Hi,
Long story short, the HPLC didn't listen to the method batch and continued to run dry after pumping half a liter of water through it. It will have been sitting there for 5-6 days dry. Is there any way of saving the column?
Column is a Superdex 200 increase
Drying can lead to the collapse or irreversible alteration of the gel matrix, making the column ineffective or degraded.
Youc an try to rehydrate and possibly save the column. After you can check your SEC column with calibration kit :
https://www.cytivalifesciences.com/en/us/shop/chromatography/prepacked-columns/size-exclusion/gel-filtration-calibration-kits-lmw-hmw-p-05801?srsltid=AfmBOoq0qtXmiMU2ltAgVClB5KL-9sw0YCZ_kQpCBliQrJ3HOk8aY5hW
First, flush the column with the mobile phase (water or the buffer you typically use in your method) at a low flow rate (around 0.5-1 mL/min or lower, depending on the column specifications).
Gradually increase the flow rate to prevent any sudden stress on the stationary phase. Start with a lower flow rate (like 0.5 mL/min) and gradually increase to the intended flow rate after confirming there's no leakage or pressure issues.
If you suspect the column has dried out more extensively, a stepwise buffer exchange could help. Start by flushing the column with low-concentration buffer (e.g., a diluted version of your regular buffer) and gradually increase the concentration over a few hours, allowing the column to rehydrate slowly.
Keep a close eye on the pressure (and Deltapressure if possible) as you rehydrate. If the pressure spikes significantly or if the flow rate is difficult to maintain, the stationary phase might have collapsed or degraded.
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