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Quantitative Chemical Analysis

Seventh Edition

Chapter 25 High-Performance Liquid

Chromatography

Copyright © 2007 by W. H. Freeman and Company

Daniel C. Harris

Suggested problems: Chapter 25: 1a, 15 Chapter 26: 25, 28

EXAM FOUR – TUESDAY APRIL 20th Separation modes and HPLC

Pumps and Injection valves for HPLC Sample introduction and HPLC

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Silica surface and bonded phases Particle size and separation efficiency

Selecting the right solvent for HPLC Isocratic separation of a mixture of aromatic compounds

“A” is 25 mM KH2PO4 and “B” is acetonitrile. Column is a 25 cm x 0.46 cm ODS with 5 µm silica.

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Isocratic separation of a mixture of aromatic compounds cont’d Isocratic separation of a mixture of aromatic compounds cont’d

Gradient elution of the same mixture! Detectors for HPLC

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Quantitative Chemical Analysis

Seventh Edition

Chapter 26 Chromatographic Methods and

Capillary Electrophoresis

Copyright © 2007 by W. H. Freeman and Company

Daniel C. Harris

Ion exchange resins and ion chromatography

Gradient elution and ion exchange chromatography

Higher the atomic # of the lanthanide, the smaller its ionic radius and the more strongly it binds to the resin

Ion exchange chromatography - analysis of pond water

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Indirect detection

Mobile phase contains 1 mM sodium phthalate

Ion-pair chromatography

Addition of sodium octanesulfonate to the mobile phase. Negative sulfonate groups act as anion-exchange sites for analytes such as protonated organic bases (BH+)

Schematic of an instrument for capillary electrophoresis Laminar flow versus EOF profile

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Separation efficiency in HPLC versus CE Indirect detection of isotopes separated by CE

Detection challenges in CE Electrochemical Detection in CE

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Separation of neutral compounds by MEKC Integrated biochip for DNA analysis