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Hello
someone can help me ? I work on HPLC and I have the base line less than zero knowing that I work on the purification of DNA primers using a Hamilton PRP1 column and a mobile phase trietyl ammonium acetate and acetonitrile
Hello
someone can help me ? I work on HPLC and I have the base line less than zero knowing that I work on the purification of DNA primers using a Hamilton PRP1 column and a mobile phase trietyl ammonium acetate and acetonitrile
please help me
thank's
Please send Chromatograph may be some linkage in pump column parts please run blank sample one time
I'm assuming UV detection? A negative baseline suggests the system set the baseline to zero while there was an adsorbing material in the flow cell. This material could be a particle or precipitate, possibly an air bubble, or more likely, a late eluting peak or some solvent which absorbs UV. As this gets washed from the flow cell, the absorbance is less than that measured is less than the absorbance measured to set the baseline, so the baseline is negative. To avoid this, add a wash step at the end of your run to wash off non-polar impurities, and you may require a longer equilibration so the starting baseline may be correctly measured.
A chromatogram would be helpful as well. Does the chromatography of your primers seem fine?
Hello, thank you for your answer
here is attached two image showing the on-line signal and offline
the peaks during the injection are always negative even the signal is always under the zero
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