EFFECT OF INTEGRATION PARAMETERS ON HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD DEVELOPMENT AND VALIDATION

An HPLC method is defined as a specific setting of the physical, chemical and data processing parameters which control the chromatographic analysis. During the development or validation of a method no special attention is usually given to the data processing parameters. However, it turns out that HPLC methods for complex samples can be very sensitive to minor changes in certain numerical integration parameters such as the threshold. A series of statistically designed chromatographic runs for a dyestuff with a very large number of peaks is presented where the interpretation of the data depends crucially on which threshold parameter value is chosen. It is therefore recommended that for development or validation of an HPLC method, especially in the context of quality control of complex substances, greater attention should be paid to the integration parameters.