Dear all,

I am new with chromatography, I put my hand on old Beckman coulter 126 equipped with 166 detector UV / VIS. The instrument was not used for Approximately 12 years and now I am starting to check before to start my new research project.

To test I have started to clean everything without column and I flush for several times at high flux rate (10ml / min) Mobile phase (H2O: CH3COOH and CH3COH: CH3COOH 99: 1 everyones), previously degassed via ultrasound bath.

After this operation I plugged my column, an other old pieces, (phenomex moon 5u C18 250mm), after some problems I was able to eluate my sample test (Anthracene in Methanol) at the same time.

So I started to check the integrals under the peak for different Concentrations of my sample. Unfortunately I have problem, I am not able to build a calibration curve, the behavior of integrals is not linear and not reproducible.

Then I started to check every things, and I observed that Rheodyne system has some troubles, when i put my sample, some drops go out as it is obstruited. I have also observed that any drops flow out from the waste capillar when I put quantity of solution more than 20microliter.

What can I do? I am thinking to disassemble Rheodyne system and check, but I am not so very handy if you have tutorials about disassembling of Rheodyne, some tips or some alternative solution, It could be very useful!

Thank you

Valentino Russo

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