3. LC×LC method development
Whether offline or online, it is necessary to find a sufficiently orthogonal LC×LC combination. In offline LC×LC, the two dimensions can then be optimized independently of each other, the method development being that conventionally used in LC. In online LC×LC, development is more complicated, as the two dimensions are linked and must therefore be optimized simultaneously.
The first step in LC×LC method development is to select two chromatographic systems (figure
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LC×LC method development