Dear colleagues,
With access to only one Orbitrap instrument, we are conceiving 2 different LC systems for switching between proteomics (nano-UHPLC) and metabolomics (UHPLC) analyses once a month.
Changing of UPLC columns are thus expected to be routine. From various vendors, there are 2 main types of UPLC connections / fittings:
A vendor has a new type of UHPLC system, using (1) PEEK sleeve columns. In the vendor's tutorial online, however, the PEEK sleeve columns (re-connection and set-ups) seem a bit cumbersome (e.g. as it contains quite many small parts, involving various tools).
On the other hand, the (2) nanoViper offers a pre-assembled "tool-free", "finger-tight" fitting, which is more straight forward (without the meticulous extra care not to over tighten and damage the fused silica columns).
a). Would you mind sharing your experience regarding the usage of (1) PEEK sleeve, and/ or (2) nanoViper column fittings? How easy / difficult for the respective type of connections to achieve "dead-volume free" ?
b). We would also appreciate it a lot if you could advice regarding the consideration of UHPLC (especially nano-UHPLC) systems [e.g. between vendor(s) that uses the (1) PEEK sleeve, versus another using (2) nanoViper column fittings]?
Thank you very much!
I have to admit, I'm a big fan of the nanoViper connections, they a really are easy to use. Tool free, high pressure connections, nice. That said, it really isn't that difficult to deal with PEEK. I'm not sure I fully understand what you mean by "PEEK sleeve". If you mean using PEEK sleeves around capillary silcia tubing, then we are on the same page. If you don't mean that, then I guess we aren't. I'll assume you do mean the PEEK around silica. With some experience, these are not difficult to deal with, most of our UHPLC systems use this type of connection. If your users are likely to be less experienced, I'd think they'd adapt to the nanoVipers more readily.
I like the nanoViper fittings. I use them for both high (metabolomics) and low-nano (proteomics) flow UHPLC applications. They are easy to use, require no tools and we have no leaks. I use them with Dionex, EasyNano and Agilent 1290 LC systems with the Orbitrap Fusion and even a 6490 QQQ now.
Dear Gabriel,
Thank you very much for your advice!
Would you mind sharing the type of UHPLC / nano-UHPLC systems you coupled with the Orbitrap Fusion for proteomics and metabolomics analyses?
Are you also switching between proteomics (nano-UHPLC) and metabolomics (UHPLC) analyses on the same Orbitrap Fusion instrument? How frequent would such a switch be (e.g. once a month or longer)?
Appreciated !
Viper are good with high flow systems in my opinion, they are nice with nano-uhplc's as well for proteomics, the only downside is you can't pack your own columns and require ez-spray and Thermo/Dionex prepacked columns which are expensive. PEEK or just silica tubing is fine if you're using the nano-uplc1000 systems.
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