I am using Phenomenax rezex column with 5 mN H2SO4 as eluent flowing through the column at flow rate of 0.6 ml/min. RI temperature is 36C and column temperature is 60C. At the time of glucose standard injection, I am getting this type of response ( enclosed hereby). Eluents and standards has been properly prepared and degassed. What is the reason behind such type of response, kindly let me know.
Hi Aadarsh Madhu
Generally, Solution: Adjust or change the sample solvent. Dilute sample in mobile phase whenever possible. d) Mobile phase more absorptive than sample components to UV wavelength (vacancy peaks). Solution: Change UV wavelength or use a mobile phase that does not adsorb the chosen wavelength.
for more U can follow these 2 discussion
https://www.researchgate.net/post/The_appearance_of_Negative_peaks_after_positive_peaks_exiting_working_with_HPLC_RI_detector_Why_does_it_happen_and_how_can_I_remove_them
https://www.researchgate.net/post/Why_I_am_getting_negative_peaks_on_HPLC
and this link too
https://www.knauer.net/en/how-to-deal-with-negative-peaks/f19754#:~:text=Solution%3A%20Adjust%20or%20change%20sample,does%20not%20adsorb%20chosen%20wavelength.
Best Luck
Debanjan Chatterjee Thanks for your response! But we are not using UV detector We are using RI detector.
Even if the signal went to negative values, did you get a peak or not?
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