HPLC for formulation

Hi everyone!

My name is Azmar. I am a MSc student and my research involves plant extraction, compound isolation and formulation. I used my isolated compound as my main ingredient in my formulation. To know the content of the isolated compound incorporated in the formulation, I use high-performance liquid chromatography (HPLC) instrument to detect and quantify the isolated compound.

My question here is do I also need to run HPLC for all ingredients that I used for the formulation?

The reason of running HPLC for all ingredients is because there might be overlap in chromatogram of other ingredient with my isolated compound. But I know the isolated compound has its own retention time and peak purity.

So right now I am kind of confuse whether to run HPLC for other ingredients as well.
:-S

Can anybody help me on this? Thank you in advance.

Comments

  • What is your formulation? And why would you want to run HPLC on the rest of the ingredients? What do you want to find?

    As I know, HPLC separates a fairly 'clean' mixture into its components and quantifies those components based on a single factor. So you run your plant extract through and look for, say, menthol. Is that correct? Now the rest of your ingredients, either don't contain menthol, correct? So what would the HPLC detect?
  • DavidDavid Member
    edited June 2015
    Exactly - to be able answer we need to know what are you looking for and why! :)
  • RubenRuben Member
    Assuming your have the right column, mobile phase, detector, method, etc this is what I would do.

    I would prepare one formula with your compound and one without your compound, run through the HPLC and then overlay the chromatograms.

    if you see that in the chromatogram without your compound nothing is eluting at the time your compound elutes then you are fine.

     
  • Mike_MMike_M Member
    My question would be what are you trying to measure, and what kind of detector are you using? Not every ingredient is going to be picked up by the detector depending on what you've chosen.

    If you're just trying to show your formulation with the ingredient compared to a background without it to illustrate your active ingredient is indeed creating the peak then I would follow the above instructions.
  • Dear AuroraBorealis, David, Ruben and Mike_M,

    Thank you so much for your replies!

    My isolated compound is a phenol. I get your point there, AuroraBorealis. This means HPLC is used to detect a particular compound based on the specified parameters (column, mobile phase etc). Running through all ingredients blindly using the same parameters might give noise or no result and of course, HPLC won't detect my isolated compound because it isn't there.

    Ruben, your suggestion make sense and it's practical. I'll see what I can do.

    Thank you so much!
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