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- Posts: 4
- Joined: Sat Sep 10, 2016 5:04 pm
Every time before changing the buffer, I flushed the column thoroughly with water and 30%ACN, it shouldn't be the retention problem (the retention times are very reproducible under each mobile phase). I also noticed that even the calibration curve made with polystyrene sulfonate are not linear at the low Mw, the slope became less steep, which also confuse me.
I wonder if acetone is a good marker for the exclusion volume, and how can I determine the exclusion volume at its optimal condition (all the secondary interactions are suppressed to the minimum)? More importantly, can anyone tell me why the calibration curve became less steep at the lower Mw?
Many thanks in advance!