I am wondering if anyone that is familiar with chromatography could kindly give a brief introduction of this technology and describe the differences between regular HPLC and this technology? Also, which brand of nano-HPLC/UPLC is holding a good reputation?
The term "nanoLC" refers to flow rates in the nanoliter/min range (as opposed to high microliter/min to low milliliter/min range for regular HPLC). It is usually used for separating minute sample amounts with concentration-sensitive detectors, mostly nanoSpray mass spectrometry. A typical nanoLC/MS/MS setup would be a 75 micrometer I.D. column at a flow rate of 200-350 nanoliter/min.
High quality nanoLC (or, if you use long or small particle columns, nanoUPLC) vendors include Waters, Dionex, Eksigent and Proxeon. All of these can work well in nanoLC/MS/MS setups. If you are looking into purchasing a system, I would strongly recommend to look into the vendors' software integration with your MS software, as well as the availability and reputation of a regional service and support team.
Nano HPLC/UPLC is a relatively new development in chromatography world driven by recent advancements in proteomics that would require decreasing of inner diameter (ID) of liquid chromatography (LC) column to allow for a smaller sample amount and to increase sensitivity. In nano HPLC or UPLC is better than usual HPLC because minimal samples loss, higher sensitivity, separation efficiency and peak capacity is high, perfect of handling minute or dilute samples, superior coupling to MS, flow rate is nL/min to microlitre/min and overall it is used capillary column instead of normal HPLC column.
UPLC is usually reserved for systems that go to very high pressures, around 10,000 psi. HPLC, on the other hand, tends to max out around the 4,000-6,000 psi mark. The UPLC systems are capable of using chromatography resins with a diameter < 2um. It is less about the ID of the capillaries used and more about the particle diameter and the pressures involved. These then lead to higher peak capacity and superior separation. There is really no difference in flow rates between nanoflow-UPLC and nanoflow-HPLC systems.
Hope that helps,
Peter
The term "nanoLC" refers to flow rates in the nanoliter/min range (as opposed to high microliter/min to low milliliter/min range for regular HPLC). It is usually used for separating minute sample amounts with concentration-sensitive detectors, mostly nanoSpray mass spectrometry. A typical nanoLC/MS/MS setup would be a 75 micrometer I.D. column at a flow rate of 200-350 nanoliter/min.
High quality nanoLC (or, if you use long or small particle columns, nanoUPLC) vendors include Waters, Dionex, Eksigent and Proxeon. All of these can work well in nanoLC/MS/MS setups. If you are looking into purchasing a system, I would strongly recommend to look into the vendors' software integration with your MS software, as well as the availability and reputation of a regional service and support team.
- Badges
- Science method