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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 30¦ÌL, 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 ¦ÌL 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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