Re: AAA, perspective?

Deb McMillen (mcmillen@morel.uoregon.edu)
Thu, 10 Jun 1999 10:54:23 -0700 (PDT)

James, Can the 507 autosampler be programmed for partial loop loads? And
does that give you the accuracy that you need for your validated method?
The partial loop loads on our 506 use just that amount of volume--full
loop loads use 1.5 x (the volume to be injected) + 50 microliters. Even
on a standalone loop you need to
overfill the loop to be assured that it is 100% filled with your sample,
due to adhesion (along the walls of the tubing) of the material that was
in the loop before you load your sample. It is just the nature of the
beast. Partial loop loads most likely are not truly injecting the sample
at the concentration that the sample is sitting in the autosampler vial.

One thing you can do, if your chromatography will tolerate it, is dilute
your sample and use a larger loop--the larger the loop in a way the less
sample you use--a 20ul loop uses 80 ul on this formula, 4 times the
sample; a 50 ul loop uses 125 ul; and a 100ul loop uses 200 ul, 2 times
the sample.

Hope this is helpful,
Deb McMillen
Institute of Molecular Biology
University of Oregon
Eugene OR 97403

On Wed, 9 Jun 1999, Freedy, James [OBI] wrote:

> Greetings,
>
> I am involved in amino acid analysis of a particular glyco-protein,
> for release testing. My efforts are in support of a validated method.
> Currently our QC lab is and has been release testing based on the P/E-ABI,
> PTC, derivitization and subsequent separation on c-18 R/P chromatography
> system. As I perceive the story, this instrumentation is no longer being
> supported and what little support there is, now, will eventually disappear
> all together.
> So to make a long story short, we now have in our laboratory, a
> system which has Beckman's name on it, to be exact; 126AA solvent module,
> 166 detector, 232 post column reactor, 507 system gold autosampler and a 350
> IBM Personal computer. I have been validating this system and find that it
> works acceptably well, but the autosampler is something from a different
> realm. I understand that Beckman makes a 508 autosampler which does not have
> a column oven incorporated into it's design. Any experience with this
> situation would be appreciated. Should I up-grade to the 508 and buy a
> column oven, which by the way would have to be programmable to accommodate
> the chromatography or should I struggle along with the current autosampler
> which uses three times the actual amount of sample needed for injection.
> Another option would be to switch the whole post column ninhydrin system
> over to an HP1100 or a Waters Alliance system any and all constructive
> comments would be appreciated.
>
> James G. Freedy
> Ortho Biotech
> Raritan, N.J.
>
>
>