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    Chapter 26 - Gas Chromatography1

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    PARIK
    The document discusses gas chromatography (GC) and its use in analyzing pesticide residues in tea samples. Key points: 1) Samples were placed in sealed vials and underwent headspace solid phase microextraction (HS-SPME) to extract compounds, which were then thermally desorbed into the GC for separation and analysis. 2) GC/TOF MS chromatograms show peaks for pesticides in spiked tea samples but not blank samples, demonstrating the technique can detect pesticide residues. 3) Detection limits for some pesticides like malathion and heptachlor were in the low μg/kg range, below regulatory limits, showing the method is suitable for pesticide residue analysis

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    Chapter 26 - Gas Chromatography1

    Uploaded by

    PARIK
    72 views19 pages
    The document discusses gas chromatography (GC) and its use in analyzing pesticide residues in tea samples. Key points: 1) Samples were placed in sealed vials and underwent headspace solid phase microextraction (HS-SPME) to extract compounds, which were then thermally desorbed into the GC for separation and analysis. 2) GC/TOF MS chromatograms show peaks for pesticides in spiked tea samples but not blank samples, demonstrating the technique can detect pesticide residues. 3) Detection limits for some pesticides like malathion and heptachlor were in the low μg/kg range, below regulatory limits, showing the method is suitable for pesticide residue analysis

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    The document discusses gas chromatography (GC) and its use in analyzing pesticide residues in tea samples. Key points: 1) Samples were placed in sealed vials and underwent headspace solid phase microextraction (HS-SPME) to extract compounds, which were then thermally desorbed into the GC for separation and analysis. 2) GC/TOF MS chromatograms show peaks for pesticides in spiked tea samples but not blank samples, demonstrating the technique can detect pesticide residues. 3) Detection limits for some pesticides like malathion and heptachlor were in the low μg/kg range, below regulatory limits, showing the method is suitable for pesticide residue analysis

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    72 views19 pages

    Chapter 26 - Gas Chromatography1

    Uploaded by

    PARIK
    The document discusses gas chromatography (GC) and its use in analyzing pesticide residues in tea samples. Key points: 1) Samples were placed in sealed vials and underwent headspace solid phase microextraction (HS-SPME) to extract compounds, which were then thermally desorbed into the GC for separation and analysis. 2) GC/TOF MS chromatograms show peaks for pesticides in spiked tea samples but not blank samples, demonstrating the technique can detect pesticide residues. 3) Detection limits for some pesticides like malathion and heptachlor were in the low μg/kg range, below regulatory limits, showing the method is suitable for pesticide residue analysis

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 19

    Gas Chromatography

    Let’s begin with an example problem: SPME head space analysis of


    pesticides in tea and follow-up analysis by high speed GC.

    Samples in 10mL sealed glass vials were placed in the MPS-2 autosampler for HS-
    SPME extraction. Extracted compounds were thermally desorbed from the fiber
    after its insertion into the hot (270 ◦C) splitless GC injection port. Desorption
    time was 2min.
    Anal Chim. Acta 611 (2008) 163
    Gas Chromatography
    Conventional HS-SPME

    Fig. 3 – Comparison of conventional (a) and SPME-based (b) approach in analysis of


    pesticide residues in tea samples. GC/TOF MS chromatograms of blank (non-spiked)
    black (I) and fruit (II) teas; pesticide standard solution (200g kg−1) (III).

    Malathion Heptachlor Anal Chim. Acta 611 (2008) 163


    Gas Chromatography
    Detection Figures of Merit (36 pesticides tested)

    Pesticide MRL Linearity Dynamic LOQ RSD (%)


    (μg/kg) (r2) Range (μg/kg) 50 μg/kg
    (μg/kg) S/N = 12 N=5

    HS-SPME

    Malathion 500 0.9838 50-200 26 13

    Heptaclor 20 0.9870 10-200 8 9

    Ethyl Very time


    acetate consuming !
    extraction
    Malathion 500 0.9977 10-200 8 11

    Heptaclor 20 0.9983 1.25-200 1 5

    MRL = maxiumum residue limit


    Gas Chromatography - Instrument
    A gas chromatograph (GC) is an analytical instrument that measures the content of
    various volatile components in a sample. The analysis performed by a gas
    chromatograph is called gas chromatography.

    Principle of gas chromatography: The sample solution injected into the instrument
    enters a gas stream which transports the sample into a separation tube known as the
    "column." (Helium or nitrogen is used as the so-called carrier gas.) The various
    components are separated inside the column. The detector measures the quantity of
    the components that exit the column.

    Internet image
    Gas Chromatography
    Column separation (gas-liquid, gas-solid) used for separating and
    analyzing compounds that can be vaporized without decomposition.

    Higher b.p Lower b.p

    Separations generally based on


    differences in boiling points!
    Conditions

    • Carrier gas (mobile phase) does NOTHING in GC but transport the


    compounds. Not involved in separation mechanism (H2 and He common).

    • Injection volume (0.1 – 10 μL generally). Temperature of injector is 50 oC


    greater than least volatile (highest boiling point compound). All
    compounds must be vaporized before transport onto column.

    • Fixed temperature separation – average boiling point of all analytes is a


    good starting point.

    • Carrier gas is often dried by passage over molecular sieves as they strongly
    retain water. Activated by heating to 300 oC in vacuum.

    • Gaseous mobile phase carries gaseous compounds (analytes) through a


    long column with a stationary phase.
    Thermodynamics of Separations
    The application of thermodynamics to a chromatographic separations
    explains how the distribution coefficient (which itself determines the
    magnitude of retention) is controlled by the standard energy of
    distribution and the absolute temperature.

    CStat
    KD 
    CMobile
    Amobile AStationary

    GD
      ln K D
    RT
    The Chromatogram
    Average velocity
    of moving
    analyte zone

    tr  tm kb (last )
    k 
    '
    
    tm k a ( first )
    Capacity Factor (2-10) Selectivity Factor (>1)
    Plate Height – Equilibration Zones

    Distillation theory

    Height equivalent to a theoretical plate (H)

    L(cm)
    N ( platenumbe r ) 
    H (cm / plate )
    2
     tr   t r2 
    N  16  N  5.55 2 
    W   W1/ 2 
    Why Do Bands Separate and Broaden with
    Time on Column?

    Longer on the column, longer the retention time (tr) and the broader the peaks.
    Why Does Peak Width Matter?

    Although the selectivity factor, α, describes the separation of band centers, it


    does not take into account peak widths. Another measure of how well species
    have been separated is provided by measurement of the resolution (R). The
    resolution of two species, A and B, is defined as:

    Rs 

    2 t rb  t ra 
    wb  wa
    w = baseline peak width of two neighboring peaks.

    Baseline resolution, R > 1.5


    Rate Theory of Chromatography

    A more realistic description of the processes at work inside a column takes account
    of the time taken for the solute to equilibrate between the stationary and mobile
    phase (unlike the plate model, which assumes that equilibration is infinitely fast).
    The resulting band shape of a chromatographic peak is therefore affected by the
    rate of elution. It is also affected by the different paths available to solute molecules
    as they travel between particles of stationary phase. If we consider the various
    mechanisms which contribute to band broadening, we arrive at the Van Deemter
    equation for plate height;

    B
    H  A   Cu
    u
    where u is the average velocity of the mobile phase. A, B, and C are factors which
    contribute to band broadening.
    Rate Theory of Chromatography

    Van Deemter plot -


    A plot of plate height (H) vs. average linear velocity (u) of mobile phase.
    Rate Theory of Chromatography
    A - Eddy diffusion
    The mobile phase moves through the column which is packed with stationary phase.
    Solute molecules will take different paths through the stationary phase at random. This
    will cause broadening of the solute band, because different paths are of different
    lengths.

    B - Longitudinal diffusion
    The concentration of analyte is less at the edges of the band than at the center. Analyte
    diffuses out from the center to the edges. This causes band broadening. If the velocity of
    the mobile phase is high then the analyte spends less time on the column, which
    decreases the effects of longitudinal diffusion.

    C - Resistance to mass transfer


    The analyte takes a certain amount of time to equilibrate between the stationary and
    mobile phase. If the velocity of the mobile phase is high, and the analyte has a strong
    affinity for the stationary phase, then the analyte in the mobile phase will move ahead of
    the analyte in the stationary phase. The band of analyte is broadened. The higher the
    velocity of mobile phase, the worse the broadening becomes.
    Efficiency of Separation (plate number)

    H = L/N Small plate height = narrow peaks = better separation

    Multiple Flow Path Term


    One wants a small value for H! Plate numbers 100s-1000s for conventional
    HPLC. Can be 106 for some high efficiency separations!!!!
    Zone Broadening Terms

    A term = Eddy Diffusion

    Bterm = Longitudinal Diffusion

    C term = Stationary Phase


    Mass Transport
    Why Do We Care About Flow Rate?
    N (efficiency) = L/H N α Rs

    Van Deemter Eq.

    Flow Velocity (cm/s) = Flow Rate (cm3/s)/Cross sectional area (cm2)


    Rate Theory of Chromatography

    Internet image
    Rate Theory of Chromatography

    H is plate height
    λ is particle shape (with regard to the packing)
    dp is particle diameter
    γ, ω, and R are constants (function of packing and k’)

    Dm is the diffusion coefficient of the mobile phase


    dc is the capillary diameter
    df is the film thickness
    Ds is the diffusion coefficient of the stationary phase.
    u is the linear velocity
    Goal: To achieve the fastest separation with the highest plate
    number and resolution!!!

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