I am interested in Thermo Scientific EASY-Spray 150umx15 cm LC Columns. I want to use the capillary column with the nanoLC system.
Using the capillary column with the nanoLC system, is it possible to achieve repeatability as in the Capillary-LC system? This is because, we have the Thermo Ultimate 3000 RSLCnano system with Proflow cell. The system allows flow up to 1.5 ul/min. So, is it possible to achieve maximum efficiency and high reproducibility in the nanoLC system by using a 20 µm nanoviper as in the capillary-LC system?
There is not any obstacle to using capillary flow columns with a nanoLC system in terms of configurational compatibility. However, using higher flows near the upper limit of the nanoLC system to deliver the total flow needed by the capillary column is not a suggestible option. This may cause frequent pump failure due to the usage of continuously at full capacity but more possibility is to see broader peaks. Even using the highest nanoflows, the capillary columns possibly demonstrate high peak broadening since the delivered flow by the nanosystem would be very low for its range of activity. Broadening means loss of chromatographic resolution which is nonsense for any proteomic and metabolomic analysis. Reproducibility would be OK but dramatically extended run times may be another drawback of the purposed platform.
Anyway, I would recommend adding a single makeup pump if you have one, in front of the proflow system by using a tee connection to increase total flow without disturbing the instrumental configuration and gradient profile...
Thank you for your answer. I am working on quantitative analysis. Therefore, the capillary column didn't extent my run time, on the contrary, with a good LC-MS method, I can see the relevant peptides in a short run time. I didn't have a wide peak problem, either. My only problem is that the RSD between runes is more than 10 percent. For quantitative analysis, using capillary column should have been more robust than nano column. However, I could not reach the robustness I imagined. I couldn't find any details that could cause the issue. I wondered if I was having this problem because I was using a capillary column in nano configuration. Thanks again for your recommendation. I will try it.
You may use the spike-in peptide approach by using labeled synthetic peptides to compensate and normalize the bias for reliable quantitative analysis if you cannot find any alternative.
Good luck
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