Chromatography Forum

Dear all,

I am planing to purchase a UPLC instrument and I am wondering about the life time of sub2 micro columns that used in UPLC, is it similar to ordinary HPLC columns?[/size][/size]

In regard to the column lifetimes:

Not every sub-2 is equal. Stick to the companies that have been manufacturing them for a while, like Agilent and Waters. I've used two columns from two other column manufacturers (besides Waters and Agilent) - one a true sub-2, the other a 3-micron running at high pressure (ostensibly within the acceptable pressure range for the column), and had them die within about 100-125 injections during method development. Meanwhile, I have an Agilent ZORBAX RRHD Extend C18 3.0x100mm 1.8-micron that I've been running for months at 650-750 bar with real environmental samples and no guard column, just a 0.2-micron in-line Phenomenex KrudKatcher filter, and I've had no problems in terms of column lifetime. There are probably 1000-1500 injections on this column by now, which to me is well within the range of acceptable lifetime for any column, especially without a guard column.

A caveat about new instruments in general, and this is my experience only, so I can't say that you'll encounter the same problems, or even need to consider the same issues, but I want to say my piece anyway...

1. Make sure there are an adequate number of service personnel in your area before you buy whatever you end up buying.

2. Get a service contract. It might take you a while to learn the new instrument, and the difference between breaking it due to your own ignorance while within the coverage of a service contract and outside the service contract can be substantial. Also, some manufacturers are so new to UHPLC/Fast LC/whatever the new euphemism is, that their hardware is not fully tested in the ways that a real chromatographer will test it (read: break it). Trust me - I have an Agilent engineer coming out Monday to replace the pump drives on one of my Agilent 1290 binary pumps (not my fault, I promise - but I'm glad it's under contract).

3. Make sure your new system is compatible with your old data quantitation/acquisition software. If not, make sure that any new software you might need fits into your current workflow scheme - you don't want to get a fancy new instrument, only to realize the data it generates is incompatible with either your LIMS system or your quantitation (quantification? ) program.

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thanx a lot "bisnettrj2" for the useful information and tips. as you said; we are planning to get a service contract and also a training courses on the instrument and the software.
thanx again

Hello. Also depends on the pressure - if you do not push with terrible force, and use pressure to 300 bar, the column can be a very long time. But it will have to pay for the time of analysis, which is not always acceptable.

New to this forum, first post here. [/intro]

I attended a Waters UPLC seminar a few months ago after our lab purchased their new UPLC instrument. It was mentioned that the temperature of the column has a large effect on its lifetime. Operating UPLC columns at temperatures above room temp (25C) severely decreases their lifetime. Otherwise, column lifetime for UPLC is pretty comparable to HPLC as long as you stay away from the upper bound of their pressure limits and harsh solvents.

shornets45:

The Acquity BEH130/BEH300 manual recommends 20-55 C, and the Acquity BEH manual recommends 20-90 C operating temperatures. Nowhere in the manual does it state that operating above 25 C is detrimental to column lifetime; the only caveat is that running at extremes of temperature, pH, and pressure will have an impact on column lifetime (which is the same for any column). In fact, the cleaning guideline for the BEH130 columns recommends 35-55 C to increase column cleaning efficiency, which would seem to be in direct opposition to your statement about temperature - why would Waters recommend a temperature detrimental to column lifetime in their care and use manual? Do you have a publication or any documentation of the "25 C limit" to back up your statement?


http://www.waters.com/webassets/cms/sup ... 001465.pdf
http://www.waters.com/webassets/cms/sup ... 001371.pdf

Just a word about sub-2um column life time: I have been using Waters sub-2um columns for 6 years now and I find it satisfactory. My current column has had 2500 injections and is still alive. The previous one died after 1300 injections. Both were used at pressures from 350-800 bars and temps from 25-60 °C. I usually inject dirty stuff (plant extracts). I admit I often perform cleaning procedures.

You *do* have to be more careful about "chromatographic hygeine": filter all mobile phases and samples, watch the expiration dates on aqueous buffers, be extra careful with fittings, etc.

shornets45:

The Acquity BEH130/BEH300 manual recommends 20-55 C, and the Acquity BEH manual recommends 20-90 C operating temperatures. Nowhere in the manual does it state that operating above 25 C is detrimental to column lifetime; the only caveat is that running at extremes of temperature, pH, and pressure will have an impact on column lifetime (which is the same for any column). In fact, the cleaning guideline for the BEH130 columns recommends 35-55 C to increase column cleaning efficiency, which would seem to be in direct opposition to your statement about temperature - why would Waters recommend a temperature detrimental to column lifetime in their care and use manual? Do you have a publication or any documentation of the "25 C limit" to back up your statement?


http://www.waters.com/webassets/cms/sup ... 001465.pdf
http://www.waters.com/webassets/cms/sup ... 001371.pdf

Unfortunately I do not. These were Waters employees giving a seminar and the question was brought up. I will see if I can get some clarification from my contact there.

Are you sure that the manuals weren't defining "safe-operating zones"? I'm talking about column lifetime, not temperature limits. Certainly I agree that a higher temperature would increase the cleaning efficiency, however prolonged use of the column at elevated temperatures (40-50C) decreases the overall lifetime.

Again, this was the Waters response to this question at one of their system care seminars.

They are defining safe operating zones, I believe; however, I'm not sure anyone would describe 35-45 C as an "extreme temperature" terms of an HPLC column, as long as you're not talking about an extremely low or high pH buffer in combination with the temperature. If you combine high temps (see Table 3 in the BEH manual) AND aggressive pH values, then you will get shorter column lifetimes. I've never personally seen any issue with using my UPLC columns at 40-50 C, but I don't run buffers on these columns (just water:methanol), so maybe the case would be different if I was. In any case, I think operating at 'safe' ranges is a great idea, but I think 'anything over 25 C' is an unrealistic definition of 'safe'. That's like saying I get the best lifetime out of my columns if I never use them!