Sample not eluting on different system

Hi folks,

Currently losing my mind over the following situation:

We got a brand new semi-preparative system on which I want to run a chiral separation.

On the analytical instrument I get the chromatogram I expect, using an isocratic method.

On the semi-prep I use the same (as in physically the same) column and sample solution and don't get any peaks at the respective wavelength. Extended runtime to 3x the expected length and just get baseline response.

Details on the systems: Analytical Shimadzu CBM40 controller LC40D pump SIL 20A HT autosampler SPD M40 detector (PDA)

Semiprep Shimadzu CBM40 controller LC20 AP pump SIL 10AP autosampler (5mL loop) SPD 40 detector (dual wavelength)

Column used is a Chiralpak IB3 250x4.6mm running isocratic H2O/MeCN 40:60 @ 0.8 mL/min

Mobile phase is mixed by the instruments. Sample injection tried at 10, 50 and 100 microliters (concentration is about 1 mg/mL) Flow cell is identical on both instruments

So far I have tried: -Replace MeCN and H2O bottles

-premix mobile phase instead of relying on the instrument

-replace rinse solution (same as mobile phase) for sample loop

-purging column with 100% MeCN and reequillibrating

-increasing %B during run

-Check solvent delivery accuracy of semiprep (both flow and ratio)

-performed sanity test by using a C18 column and caffeine (50mg/L) in the same general setup (this worked)

I'm running out of ideas and the senior scientist I consulted is also baffled by this, any ideas and suggestions are welcome.