Protein containing Polysorbate 80 and RP HPLC
Posted: Wed Mar 29, 2006 2:04 pm
Hi!
Recently Polysorbate 80 has been added to the manufacturing process. I am running on a VYDAC C18 MS Column Cat # 218NS54 5um and the mobile phase is 25% Methanol, 50% Water and 25% ACN. I diluted the sample,which is in 0.01% Tween 80 in 10 mM Citrate, 100 mM Glycine and 100 mM NaCl, down to 5.0 mg/mL. My problem is that I am seeing a shift in the retention time. My question is would a wash with 90% ACN and 10% water for 10 minutes each injection help my retention time? Is there another way to ensure that the retention time will stop shifting? I would really appreciate the help! Thanks
Recently Polysorbate 80 has been added to the manufacturing process. I am running on a VYDAC C18 MS Column Cat # 218NS54 5um and the mobile phase is 25% Methanol, 50% Water and 25% ACN. I diluted the sample,which is in 0.01% Tween 80 in 10 mM Citrate, 100 mM Glycine and 100 mM NaCl, down to 5.0 mg/mL. My problem is that I am seeing a shift in the retention time. My question is would a wash with 90% ACN and 10% water for 10 minutes each injection help my retention time? Is there another way to ensure that the retention time will stop shifting? I would really appreciate the help! Thanks