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Index
»
QUALITY CONTROL 2 LEC
»
QC2- CHROMATOGRAPHY
»
Level 4
level: Level 4
Questions and Answers List
LC AND HPLC
level questions: Level 4
Question
Answer
STATIONARY PHASE: packing material MOBILE PHASE : liquid simple, versatile, high resolution separations
LIQUID CHROMATOGRAPHY (LC)
LIQUID CHROMATOGRAPHY (LC) can be performed using either of 2 METHODS:
1 Open column chromatography 2 HPLC
mobile phase is allowed to flow thru packed column under influence of gravity
Open column chromatography
– under high pressure - 1000 to 3000 psi required
(High-performance liquid chromatography) HPLC
ADSORPTION CHROMATOGRAPHY 1 MOST COMMONLY used adsorbents for Thin-layer chromatography
silica gel and alumina
reverse phase is most frequently used method in HPLC
Partition chromatography
HPLC Solvent reservoirs
1 Glass or stainless container 2 Solvent w/ highest purity 3 Contain the mobile phase 4 Mobile phase should be filtered, degassed using vacuum and sonication
Straight length of stainless tubing 5 to 25 cm in length 2 to 4.6 mm in diameter C8 (Octyl) and C18 (Octadecyl) modified adsorbents – most popular, almost 80% HPLC operations have been developed w/ these adsorbents THE HEART of HPLC separation process
Analytical Column
most popular, almost 80% HPLC operations have been developed w/ these adsorbents
C8 (Octyl) and C18 (Octadecyl) modified adsorbents
detectors used in HPLC: Optical detectors:
1 UV/VIS 2 IR/ raman 3 Optical activity 4 refractive index 5 Luminescent 6 Fluorescence
Mass Spectrometry detectors:
1 Fourier Transform Ion Cyclotron Resonance 2 Mass spectrometry (FT-MS)
Elemental Detectors:
1 AA/ Emission (Atomic absorption) 2 ICP-MS (Inductively coupled plasma mass spectrometry)
for oxidizable or reducible compound (catecholamines)
Electrochemical Detectors
Electrochemical Detectors
1 Conductivity 2 Coulometry 3 Polarography