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Chromatographic conditions, especially the composition and nature of the mobile phase, usage of different columns, and different extraction methods such as solid phase, precipitation, and liquid-liquid extraction methods were optimized through several trials to achieve the best resolution and increase the signal of SF and SFD5.
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Two mobile phases were used: mobile phase (A) 0.05% (v/v) TFA and mobile phase (B) 100% acetonitrile.
Modifications in mobile phase concentration, mobile phase volume, mobile phase saturation time, and temperature were examined to check the robustness.
The mobile phase was composed of pet.
flow rate of mobile phase (mL/min).
The mobile phase was acetonitrile (65%).
Water was used as the mobile phase.
NH4OAc and MeOH as mobile phase.
Moreover, mobile phase stability was also checked.
The mobile phase used was pure acetonitrile.
ethylene glycol/acetone (5 3 1) as mobile phase.
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