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    Liquid Chromatography & Mass Spectrometry: BCH 4028 Instrumental Analytical Chemistry

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    vinaynaithani
    This document discusses liquid chromatography-mass spectrometry (LC-MS), providing information on sample types, selectivity, sensitivity, interfacing LC with MS, ionization techniques including electrospray and atmospheric pressure chemical ionization, scan vs selective ion monitoring acquisition modes, collision-induced dissociation, adapting LC methods for MS, and applications for molecular weight determination, pharmaceutical analysis, and environmental analysis. Key aspects covered are the high resolving power and specificity of LC-MS for analyzing specific sample components.

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    Liquid Chromatography & Mass Spectrometry: BCH 4028 Instrumental Analytical Chemistry

    Uploaded by

    vinaynaithani
    84 views22 pages
    This document discusses liquid chromatography-mass spectrometry (LC-MS), providing information on sample types, selectivity, sensitivity, interfacing LC with MS, ionization techniques including electrospray and atmospheric pressure chemical ionization, scan vs selective ion monitoring acquisition modes, collision-induced dissociation, adapting LC methods for MS, and applications for molecular weight determination, pharmaceutical analysis, and environmental analysis. Key aspects covered are the high resolving power and specificity of LC-MS for analyzing specific sample components.

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    LCMS

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    This document discusses liquid chromatography-mass spectrometry (LC-MS), providing information on sample types, selectivity, sensitivity, interfacing LC with MS, ionization techniques including electrospray and atmospheric pressure chemical ionization, scan vs selective ion monitoring acquisition modes, collision-induced dissociation, adapting LC methods for MS, and applications for molecular weight determination, pharmaceutical analysis, and environmental analysis. Key aspects covered are the high resolving power and specificity of LC-MS for analyzing specific sample components.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    84 views22 pages

    Liquid Chromatography & Mass Spectrometry: BCH 4028 Instrumental Analytical Chemistry

    Uploaded by

    vinaynaithani
    This document discusses liquid chromatography-mass spectrometry (LC-MS), providing information on sample types, selectivity, sensitivity, interfacing LC with MS, ionization techniques including electrospray and atmospheric pressure chemical ionization, scan vs selective ion monitoring acquisition modes, collision-induced dissociation, adapting LC methods for MS, and applications for molecular weight determination, pharmaceutical analysis, and environmental analysis. Key aspects covered are the high resolving power and specificity of LC-MS for analyzing specific sample components.

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    of 22

    BCH 4028

    Instrumental Analytical Chemistry

    Liquid chromatography
    & Mass spectrometry

    These notes are


    from Agilent
    Literature
    LC/MS
    LC MS
    provide information about the
    high molecular high detection
    resolving weight, structure,
    power specificity
    identity & quantity of
    specific sample components
    Sample types

     sample types range from small


    pharmaceutical compounds to large proteins
     suitable for the
    analysis of large,
    polar ionic, thermally
    unstable & involatile
    compounds
    Sample types
    Selectivity & Sensitivity

    High
    selectively
    Detect High
    compounds sensitivity
    of interest
    in a complex Detect only
    matrix those masses
    characteristic
    of the
    compounds
    being
    monitored
    Interfacing LC & MS

    LC MS

    Pressure High pressure to Vacuum


    separate a liq. phase Limited gas
    Produces a high gas load load
    Flow eq. to 1 L/min gas 1 mL/min
    Temp. Near ambient temp. Elevated temp.
    Mass range No Yes
    limitation

    Buffer Inorganic buffers Prefers volatile


    buffers
    API-Electrospray

    Analyze samples that become


    multiply charged: proteins, peptides &
    oligonucleotides
    singly charged: benzodiazepines & sulfated
    conjugates

    Process:
    nebulization Ion
    & charging desolvation
    evaporation
    http://www.chm.bris.ac.uk/ms/theory/esi-ionisation.html
    API-Electrospray

    charged droplets shrinks droplets


    carries away uncharged
    species
    Atmospheric pressure chemical ionization: APCI

    Applicable to a wide range of polar &


    nonpolar analytes that have moderate
    molecular weights

    Process:
    nebulization &
    desolvation ionization
    nebulization occurs in a by corona
    hot(typically 250 oC–400 oC) discharge
    vaporizer chamber to
    evaporate droplets
    APCI
    Scan vs. SIM

    Scan
    – detects signals over a mass range
     max. qualitative info.

    Selective Ion Monitoring


    – only monitor a few m/z ratios
     max.
    quantitative info.
    Scan vs. SIM
    CID

    MS/MS is accomplished by a process


    called collision-induced dissociation in
    which ions break apart as a result of
    collisions with other molecules

    EI can produce CID by using higher


    voltages on the electrode
    CID

    single ion can be selected


    and fragmented
    Adapting LC methods

     Orthogonal spray ion source is very


    tolerant of non-volatile components in
    samples or mobile phase
     The flow is directed orthogonal to the lens
    axis & ions are focused into the mass filter.
     Orthogonal spray orientation & high-
    capacity drying gas system improve
    instrument performance &  maintenance
    requirements.
    Adapting LC
    methods: orthogonal
    spray
    Applications

     Molecular weight determination


     Combinatorial chemistry
     Pharmaceutical applications
     Biochemical applications

    Clinical applications
     Food applications
     Environmental applications
    Molecular weight determination Applications

    Examples

    C-terminus: threonine vs. threonine amide


    API-ES

    Determining molecular weight of green


    fluorescent protein by API-ES

    Between m/z 1000 and 1300 there are


    6 peaks. Two of these adjacent peaks
    are at 1118.85 and 1167.55.
    What is the parent ion mass?
    Pharmaceutical applications Applications

    peaks unresolved
    Environmental applications Applications

    have 1 ring and


    differ by 1 Cl

    has 2 Cl and
    a 2nd ring
    attached to
    the first by
    O
    Further
    reading
    http://www.chem.agilent.com/scripts/Li
    teratureresults.asp?iprodinfotype=4&ip
    rodgroup=8

    Videos
    http://www.chem.agilent.com/Scripts/
    Generic.ASP?lPage=8683&indcol=N&pro
    dcol=Y

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