TylerSmith123 wrote:
Regardless of the methodology, is there a way to switch your mobile phase component? I'd be very worried that precipitation will occur in the bottle (especially if the temperature fluctuates in your lab), or onto the column you are using. I assume this is apart of a binary phase where this is one of your reservoirs, while the other is the aqueous portion. Could the phases be juggled a little better by any chance? For example, if you're using a 100% aqueous MPA (mobile-phase A), and this mixture as your MPB, then could it be possible to get at least a similar separation with a MPA that includes 10% ACN and a MPB with a concentration of acetonitrile less than 95, maybe like 75-85%?
Perhaps I'm just being pedantic but this would have me very worried when it comes to potential precipitation.
Thanks for advice I think it will be good to make sure to get some ACN mixed into the water phase really well as you suggest. I was using a 2 L bottle that did not fit into the sonication bath so I probably added to much ACN before getting a the first addition mixed enough. This resulted in a bottom layer with H20 and AmAc but it dissolved over night. The method is quite complicated running like 200 MRMs so I have speed a lot of time with validation which is really time consuming to get all compounds right. I am very careful with the ammonium acetate and never leave it the system without flow and have not had any problems so far.
Thanks for the advice and some options to test if problem persists.
Per