Ultra Performance Liquid Chromatography

[chrommgr]

Greetings. This is my first posting to this forum. To all those who pratice HPLC on a daily basis, and for those of you who are lab managers of a chromatography lab, I was wondering if I could get your professional opinions of a relatively new development in liquid chromatography, something that Waters Corporation has dubbed "UPLC".

Perhaps you have heard about it. If you have, or better yet, if you currently use UPLC in your lab, would you consider these questions:

1. Does your lab have plans to purchase a UPLC (Waters is the only current vendor) in the near future?

2. What are the motivating factors behind a UPLC purchase if you currently already have HPLC? Is it simply the "need for speed" or is there anything else at play here?

3. How would you describe the market need for UPLC? Weak, moderate or strong? Please explain your answer.

4. How likely would your organization utilize a 3rd party testing lab for your analysis needs if that 3rd party lab had UPLC available, but your lab did not yet have it available.

These will probably suffice for now. I may have some follow-up questions later on. I appreciate your responses.

chrommgr

[Apharmd Battler]

A couple of our development labs did some evaluations last year, and I know in my old lab we had one on loan from Waters to play around with. I'll try to sift through my desk and e-mail and report the findings done by at least two of our labs. I remember the findings were favorable.

[aldij]

please, could you also send me a copy of the report? we are in the process of selecting a new lc-ms-system and uplc looks like an interesting option.

aldij

[Apharmd Battler]

I can't to seem to find the documents at the moment. Also, their is some concerns with proprietary info and such. Got a bunch of irons in the fire now, but I'll dig around my network folders later today in between runs and try to come up with something. I'll stay in touch.

I know I've got some stuff lying around, but I know there was a big spread just a few months ago in LC/GC, you can probably find this online. Several scientists did some work and posted published their result in the attached insert.

<Edit>
I found one of the reports and here were the conclusions:

Conclusions
•OQ gradient data comparable to Agilent 1100
•Injector precision (partial loop) and baseline
noise issues a major problem
•Advantage of injector wash system inconclusive
•HPLC mode of operation on Acquity was
comparable for 4 gradient impurity assays, and
not comparable for 2 isocratic assays.
• Retention time mismatches potentially due to
Flow/temp. issues
•In general, UPLC mode provides up to six times
improvement in run time with minimal tradeoff in
resolution
•UPLC mode showed sensitivity improvements,
allowing lower volume injections necessary to
prevent overload of 1.7 ?m columns.
•UPLC offers 2 times improvement in run time
over Agilent 1.8 ?m technology with
improvement in resolution.
• Dependent on assay needs utilizing an entire instrument
to obtain this improvement may not be justified.

I hope this helps, I would post more but then some of the stuff evaluated is from our early phase pipeline, and I don't know if it's safe to post publicly. I keep digging. I hope this info at least helps you in some way.

[Mattias]

The UPLC technique looks very interesting, and I appreciate that Waters really has made an effort to develop an optimised system.

I work with method development in a development lab, and our methods are transferred to QC labs around the world. It is not likely that all these labs will replace their LCs with Aquiety systems (which also cost about 50% more than a Dionex system, for example).

I think the big market (which is the pharmaceutical buisness) will be limited for a long period of time.


But I would really like to try one

[gemm]

we have a waters uplc for integration in our workflow right now. we plan to use it for ht-screening, so speed is essential. to answer your questions:

1. yes, but waters is not the only vendor.

2. it is simply the "need for speed". this means the cycle time from injection to injection and not the time for the chromatografic run only.

3. moderate, but growing. for LCMS application, the requirements for the MS are demanding to keep up with the UPLC. TOF or similar technique is required for reliable detection of the narrow peaks (full scan). existing LC and MS equipment has to be exchanged/upgraded. this is usually not a one-day-decision.

4. not at all. evaluation and analysis is done in house. this includes integration of hardware+software in the existing workflow and usage real life samples instead of test mixtures. visiting other labs to share experience might be an option.

finally some aspects you might consider about the waters uplc: the injector is not (yet) capable of drawing a second sample, while the first one is still on the column. you have to wait until the chromatographic run is finished. this results in a longer cycle time.
the injector is not capable of dissolving samples by drawing solvent from a reservoir. so you have to use sealed plates or taking the risc of solvent evaporation from your plate (we use 384 wellplates).

in general, high speed (or high pressure) LC is done at higher temperatures. this (and the pressure) reduces the lifetime of your columns. some vendors try to compensate this by launching new packing materials, but i did not get the chance to evaluate these yet.


@Apharmd Battler

UPLC offers 2 times improvement in run time over Agilent 1.8 ?m technology with improvement in resolution.

i cannot agree with you in this. did you compare cycle time or just runtime ?

[Apharmd Battler]

Gemm, this was work performed at an affiliated site, let me get back to you on that after I do some digging.

[james little]

I have a Acquity on my Quattro Micro. Primarily doing LCMS on drugs in plasma.

I am using a using a 3.5 u (2.0 x 50) reverse phase column. It gives me better peak shapes than the Acquity columns I have tried to date.

I Like the system. Retention times with fast gradients really very reproducible.

I have had better results (better precision) with 2 ml vials with preslit tops compared to 96-well with silicone/Teflon coated on one side). Waters suggests using other types of tops now, but I haven't tried yet.

I am doing MRM on only a few analytes at a time, so able to get adequate number of samples across peak.

My runs are usually 4.5 minutes total including the autosampler sequence of about 30 seconds. This is a gradient from 80% aqueous to 100% organic.

I just include my sampling time of 30 seconds in the equilibration time, so not really a problem. COuld be a limitation if doing isocratic.

Some examples of chromatograms shown in the top topic on my web page on matrix effects in LCMS analyses.

See..

http://users.chartertn.net/slittle/

[ravi]

..............•UPLC offers 2 times improvement in run time over Agilent 1.8 ?m technology with improvement in resolution..................

It funny, but I'm having the hardest time getting a particle distribution number from Agilent for their 1.8µ columns. The same with Thermo. While Waters provides it on the spec sheet.

I'm wondering if their 1.8µ claim is a little ambitious.