You may wash pumps and the column with 5% Phosphoric acid (in A and B
positions). Purge, then wash for 1 hour (325 ul /min or your usual flow
rate), wash 15 min with water and re-equilibrate with A and B.
This helped us recently with a similar problem on our Procise.
Have you checked your dynamic mixer on the HPLC?
Good luck
Henriette
>Dear ABRF:ers
>I am running a 494 Procise Sequencer and I have trouble with
>the PTH separation. Please have a look at the attached chromatogram.
>I would bee very happy if somebody could figure out what's wrong.
>I have changed all the pump seals. Solvents A , B and Premix tested.
>New column and precolumn installed. The chromatograms look the same through
>the whole testing procedure; a big slope in the beginning and the
>hydrophobic PTHs elute too early.I am getting desperate.
>
>Thanks in advance.
>
>Attachment converted: +-:Standard.JPG (JPEG/JVWR) (0000B880)
>Christer Wernstedt
>
>
>Ludwig Institute for Cancer Research
>Box 595
>S-75124 Uppsala
>Sweden
>
>Phone: +46-18-160424
>Fax: +46-18-160420
>E-mail christer.wernstedt@licr.uu.se
>Home page http://www.licr.uu.se
Henriette A. Remmer, Ph.D.
Director of Protein Services
Protein Sciences Facility at the Biotechnology Center
University of IIllinois
315/311 Noyes Laboratory, Box 62-1
505 S Mathews Ave Urbana, IL 61801
phone: (217)333-4695 or (217) 333-3841
fax: (217)244-1142 email: remmer@uiuc.edu
homepage: http://www.life.uiuc.edu/biotech/protein_sc.html
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