Dear colleagues,
I am analyzing reducing sugar content of several breeding lines of potato in HPLC. The mobile phase I am using is 80:20 acetonitrile:water and I sonicate it for 5 min to get rid of air bubbles. The column name is Inertsil NH2 and its conditions are 40oC, 1ml/min. I have tried several extraction methods (with only water, asetonitrile:water, methanol and ethanol) and some are already present in articles (the same column is used). I have decided to continue with ethanol and here I attached the result of varying ethanol water concentrations. The first chromatogram shows the results for only one genotype and the second chromatogram shows two genotypes which are 50. and 40. genotypes. Actually, after several trials, I have succeed to get proper peaks however at this time I have confronted baseline noise. I appreciate your helps on this topics.
Baseline noise has many causes and solutions. Often it is due to no degassing or poor quality degassing (use Helium sparging or an inline HPLC degasser for best results. Sonication for five minutes is VERY POOR at degassing mobile phase solution. The gas just bleeds back into the solution over time (~20-30 minutes) and results in drift.
I have attached an article link on the topic of baseline noise which may help you understand, troubleshoot and solve this problem; "Common Causes of Baseline Noise in HPLC, UHPLC".
Good luck with your project!
http://hplctips.blogspot.com/2014/09/common-causes-of-baseline-noise.html
Some of the small baseline disturbances you might be referring to as noise are seen at the same RT in all of your chromatograms, so they are probably real components in the samples. If you are concerned about the drift, RI detectors can be very sensitive to variations in solvent and temperature. As already mentioned, degassing properly is very important. Also, make sure the room temperature is reasonably well controlled and you allow enough time for the detector internal temperature and column oven to equilibrate.
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