Chromatography Forum

Hello, I have to develop a UPLC and a UHPLC method for a new active product soon- assay and impurities and although im very experienced with chromatography, its more from the end-user point of view ie using validated methods, running and processing samples etc. I don't know where to start with choosing the right column, isocratic or gradient, best solvent etc, are there any good guides out there to jump into so at least I have some kind of background to it and start creating new methods, test Resolution, sensitivity, peak shape etc?

Also, obvious question here but is it better to run a fixed concentration of standard and run a sample to confirm you are recovering the total amount? How do you know that your peak areas etc match up to what you injected? Thanks.

Our hosts (Separation Science) have a series of self-paced video courses that are quite reasonable priced. Check out:
https://www.analytical-training-solutio ... by-design/ taught by John Dolan

By way of "full disclosure", I've prepared some of the courses for them.

Thanks very much Tom, I will look into those courses as I need a foundation before I can start developing methods.

John W. Dolan: The Perfect Method
Article series in LC-GC (2007)
http://www.chromatographyonline.com/sea ... 20Dolan%22

If you share composition and structure of your compounds I can tell you exactly what to do. You can also send me a sample for free method development screening. If NDA/CDA is required we can sign it. We usually have a method in one day.

Thanks very much for the offers of help. I don't know the exact nature of the molecule as of yet but ill check back in when I do.

I don't need to know the exact structure only if it is hydrophobic or hydrophilic and if it is neutral, basic or acidic. here is a little bit of explanation how exact structure does not matter in method development:
https://www.linkedin.com/feed/update/ur ... 6767311873

Thanks Vlad, at this stage I do know the molecular weight and what kind of compounds they are, its three compounds in one product. Im looking to develop an assay method using reversed phase chromatography on a Waters UPLC or H-Class instrument, would you still be able to help with the starting guidelines for this?

Sure. Please send me details by email (through our website)