View
236
Download
2
Embed Size (px)
Citation preview
Chromatography
Gas
GSC GLC
Liquid
LSC LLC IEC GPC AC
Gas Chromatography
Mobile phase :gas (usually He, N2, H2)
Stationary phase :non-volatile liquid or solid particles
Analytes for GCHave a significant vapor pressureThermal stable
ex : gas, most non-ionized small and medium (C25) organic molecules, many organometallic compounds.
Not for organic and inorganic salts, macromolecules
樣品形式: solid, liquid, gas
injectionport
exit
detector
septum
carrier gasin
column oven
column
computer orrecorder
detectoroven
injectoroven
Schematic diagram of a gas chromatography
Carrier gas
no reactivenon flammablenon-toxic
N2, He, H2
H
μAB
C
CBAH
N2→packed columnHe, H2→open tube column
N2
He
H2
Pla
te h
eigh
t (m
m)
Linear velocity (cm/s)
H2 and He both give optimal resolution at higher flow rates than N2
Column
flow0.1 ~ 0.53 mminner diameter
stationary phase(0.1 ~ 5μm thick)
outer wall of columnOpen tubular columns
Packed columncontain a fine solid support coated with a nonvolatile liquid stationary phase; or the solid itself may be the stationary phase.
Packed column Open tubular column
I.D. 2 ~ 6 mm 0.2 ~ 0.75 mm
L 0.5 ~ 5 m 15 ~ 100 m
Open tubular columns
Compared with packed columns, open tubular
columns offer :
1. higher resolution
2. shorter analysis time
3. greater sensitivity
4. lower sample capacity
wall-coatedopen tubular column
(WCOT)FSOT: fused-silica, OTC
support-coatedopen tubular column
(SCOT)
porous-layeropen tubular column
(PLOT)
Decreasing thickness of stationary phase leads to1. decreased plate height2. decreased retention time3. decreased capacity for analyte
GSC
separation based on surface adsorption
Absorbent :Molecular sieves : for H2, O2, N2, CH4, CO (PLOT)
Charcoal : for permanent gas C1 ~C4, stack gas (NOx, SOx)
Silica gel, alumina (PLOT)
Porous polymer
GLC
a solid support coated with a liquid stationary phase
Solid support for GLC :Diatomaceous earths (diatom: single-cell algae)
graphite carbon, charcoal
Teflon
Liquid phase for GLC :Chemical stable and unreactive toward analyte
Involatile and thermal stable
OHSiCl Si
R
R'
R"
O SiSiR
R'R"
silanol
Separation mechanism
(1) selective solute-S.P. interaction
(2) difference in vapor pressure of solute
S.P. 分類non-polar phase
polar phase
special phase
chiral phase
What is the chiral molecule?
對掌性異構物 (enantiomers):具有相同的物性及化性。Ex: boiling point 、 melting point 、 density 、 refractive indexes 、 ionization 、 reaction properties 。
Mirror
C
OH
CH2OHHOC
H
C
HHO
COHHOH2C
Chiral compound 的重要性
(1) 可運用於製藥、農藥、食物及飲料。(2) 瞭解具有生物活性分子的生物合成及代謝。(3) 使用的分離技術: HPLC 、 GC 、 CE …etc 。
R formS form
鎮靜效果畸胎原 Different biological effect
譬如:
N
O
ONH
O
O
*
thalidomide
莎痢竇邁
對掌固定相 Chiral Stationary Phase(CSP)1.Dalgiesh 1952 three-point interaction2. 暫時性的非對映異構錯合物
transient diastereomeric complex
流動沖堤方向R form
S form
Parameters influence on selectivity
The nature of the S.P.
The conc. of S.P.
Column T • isothermal temp. control
• programming temp. control
The choice & pretreatment of the solid support
Temperature and pressure programmingD
etec
tor
resp
onse C6 C7
C9air C8
Low T
C6 C7 C9
airC8 C10 C11 C12 High T
Det
ecto
r re
spon
se
C6 C7 C9
airC8 C10 C12C11 C13
time
T
Det
ecto
r re
spon
se
Retention index (I)
)(log)(log
)(log)(log)(100
''
''
ntNt
ntunknowntnNnI
rr
rr
n : # of C atoms in the smaller alkane N : # of C atoms in the larger alkanetr(n) : adjusted retention time of smaller alkane tr(N) : adjusted retention time of larger alkanetr(unknown) : adjusted retention time of unknown
Retention index relates the retention time of a solute to the retention times of linear alkanes.
C8
Time (min)3.50.25
Det
ecto
rR
espo
nse C7
C9
xair
5.6 7.46.3
Kovats index for linear alkanes equals 100 times the
number of carbon atoms
0.25)-6.5log(0.25)-4.7( log0.25)-6.5log(0.25)-(6.3 log
8)-(9 8100 I
I = 842
Injection conditions for split, splitless, and on-column injection
Syringe for solid-phase microextraction.The fused silica fiber is withdrawn inside the steel needle after sample collection and when the syringe is used to pierce a septum.
Sampling by solid-phase microextraction and desorption of analyte from the coated fiber into a gas chromatography.
Purge and trap apparatus for extracting volatile substances from a liquid or solid by flowing gas.
Detectors
Thermal Conductivity detector
gas
e-
gas
e-
high thermal conductivitymain component entering detector
analyte appears, T.C., Temp , Wire Resistance , detector current “read-out”
Thermal conductivity detector :104 linear response range
H2 and He give lowest detection limit
sensitivity increase with
increasing filament current
decreasing flow rate
lower detector block temperature
Flame ionization detector (FID)
column effluent
H2
air
cathode(collects CHO+ ions)anode
Carbon atoms (except carbonyl and carboxyl carbons) produce CH radicals, witch are though to produce CHO+ ions in the flame :
CH + O → CHO+ + e-
Flame ionization detector :H2 give best detection limit
Signal proportional to number of susceptible carbon atoms
100-fold better detection than thermal conductivity
107 linear response range
Insensitive to non-hydrocarbons
Electron capture detector (ECD)
electrodes
Radioactive-emitter
Ni63 +
+
insulator
-
-
- + gas - + gas+ + e- (standing current)e- + solute solute- (e- capture)solute- + gas+ solute + gas
time
Cur
rent
(I)
standing current
Electron capture detector :Non-destructive
non-linear
selective to e- capturing solutes
detection limit 5 fg
Other detectorsElectron capture : halogens, conjugated C=O, -C≡N, -NO2
nitrogen-phosphorus : highlights P, N
Photoionization : aromatics, unsaturated compounds
Sulfur chemiluminescence : S
Nitrogen chemiluminescence : N
Atomic emission : most elements (selected individually)
Mass spectrometer : most analytes
Infrared spectrometer : most analytes.
Name Type Selective For : Typical Min. Detectible Level (S/N=2)
Linear Dynamic Range
FID SelectiveMaterials that ionize in air/H2 flame
5 pg C/sec 107
TCD UniversalAnything with thermal cond. Different from carrier
400 pg/ml carrier 106
ECD Selective Gas-phase electrophores0.1 pg Cl/sec(varies w/structure)
104
PID Selective Compounds ionized by UV 2 pg C/sec 107
Thermionic Selective N, P, heteroatoms 0.4 pg N/sec ; 0.2 pg P/sec 104
ELCD Selective Halogens, N, S0.5 pg Cl/sec ; 2 pg S/sec4 pg N/sec
106 ; 104
104
FPD Selective P, S 20 pg S/sec ; 0.9 pg P/sec 103 ; 104
FTIR Universal Molecular vibrations 1000 pg of strong absorber 103
MSD Universal Tunable for any species 10 pg to 10 ng (depending on SIM vs. scan) 105
AED Universal Tunable for any species 0.1 – 20 pg/sec(depending on element) 104