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A novel LC method was applied to enhance the peak resolution of two ecdysteroids: 20-hydroxyecdysone and its 3-epimer. An isocratic solvent system of methanol–water (6:4, v/v) was used on a C18 column, and 100 μL of water was injected during the development in such a manner that the eluted solvent peak appeared exactly between the two overlapping peaks. This resulted in the increased retardation of...
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