Does the column overload amount differ for different types of columns? I am currently experiencing an issue of a rise in baseline until the peak elutes off, and then the basline goes back to normal. Any ideas why?
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By Anonymous on Tuesday, March 23, 2004 - 10:28 am:
I do not understand your baseline description but I can answer your first question. Yes, different stationary phases have different mass loading capacities. It's very difficult to say that brand X has higher loading than brand Y. Surface area is one criteria, but is not the only one. Plus you can load more analyte if the analyte is not charged.
What is your analyte, what is your pH and what is your column?
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By Anonymous on Wednesday, March 24, 2004 - 04:42 am:
Why does the basline rise after injection of sample and then go back to normal? .. (I hope I understand your question as you want to say.) .. For my idea, it is because, in sample you are going to injcet, there are some impurities or other substances or solvent in sample itself that differ from mobile phase (may be same type of solvent but different in concentration). These substances or solvent are not absorbed with stationary phase so they come along with mobile phase and diffuse in mobile phase. And also, These substances do not highly absorb UV light or highly response to detector you use, so they do not give as peaks but only rising baseline. After all injected sample is eluated the baseline back to normal. ... Hoping you understand what I want to describe.
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By Chris Pohl on Wednesday, March 24, 2004 - 02:13 pm:
In chromatography, overloading can result in a "fronting" peak such as the one you describe above, broadening in a symmetrical fashion or a "tailing" peak where there is a sharp rise at the onset of the peak with a gradual return to baseline. Each of these phenomenon is intimately connected with the relative affinity of your analyte and the eluent component responsible for the elution. The fronting phenomenon you describe is symptomatic of an analyte which has lower affinity for the stationary phase than your eluent species. Fronting occurs because at high enough loading levels the elution speed of the analyte slows down because the high concentration of analyte in the sample zone is sufficient to permit the analyte to play a role in the elution strength of the eluent in that zone. Since the analyte is a weaker eluent than your mobile phase elution species in this example, this causes the sample band to slow down as the concentration rises. After the concentration reaches the maximum, the process reverses, resulting in a nearly vertical trailing edge. Symmetrical increases in peak width occur when the eluent species and the analyte have nearly equal partition coefficients. Tailing peaks occur when the analyte species has higher affinity than the eluent for the stationary phase.