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LC/MS Techniques in Analytical Chemistry

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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vinaynaithani
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110 views22 pages

LC/MS Techniques in Analytical Chemistry

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.

Uploaded by

vinaynaithani
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd

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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