We continue to have problems with peak tailing even after: an entire system clean-up including H2O/Isopropanol/Nitric Acid/NaOH, trial of 3 different columns (Perkin Elmer, 33 x 4.6mm, C18, 3um RP)(all from different lots), changed the in-line filter, checked the pH of buffer (3.4) and filtered HPLC grade ACN is the mobile phase.
Any and all suggestions for a remedy would greatly be appreciated.

What compound you are interest in? You may need to change mobile phase. Have you observed a nice peak with this PK column under current chtomatographic condition?

If you suspect the system, measure the peak width without the column (preferentially with your sample and mobile phase) and compare it too the peak width with the column.
From a chromatogram with many different compounds in it eluting at different retention times, you can judge, whether the problem is a column problem or a system problem. If the peaks get better with increased retention, one may suspect the system. If the peaks are all lousy, independent of retention, you got lousy columns. If something in between is happening, we need to understand the details.

Thank you for your responses. I guess I should have given more information.... we are analyzing dansylated polyamines and have been doing so in our lab for many years using the same solvents and gradient profile (with nice separation and peak shape). This is a sudden problem and equally effects all 4 of our peaks of interest, even in our standards. After an intense system cleaning the peak shape was only a bit sharper as it was with using a different lot of column, but there was still tailing and the shape was not great... if it is a systemic problem what are the likely suspects? Thanks again for all of your input.

One possible solution is to remake all your fittings. One improper ferrule or fitting with a little deadspace can cause the phenomenon you're seeing, especially post-column.

Any recent changes to the system? Someone inadvertently using large ID tubing could cause tailing.

It is still worth to measure the system bandspreading, or visually check if the tubing to the detector is not a pipeline. Sometimes the inlet-oulet lines get confused, and the detector outlet are always pipelines....

I think it might be productive to try to reproduce a different separation. For example, try to reproduce a simple separation that is used for calibration or OQ/PV testing. If this separation is also affected similarly you can probably rule out column/mobile phase chemistry and concentrate on the system.

I feel Uwe's comments are valuable, probably even more so if you had run them as a benchmark when the system was known to have been operating properly. Simple benchmarks like a bandspread and dwell volume tests can be run in a few minutes in conjunction with annual/semiannual calibrations and save a lot of time when system troubleshooting.

Good luck and let us know what the problem was.