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Higher percentage of organic solvents gave an incomplete separation with some peaks co-eluting, while higher percentage of water in the mobile phase resulted in peak broadening and prolongation of the analysis. With an injection volume of 30μL, the peaks in the chromatogram were poorly separated, given that a high concentration of analytes can result in broad peaks and co-elution, while 10 μL were in the other hand too low. After that, we did experiments with several different mobile phases composed of either methanol and water or aceto- nitrile and water, where the percentage of the non-polar solvent ranged from 30 to 80%. |
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