Presented By

S.SUBALAKSHMI
II-BIOCHEMISTRY
14UBCH039

Gas
Chromatography
Function
Components
Common uses
Chromatographic resolution
Sensitivity

Function
Separation of volatile organic compounds
Volatile when heated, VOCs undergo a

phase transition into intact gas-phase species

Separation occurs as a result of unique
equilibria established between the solutes and
the stationary phase (the GC column)
An inert carrier gas carries the solutes
through the column

Polar vs. nonpolar

Separation is based on the vapor pressure

and polarity of the components.

Within a homologous series (alkanes, alcohol,
olefins, fatty acids) retention time increases
with chain length (or molecular weight)
Polar columns retain polar compounds to a
greater extent than non-polar
C18 saturated vs. C18 saturated methyl ester

Detectors
Flame Ionization Detectors (FID)
Electron Capture Detectors (ECD)
Electron impact/chemical ionization (EI/CI)

Mass spectrometry

Parts of a Mass Spec

Sample introduction
Source (ion formation)
Mass analyzer (ion sep.) - high vac
Detector (electron multiplier tube)

Sample
Introduction/Sources
Volatiles
Probe/electron impact (EI),Chemical ionization

(CI)
GC/EI,CI
Involatiles
Direct infusion/electrospray (ESI)
HPLC/ESI
Matrix Assisted Laser Adsorption (MALDI)
Elemental mass spec
Inductively coupled plasma (ICP)
Secondary Ion Mass Spectrometry (SIMS)
surfaces

Mass Analyzers
Low resolution
Quadrupole
Ion trap
High resolution
TOF time of flight
Sector instruments (magnet)
Ultra high resolution
ICR ion cyclotron resonance

Mass accuracy
Mass Error = (5 ppm)(201.1001)/10 6 =

0.0010 amu
201.0991 to 201.1011 (only 1 possibility)
Sector instruments, TOF mass analyzers
How many possibilities with MA = 50 ppm?
with 100 ppm?

Exact Mass
Determination

Need Mass Spectrometer with a high mass

accuracy 5 ppm (sector or TOF)

C9H15NO4, FM 201.1001 (mono-isotopic)
Mass accuracy = {(Mass Error)/FM}*106
Mass Error = (5 ppm)(201.1001)/10 6 =

0.0010 amu