HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY Flashcards
• Technique used in analytical chemistry to separate, identify, and quantify specific components in a mixture
• Have high resolution and separation capacity
• Used in quantitative and qualitative analysis
High Performance Liquid Chromatography (HPLC)
High performance liquid Chromatography (HPLC) can only analyze mixtures dissolve in ___
Solvents
(basic principles)
High performance liquid Chromatography (HPLC) depends on the nature of stationary phase whether if it is an ____ chromatography or ____ chromatography
Absorption
Partition
2 commonly used type of HPLC
Absorption and Partition chromatography
Type of chromatography where separation is based on absorption
Absorption chromatography
Type of chromatography where separation is based on partition
Partition chromatography
(basic principle)
The separation principle of HPLC is based on the distribution of sample (analyte) between the ____ and ____
Mobile phase (eluent)
Stationary phase (packing material in column)
The time which is required for the compounds to be transported from the injection site to the detector unit is called ___
Retention time
Retention time the data for ____ and ___ of the compounds in the mixture
identification
quantification
(basic principle)
HPLC can separate compounds in the mixture through their interaction with ____, wherein the higher polarity, the slower it will move through the column
Polar stationary phase
Higher polarity = ?
Slower movement through column
(Basic principle)
Compounds are identified through the difference in their peak’s ___ time, with prior injected ____ standards
retention
reference
____ and related data from the detector helps calculate the concentration of each compound (or quantify specific compounds
Chromatogram
The more concentrated, the ?
stronger the signal
General components of HPLC
Solvent
Pump
Injector
HPLC column
Detector
PC (for data acquisition)
Waste container
Are a group of organic liquid used in HPLC
Solvents
Generates a flow of eluent from the solvent reservoir into the system
Pump
• liquid that dissolves the target compound
• polar and non polar mixture of solvent can be used
Mobile phase