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

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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 LeongSB09017.

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

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  • Group Members:

  • Rubini Devi a/p SelvarajooSB09005

  • Kalaiselvi a/p MohanrajSB09031

  • G.Sumithalaksmy

  • a/p GunasegaranSB09063

  • Calvin Lee Weng LeongSB09017

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.


  • 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.


  • 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

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 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)


polymer-low molecular weight compound(salting-out effect)

PEG-Ammonium Sulfate

  • 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

  • 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.

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

  • Mixer settler unit

  • Centrifuge tube

  • Centrifuge machine

  • Vortex


  • 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

  • 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

  • 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


Solvent Extraction Equipments

  • Large Capacity Pump-mix Mixer-settler.

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

Thank You

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