N／cm, two phase are easy to disperse. • The proporation of two phase are changed by operating condition. • The density difference of the two phase is little, about 10 g/L. So it is not easy to separate the two phase, at present more research in this area. • Easy to continuous operation, big treatment capacity, was fitted for industry.

  9. Influencing Factors of Target Compound’s Partition Coefficient in Aqueous Two - phase System • The concentration of polymer- interfacial tension －the effect of interfacial tension to mass transfer is much greater than those other physical properties, for instance: density, viscosity, diffusion coefficient. • The relative molecular weight of polymer • General speaking, high molecular weight compounds(proteins) are easy to concentrated in low molecular weight phase. • electrochemical distribution • In aqueous two-phase extraction process, ionic strength shows little effect in partition coefficient of proteins. • hydrophobic reactions • biological affinity distribution • temperature and other factors

  10. Equipments Used • Mixer settler unit • Centrifuge tube • Centrifuge machine • Vortex

  11. PRINCIPLE OF CONVENTIONAL EXTRACTION SYSTEM • Low and intermediate molecular weightcompounds such as antibiotics, alkaloids, steroids and small peptides are generally extracted using aqueous/non-aqueous solvent system or also called conventional extraction system.

  12. Aqueous solvent system • Water is an excellent solvent but it has some limitations on its selectivity (it also extracts compounds of no interest along with the desired solute) • It normally requires further purifying processing such as ultrafiltration, drying and so on to obtain extracts of good quality.

  13. Non- aqueous solvent system • Any organic solution such as alcohol are used as a solvent. • They react with the sample and extract them during soaking. • Later the extract have to be separated using separatory funnel or rotary evaporator upon heating.

  14. Process Flow Diagram

  15. METHOD

  16. Solvent Extraction Equipments • Large Capacity Pump-mix Mixer-settler. • Industrial scale solvent extractor (frying roller, screw oil press, heating furnace, refining sets, winterization sets).

  17. Thank You

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