HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

University of Babylon / College of Pharmacy
Department of Pharmaceutical Chemistry/Analytical Chemistry
Lecture number - 11 28/12/2018
STRUMENTAL METHOD:
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
OR HIGH-PRESSURE LIQUID CHROMATOGRAPHY
• Is a chromatogrphic technique that can separate a mixture of compounds, and is used in biochemistry and analytical chemistry to identify , quantify and purify the individual components of the mixture.

• HPLC utilizes different types of stationary phase [ typically , hydrophobic saturated carbon chains ] .

• A pump that movse the mobile phase(s) and analyte through the column.
• A detector that provides a characteristic retention time for the analyte , the detector may also provide other characteristic information ( i.e. UV / VIS spectroscopic data for analyte if so equipped).
• Analyte retention time varies depending on the strength of its interactions with the stationary phase the ratio /composition of solvent (s) used and the flow rate of the mobile phase.

• With HPLC , a pump (rather than gravity).

• Provides the higher pressure required to propel the mobile phase and analyte through the densely packed column . the increased density arises from smaller particle sizes.

This allows for a better separation on columns of shorter length when compared to ordinary column chromatography.
Thank you
1

University of Babylon / College of Pharmacy
Department of Pharmaceutical Chemistry/Analytical Chemistry
Lecture number - 11 28/12/2018
STRUMENTAL METHOD:
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
OR HIGH-PRESSURE LIQUID CHROMATOGRAPHY
• Is a chromatogrphic technique that can separate a mixture of compounds, and is used in biochemistry and analytical chemistry to identify , quantify and purify the individual components of the mixture.

• HPLC utilizes different types of stationary phase [ typically , hydrophobic saturated carbon chains ] .

• A pump that movse the mobile phase(s) and analyte through the column.
• A detector that provides a characteristic retention time for the analyte , the detector may also provide other characteristic information ( i.e. UV / VIS spectroscopic data for analyte if so equipped).
• Analyte retention time varies depending on the strength of its interactions with the stationary phase the ratio /composition of solvent (s) used and the flow rate of the mobile phase.

• With HPLC , a pump (rather than gravity).

• Provides the higher pressure required to propel the mobile phase and analyte through the densely packed column . the increased density arises from smaller particle sizes.

This allows for a better separation on columns of shorter length when compared to ordinary column chromatography.
Thank you
1

University of Babylon / College of Pharmacy
Department of Pharmaceutical Chemistry/Analytical Chemistry
Lecture number - 11 28/12/2018
STRUMENTAL METHOD:
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
OR HIGH-PRESSURE LIQUID CHROMATOGRAPHY
• Is a chromatogrphic technique that can separate a mixture of compounds, and is used in biochemistry and analytical chemistry to identify , quantify and purify the individual components of the mixture.

• HPLC utilizes different types of stationary phase [ typically , hydrophobic saturated carbon chains ] .

• A pump that movse the mobile phase(s) and analyte through the column.
• A detector that provides a characteristic retention time for the analyte , the detector may also provide other characteristic information ( i.e. UV / VIS spectroscopic data for analyte if so equipped).
• Analyte retention time varies depending on the strength of its interactions with the stationary phase the ratio /composition of solvent (s) used and the flow rate of the mobile phase.

• With HPLC , a pump (rather than gravity).

• Provides the higher pressure required to propel the mobile phase and analyte through the densely packed column . the increased density arises from smaller particle sizes.

This allows for a better separation on columns of shorter length when compared to ordinary column chromatography.
Thank you
1