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- Posts: 2
- Joined: Mon Jan 21, 2019 8:23 am
I am currently working with a Microsorb 100-5 NH2 250 x 4.6 mm column, trying to separate barley beta-glucan hydrolysates products (e.g. glucose, cellobiose, and other oligosaccharides DP<7). Unfortunately, after the first successful separation, when I run my standards I saw extra peaks in the trisaccahride and tetrasaccharide chromatographs and the backpressure was very high. What I did was to initially wash the column with multiple volumes of the mobile phase (70:30 ACN:water), both the right way and the reverseof the column, then with pure ACN, then with a gradient of 50:50 MeOH:water to 100% MeOH but still the pressure is very high. Namely, the backpressure is currently 200 bar at a flow rate of 0.5 mL/min. I am currently washing the column with 50:50 MeOH :Chorophorm as suggested in the booklet of the column. Would you have anything else to suggest? Thank you in advance.