Authors
Benjamin Ingham
The University of Manchester
Objective
Apply functional data analysis and functional design of experiments (FDOE) for the optimization of an analytical method to allow for the accurate quantification of two biological components.
Background
Bob is attempting to produce a high-performance liquid chromatography (HPLC) method that can separate two closely related sophorolipid biosurfactants (C18:1 and C17:1) that co-elute (appear close together) on the chromatogram – making quantification difficult.
To have a suitable HPLC method, Bob must be able to:
- -Sufficiently separate the two peaks.
- -Ensure that the peaks are sharp (small in width) and tall (high in sensitivity).
The Task
Bob is entrusted with the following tasks:
- -Apply statistical design of experiments (DOE) to find HPLC settings that can improve the method.
- -Use functional data analysis to understand how the curve shape changes as factors change.
- -Identify optimized conditions for the separation of the two peaks.
To improve the separation of the two peaks with the HPLC, two control parameters were chosen to be changed: the mobile phase flow rate and the column temperature. The temperature was altered to 30, 35, or 40°C, and the flow rate to 0.7 or 1.4 mL/min.