Chromatography Forum

Hello guys,

I need your comments or advice on which Fast LC is the best in the market, especially if it is to be attached to an API (Appliebiosystems) mass spetrometer. The three systems that we are considering are the Waters UPLC, Agilent 1200 (rapid resolution) and the ThermoFisher fast LC system. If there's anyone among you who have any of these LC systems hooked up to an API mass spectrometer, I would really appreciate all your comments and advice, base on your experience with these systems. Thanks!

In our lab we are using either RRLC from Agilent and RFLC / Prominence
from Shimadzu and we are very happy about the fast seperations.

We have not got very good feedback from either water's UPLC and Thermo's
as well

I can confidently suggest you to go either with Agilent or Shimadzu

We have 9 Waters Acquity UPLCs (all used in conjuction with various Waters/Micromass mass specs). We love them. We are even thinking about putting them on the front of our ABI QTraps and 3200 (we have a shimadzu on the front of the 3200 and it is a piece o' junk).

In my opinion, they are the only way to go if you want 'true' UPLC, not the UHPLC or UFLC stuff. Hit me back if you want some more info. on what we do with them.

Dear Sindhu and Stryder08, thank you for your replies.

Sindhu: I would like to ask you about your Agilent SL (or RRLC?). How long have you used it? Are you using it mainly for Very High Pressure LC (VHPLC)? Have you tried using it for HPLC operations? Did you have any problems transferring methods from HPLC to VHPLC? Is it true that there is a need to do physical reconfiguration of the pump when switching between HPLC and VHPLC? Is it user friendly in terms of troubleshooting and repair?


Stryder08: With the Waters UPLC, do you need to reconfigure physically the pump when switching from HPLC to VHPLC and vice versa? Any (minor or major) problem that you have encountered? frequency? Is it user friendly in terms of troubleshooting and repair? You mentioned about planning to attach one of your UPLC at the front end of an ABI mass spectrometer. Is it really doable and do you know of any users who has already done?


My apoligies for so many questions. We are planning to buy a VHPLC and need to be careful because the cost is high.

Thank you so, so, very much!

The big problem for Agilent HPLC is the LARGE dead volume. If you run fast gradient, you will see troubles.

Dear Sindhu and Stryder08, thank you for your replies.

Sindhu: I would like to ask you about your Agilent SL (or RRLC?). How long have you used it? Are you using it mainly for Very High Pressure LC (VHPLC)? Have you tried using it for HPLC operations? Did you have any problems transferring methods from HPLC to VHPLC? Is it true that there is a need to do physical reconfiguration of the pump when switching between HPLC and VHPLC? Is it user friendly in terms of troubleshooting and repair?


Stryder08: With the Waters UPLC, do you need to reconfigure physically the pump when switching from HPLC to VHPLC and vice versa? Any (minor or major) problem that you have encountered? frequency? Is it user friendly in terms of troubleshooting and repair? You mentioned about planning to attach one of your UPLC at the front end of an ABI mass spectrometer. Is it really doable and do you know of any users who has already done?


My apoligies for so many questions. We are planning to buy a VHPLC and need to be careful because the cost is high.

Thank you so, so, very much!

I don't know anything about switching between HPLC and UPLC, but all I would assume that you would have to change is the column to a larger particle size versus the smaller ID columns. I don't think you have to physically do anything.

When you have the ability of UPLC, why switch back to normal HPLC? In our industry, you don't. Unless you already have a validated method, I guess. We have revalidated most of our older HPLC/LC/MS/MS methods for UPLC or UPLC/MS/MS.

The Acquity UPLC is very easy to perform maintenance when needed. Even though not much maintenance is needed. Check valves rarely go bad.

The only downside that I have seen is the touchiness of the sample organizers (add-ons). They still have quite a few bugs.

Yes, putting an Acquity UPLC on the front of an ABI 3200 or QTrap or 4000 is very doable. I don't think ABI likes to admit it, but it is. We had a demo. for the UPLC on the front end of an ABI and really liked what we saw. In the not so distant future, we will be replacing the front ends with UPLCs.

Thank you stryder08 for your response. I am not more at peace knowing that UPLC can really work with the API2000.

Regarding your question as to why we need to resort back to standard HPLC runs when we have a UPLC, the reason why I am asking about switching back and forth from HPLC and VHPLC is because we are only a small lab and only have an API2000 MS at the moment. Due to budget constrainta, we intend to use the UPLC for now in standard LC runs with the API2000. We will only use the VHPLC capability of the UPLC once we acquire a faster scanning MS. So, there may be some period after the acquistion of the new faster scanning MS that we may need to switch back anf forth between HPLC and VHPLC.

Thank you stryder08 for your response. I am now more at peace knowing that UPLC can really work with the API2000.

Regarding your question as to why we need to resort back to standard HPLC runs when we have a UPLC, the reason why I am asking about switching back and forth from HPLC and VHPLC is because we are only a small lab and only have an API2000 MS at the moment. Due to budget constrainta, we intend to use the UPLC for now in standard LC runs with the API2000. We will only use the VHPLC capability of the UPLC once we acquire a faster scanning MS. So, there may be some period after the acquistion of the new faster scanning MS that we may need to switch back anf forth between HPLC and VHPLC.

...the reason why I am asking about switching back and forth from HPLC and VHPLC is because we are only a small lab and only have an API2000 MS at the moment. Due to budget constrainta, we intend to use the UPLC for now in standard LC runs with the API2000. We will only use the VHPLC capability of the UPLC once we acquire a faster scanning MS. So, there may be some period after the acquistion of the new faster scanning MS that we may need to switch back anf forth between HPLC and VHPLC.

This very much sounds like a system optimized for standard LC that can also handle UHPLC is the better choice for you. In addition, it will be the cheaper solution.

The Shimadzu Prominence UFLC will provide excellent fast LC and conventional LC. There is a higher pressure version ~9600 psi. The system has extremely low carry over. I have found the Shimadzu XR column at 2.2 micron to be very versatile for RP separations.


Stryder08- stating that a particular LC is not performing in the manner expected is not useful information for this forum. If you have some FACTS that suggest the Shimadzu does not perform to specification than let’s hear it.

The Shimadzu Prominence UFLC will provide excellent fast LC and conventional LC. There is a higher pressure version ~9600 psi. The system has extremely low carry over. I have found the Shimadzu XR column at 2.2 micron to be very versatile for RP separations.


Stryder08- stating that a particular LC is not performing in the manner expected is not useful information for this forum. If you have some FACTS that suggest the Shimadzu does not perform to specification than let’s hear it.

Well, I'm not here to argue, but the proof is in the pudding. We have 1 Shimadzu Prominence UFLC coupled to an ABI 3200. We have 6 Waters Acquity UPLCs couple with TQDs, 1 Waters Acquity UPLC coupled with a Quattro Premier XE, and 2 Waters Acquity UPLCs coupled with LCT Premier XEs. We have 2 Agilent 1200s coupled with ABI 3200 QTraps, but will be replaced with Acquity UPLCs in the future...what does that tell you? The lone Shimadzu isn't peforming up to snuff.

Hi Stryder08,

I am just curious. Is it that you find Waters UPLC better than Agilent 1200, that's why your lab plans to switch the two Agilent 1200 with the Waters UPLC as the front end for your ABI mass spectrometers?

Thanks!

Hi Stryder08,

I am just curious. Is it that you find Waters UPLC better than Agilent 1200, that's why your lab plans to switch the two Agilent 1200 with the Waters UPLC as the front end for your ABI mass spectrometers?

Thanks!

Yes, we find that the Waters Acquity UPLC is the best high pressure system out there. It is true UPLC, not UFLC, UHPLC, etc. Very little maintenance needs to be completed (only changed check valves one time in a little over 1.5 years, other than PMs, for 10 systems). The only downside that I have seen is the sample organizer; Waters really needs to redesign the thing.

Thank you all so much for all your helpful comments and suggestions. We have finally decided to purchase the Water UPLC system!

Hi Stryder08,

I am just curious. Is it that you find Waters UPLC better than Agilent 1200, that's why your lab plans to switch the two Agilent 1200 with the Waters UPLC as the front end for your ABI mass spectrometers?

Thanks!

Yes, we find that the Waters Acquity UPLC is the best high pressure system out there. It is true UPLC, not UFLC, UHPLC, etc. Very little maintenance needs to be completed (only changed check valves one time in a little over 1.5 years, other than PMs, for 10 systems). The only downside that I have seen is the sample organizer; Waters really needs to redesign the thing.

Was at a Waters seminar yesterday and got talking with the specialist. It seems that the sample organizer's cooling system isnt powerful enough to either maintain temp or reduce temp quickly once the door is opened then closed. Is this the probem you're experiencing?