Discussion
Why must the mobile phase be transparent at the detection wavelength?
Answer: D
A mobile phase absorbing at the working wavelength leaves too little light for the small peaks that matter, and gradient elution would produce a drifting baseline as its composition changes. Reactivity, retention and cell damage are separate concerns. That the choice of solvent is constrained by the detector as well as by the separation is a coupling between two parts of a method that catches people out when a wavelength is changed without revisiting the eluent. Working one numerical case through both cases side by side fixes the distinction better than any rule of thumb.
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