The retention time of analytes in HPLC is managed by the polarity of the mobile phase. The mobile phase of HPLC typically uses of nonpolar and polar mixtures/combinations of miscible organic solvents and water/buffers. Selection of proper mobile phase is an important aspect in HPLC method development. Before uses mobile phase and buffer it should be filtered throughout 0.5-micron and degas all aqueous and organic solvents to remove all particulate impurities and dissolve gases.
Check the compatibility, relative polarity, and miscibility of the solvents being used. Make use of only high purity solvents for mobile phase, and water should be HPLC grade or filtered and deionized, Generally transparent glass bottles use to storage of mobile phase because of glass is chemically inert and for visual inspection.
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