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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

  • Kalaiselvi a/p Mohanraj SB09031

  • G.Sumithalaksmy

  • a/p Gunasegaran SB09063

  • Calvin Lee Weng Leong SB09017


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.


Introduction
INTRODUCTION metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.

  • 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.


CONT…. metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.

  • There is a need for efficient and economical approaches to the bioseparation problems.

  • Extraction using aqueous two-phase is one such possibility.


Principle of extraction
Principle of Extraction metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.

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%)


Principle of extraction1

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


  • Nucleic acid extraction methods: metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.

    • 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


  • The metabolic compounds, protein & nucleic acid. Include the principle of extraction, common method &equipment used in each extraction process.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.


Influencing factors of target compound s partition coefficient in aqueous two phase system
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


Equipments used
Equipments Used Coefficient in Aqueous

  • Mixer settler unit

  • Centrifuge tube

  • Centrifuge machine

  • Vortex


PRINCIPLE OF CONVENTIONAL EXTRACTION SYSTEM Coefficient in Aqueous

  • 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.


Aqueous solvent system Coefficient in Aqueous

  • 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.


Non- aqueous solvent system Coefficient in Aqueous

  • 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.


Process Flow Diagram Coefficient in Aqueous


METHOD Coefficient in Aqueous


Solvent extraction equipments
Solvent Extraction Equipments Coefficient in Aqueous

  • Large Capacity Pump-mix Mixer-settler.

  • Industrial scale solvent extractor (frying roller, screw oil press, heating furnace, refining sets, winterization sets).


Thank You Coefficient in Aqueous


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