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    PG 504 Sec 8

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    selenaalizx

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    © All Rights Reserved
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    PG 504 Sec 8

    Uploaded by

    selenaalizx
    4 views11 pages

    Original Title

    PG 504 sec 8

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    4 views11 pages

    PG 504 Sec 8

    Uploaded by

    selenaalizx

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
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    of 11

    Sec 8

    Advanced Column
    chromatography
    (GC)
    Principle
     Gas chromatography is a process of separating volatile substances
    of a mixture by using gaseous mobile phase.”

     A sample being vaporized and injected onto the chromatographic


    column then transported through the column by the flow of inert,
    gaseous mobile phase.

     Two major types:

    • Gas-solid chromatography; GSC (adsorption mechanism).

    • Gas-liquid chromatography; GLC (partition mechanism).


    GC
    Components:
    Instrumentation
     Gas cylinder/tank is fitted with a flow controller to ensure a constant flow rate
    of mobile phase to the column.
    - Carrier gas may be: He (common), N2, H2, Argon
     Sample injection port

    - Micro syringe is used for sampling or Automatic injectors.


     Column (inside oven)
    - The sample gets vaporized and flows into column by the carrier gas.
     Detector
    - Thermal conductivity (TCD)
    - Electron capture detector (ECD)
    - Flame ionization detector (FID)
    Derivetization in GC
    Definition:

    Chemical modification produce new compound which has properties that


    are suitable for analysis in GC.

    Importance:

    1-Indirect way to derive the compounds to be volatile and stable.

    2-Increase detectability & sensitivity specially in case of Electron


    capture detector(ECD).
    Types of derivetization
    1- Silylation: Si(CH3)3
    It is the most prevalent method used to increases volatility of
    compounds.

    2- Alkylation:
    Used to protect certain function groups in compounds then further
    derivetization can be made.

    3- Acylation: Acyl-halide
    Commonly used to add fluorinated groups (ECD).
    Chromatogram
    net result of separation might be :

    1) Spots in case of TLC and PC.

    2) Zones or Bands in case of classical columns.

    3) peaks of chart in case of advanced devises such as


    GC and HPLC.

    The series of (spots, zones, bands or peaks) appear on


    the (TLC, PC, Column or chart) after
    (development/Elution) in case of (Planner or
    columner) chromatography.
    Experiment 4
    Using dragendorffꞌs reagent as a specific reagent
    for detection of colorless alkaloids

    Atropine, brucine & papaverine.

    St. phase: silica gel (TLC)

    M. phase: CHCl3:EtOH(85%:15%)
    1- TLC preparation
    2- Sample application (spotting)

    Atropine

    Sample brucine

    Papaverine
    2- Development.

    3- Detection(Visualization) of the separated spots.


    5- Calculation of Rf value :
    Rf = distance traveled by the sample spot /
    distance traveled by m. phase front

    6 - Comment

    Unk A B P

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