Size exclusion peptide with AdvanceBio SEC, 130Å, 2,7 µm, 4,6 x 300 with PL1580-5350 part number

Question

Hello, 
 We have HPLC Column AdvanceBio SEC, 130&Aring;, 2,7 &micro;m, 4,6 x 300 with PL1580-5350 part number. We use this column to analyse the peptide product the running method is as follows: K2HPO4 20mM, pH 6.8 Flow rate: 0.5ml/min Column temp: 25C Detection wavelength: 220 nm (UV) Injection volume 5 ul The starting pressure of the system with the guard was 175 bar. After 30 samples running, the column pressure increased to 200 bar and the peak resolution was decreased. After watching this problem we washed the column according to the column user manual with Na2SO4 1M with pH 3 but the problem remained. Please help us resolve this problem. 
 Tnx

jens · Accepted Answer

Hi, 
 I passed this query on to a colleague who was involved with developing these columns, this was their response: 
 
 "It might be worth checking if they are filtering their mobile-phase and samples as it's possible the guard may be fouled. They could check it&rsquo;s the guard they are affecting by doing a single run without the guard to see if performance returns." 
 Hope this helps

Size exclusion peptide with AdvanceBio SEC, 130Å, 2,7 µm, 4,6 x 300 with PL1580-5350 part number

Top Replies