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    Home > Biochemistry News > Biotechnology News > Ion exchange resin layering separates mixed amino acids.

    Ion exchange resin layering separates mixed amino acids.

    • Last Update: 2020-10-26
    • Source: Internet
    • Author: User
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    related topics . Objective: To understand
    working principle and operating techniques of ion exchange resin layering.
    2. Learn to separate the amino acids of the
    by ion exchange resin
    .ion exchange resin
    a synthetic polymer, insoluble in water, can absorb water expansion. Polymer molecules consist of ionization-ionization of the sex group and non-polar resins. The ions on the polar group can exchange with the ions in the solution, and the non-polar resin itself remains the same. Usually the ion exchange resin is divided into strong acids (-R-S03H), weak (-COOH), strong alkalis (-N-R:) and weak alkalis (-NH2-NHR-NR2) according to the group.
    ion exchange resin to separate small molecules such as amino acids, adenosine, adenosines, etc. is more ideal. However, it is not appropriate
    biomass
    such as proteins, because they cannot spread to the chain structure of the resin. Therefore, such as the separation of biological large son, optional polysaccharide polymers such as cellulose, glucosaccharide as the carrier of ion exchange agent.
    this experiment used sulfonate cation exchange resin to isolate a mixture of acidic amino acids (tiandongine) and neutral amino acids (alanine) alkaline amino acids (lysine). Under certain pH conditions, they are dissodiated to varying degrees and can be separated by changing the pH or ion strength of the dehydration.Experimental Materials
    1. Experimental equipment
    -layering column (1.6X20cm); constant current pump; gradient mixer;
    test tube
    and test tube rack; ultraviolet
    phosphorescometer
    , sulfonate cation exchange resin (Dowex 50)
    2. Experimental

    (1)2mol/L HCl
    (2) 2mol/L NaOH
    (3) 0.1mOl/L HCl
    (4) 0.1mol/L NaOH

    (5) pH4.2 citric acid buffer: 0.lmol/L citric acid 54m1 plus 0.1mol/L sodium citrate 46ml
    (6) pH5 acetic acid buffer: 0.2mol/L NaAc 7 0ml plus 0.2mol/L HAc 30ml
    (7) 0.2% neutral tritone solution: 0.2g tritone plus 100ml acetone
    (8) amino acid mixture: propylene Arionine, tiandonine, lysine each 10m1 plus 0.1mol/L HCl 3m(Experimental Method)
    1. Treatment of resin
    100ml
    Beast
    with about 10g resin, plus 25ml. 12mo1/L HCl stirs 2h, discards the acid and fills the washing resin with distilled water to neutral. Add 25ml 12mol/L NaOH to the above resin and stir 2h, discard the lye and wash with distilled water to neutral. Suspend the resin in a 50 ml pH4.2 citric acid buffer.
    2. Load column
    take the diameter of 0.8cm to 1.2cm, length 10cm to 12cm of the laminate column, bottom pad glass cotton or sponge round pad, from the top into the treated resin suspension, close the column outlet, after the resin subsides, release excess solution, after adding some resin, to the resin deposit to 8cm to 10cm height can be. Continue to add pH4.2 citric acid buffer to the top of the column to wash, so that the outflow pH is 4.2, close the column outlet, keep the liquid surface about 1cm above the resin surface.
    3. Dosing, wasting and wasting
    open the yamagun to allow the buffer to flow out and close the outlet when the liquid surface is almost flat on the resin surface (do not dry the resin surface ). Use a long dropper to carefully add 15 thymino acid mixtures directly to the top of the resin, opening the outlet so that it slowly flows into the column.
    When the liquid surface is just flat resin surface, add 0.1mol/L HCl 3 ml, at 10 drops / min to 12 drops / min flow rate wash away, collect semen, 20 drops per tube, tube-by-tube storage. When the HCl liquid surface just flat resin surface, with 1m1 pH4.2 citric acid buffer rinsed the column wall once, and then with 2 ml pH4.2 citric acid buffer washed away, to maintain the flow rate of 10 drops / min to 12 drops / min and pay attention not to dry the resin surface.
    In the process of collecting the sequestration, step by tubetritone test amino acid enchantment, the method is: in each tube desalination add 10 drops of pH5 acetic acid buffer and 10 drops of neutral The depth of the color can represent the concentration of the washed-out amino acids, which can be measured by compared color.
    .
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