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Separation
mechanism
Type of
stationary
phase
Fasa diam
(adsorpsi fisika)
Sistem partisi
Liquid-bonded
phase
Fasa diam
(ikatan kimia)
Fasa
diam
(polar)
Triethylene
glycol
Water
Normal phase
Fasa
gerak
(non
polar)
Hexane
Propyl ether
Fasa diam
(non polar)
Hydrocarbon
Fasa gerak
(polar)
Water
Methanol
Acetonitrile
THF
Reversed
phase
Pharmaceutical
Biochemicals
Food products
Industrial chemicals
Pollutants
Forensic chemistry
Clinical medicine
Lebih dari
menggunakan reversed phase, bonded phase
(octyl or octyldecyl-siloxane) packings dan fasa
geraknya antara lain metanol, acetonitrile, atau
THF
Adsorpsi (liquid-solid)
chromatography
Fasa diam berupa permukaan padatan polar
(silika dan alumina)
Silika lebih dipilih untuk kebanyakan aplikasi
karena kapasitas sampel yang lebih besar
Penggunaan kedua fasa diam ini membuat
waktu retensi lebih lama jika polaritas analit
meningkat
Fasa gerak adalah pelarut organik atau
campuran pelarut organik
komposisi
berpengaruh pada koefisien distribusi
Affinity chromatography
(Kromatografi Afinitas)
Involves covalently bonding a
reagent, called an affinity ligand, to a
solid support
Typical affinity ligands are antibodies,
enzyme inhibitors, or other
molecules that reversibly and
selectively bind to analyte molecules
in the sample
The retained analytes can be eluted
by changing the mobile phase
Mobile phase
Two distinct roles:
First it must support the strong binding of
the analyte molecules to the ligand
Second, once the undesired species are
removed, the mobile phase must weaken or
eliminate the analyte-ligand interaction so
that the analyte can be eluted
(often changes in pH or ionic strength are
used to change the elution condition during
the two stages of the process)
Stationary phase
A solid such as agarose or a porous
glass bead to which the affinity
ligand is immobilized
Lanjutan