By Johan Lindholm
This thesis issues the improvement and validation of excessive functionality liquid chromatography (HPLC) equipment aimed for 2 industrially vital parts: (i) research of biotechnological synthesis and (ii) decision of adsorption isotherm parameters.
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Extra info for Development and Validation of HPLC Methods for Analytical and Preparative Purposes
These sites, type-II sites, are responsible for the enantiomeric separations. On most CSPs the type-II sites are relatively few. 3 The Langmuir Isotherm The isotherm that is most easily understood theoretically and widely applicable to experimental data is, known as the Langmuir isotherm. This is also the simplest model of a nonlinear isotherm. It accounts well for the adsorption of single components on homogenous surfaces at low to moderate concentrations, or for the adsorption to isolated type-I or type-II sites [103, 110, 111].
At the first arrow a small sample containing the component is injected into the column that is only equilibrated with pure mobile phase lacking the component. In the following steps the column is equilibrated at different concentrations of the component and small amounts of pure mobile phase lacking the component are injected at the times marked with the arrows. At higher plateau concentrations the retention time is decreased caused by the non-linearity of the adsorption isotherm. 56 Figure 10.
At least four of the six QC samples should be within 20% of there respectively nominal value, and at least one at each concentration level [16, 89]. Also a standard curve should be processed during each run . 9 Validation of Biotechnological Synthesis. V. values < 15%. V. values < 2% . This is possible, since in the latter case the matrix is typically much simpler and the analyte concentration can be chosen freely, so that extremely low/high concentrations can be avoided. There are no detailed recommendations for analytical procedures in the field of biotechnological production of drugs, in contrast to the recommendations made by the FDA for bioanalytical methods .
Development and Validation of HPLC Methods for Analytical and Preparative Purposes by Johan Lindholm