By Hansen, Steen; Pedersen-Bjergaard, Stig

ISBN-10: 1118716833

ISBN-13: 9781118716830

ISBN-10: 1118716868

ISBN-13: 9781118716861

ISBN-10: 1301351431

ISBN-13: 9781301351435

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Extra info for Bioanalysis of pharmaceuticals : sample preparation, separation techniques and mass spectrometry

Sample text

When the pH is below the pI, the net charge is positive. The value of the pI can be calculated from the pKa values from all the side chains with acid–base properties as well as the pKa values of the N- and C-terminal groups. The calculation itself will not be discussed in this textbook. However, there are several web-based pI calculators available. EPO contains many functional groups with acid–base properties: 25 basic ones (3 histidine, 13 arginine, 8 lysine, and 1 N-terminal amine) and 20 acidic ones (6 aspartic acid, 13 glutamic acid, and 1 C-terminal carboxyl).

Similarly, a decrease in internal diameter of a GC capillary column will increase the efficiency. 1 Chromatographic Principles Normal Phase Chromatography Normal phase chromatography, also called straight phase chromatography, is the most common separation principle in TLC but can also be performed in HPLC mode. A polar stationary phase is used along with a more nonpolar mobile phase. A typical choice could be the use of bare silica as the stationary phase and a heptane–ethyl acetate or heptane–propanol mixture as the mobile phase.

Analytes with no affinity to (no interaction with) the polar stationary silica will not be retained and will travel with the speed of the mobile phase. Analytes having polar functional groups will have a higher affinity to the polar silica and will show retention in the system. The adsorption onto the stationary silica is a reversible interaction, and an increase in the polar component of the mobile phase will increase the competition between analyte molecules and polar molecules from the mobile phase for the adsorption sites on the surface, thus weakening the interaction of the analyte with the silica.

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Bioanalysis of pharmaceuticals : sample preparation, separation techniques and mass spectrometry by Hansen, Steen; Pedersen-Bjergaard, Stig

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