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Reverse phase High Performance Liquid Chromatography (RP-HPLC) is an analytical technique used to separate a variety of mixture based on their polarity. The “reverse phase” refers to non-polar stationary phase used in the HPLC, which traditionally has a polar stationary phase. Usually silica beads coated with non-polar alkyl chains act as a non-polar stationary phase. As sample is injected in the system and runs through the column, the non-polar components of mixture interact strongly with stationary phase and have higher retention times relative to polar components. The detector generates a chromatogram illustrating the concentration and retention time of each component of sample. (Credit: Muhammad Atif Khan)
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