HPLC Separation Modes

Chromatographic ModeAbrv.Column TypeMobile PhaseApplication
Reverse-phase chromatographyRPCNon-polar (e.g. C18)Polar mixture of water and organic solvent (e.g. acetonitrile)Water-soluble samples
Normal-phase chromatographyNPCPolar ( e.g. unbonded silica)Less-polar (than stationary phase) mixture of organic solvents (e.g. hexane, ethyl ether, chloroform, methylene chloride) Water-insoluble samples, isomer separation, and preparative HPLC
Non-aqueous reverse-phase chromatographyNARPNon-polar (e.g. C18)Mixture of organic solvents (e.g. ACN + methylene chloride)Very hydrophobic samples
Hydrophilic interaction chromatographyHILICPolar (e.g. silica or amide-bonded)Mixture of water organic solvents (e.g. ACN + H2OHighly polar samples that are poorly retained by reverse-phase mode
Ion-exchange chromatographyIECUsually an organic resin that has charged groups able to bind ions of opposite chargeAqueous solution of a salt with buffer.Separating ionizable samples, large biomolecules (e.g. proteins, carbohydrates)
Ion-pair chromatographyIPCNon-polar (e.g. C18)Polar mixture of water and organic solvent (e.g. ACN) w/ an ion-pairing reagent (e.g. alkylsulfonates, trifluoroacetic acid) that interacts w/ sample ions of oposite chargeAcids and bases that are weekly retained by reverse-phase
Size-exclusion chromatographySECInertAqueous or organicLarge biomolecules, polymers. Separates by molecular weight.

10 Responses to “HPLC Separation Modes”

  1. It’s difficult to find well-informed people on this subject, but you
    sound like you know what you’re talking about! Thanks

  2. We are grateful to be separated by models, could find necessary.

  3. match.com says:

    I’m amazed, I must say. Rarely do I encounter a blog that’s both educative and amusing, and without a doubt,
    you’ve hit the nail on the head. The issue is something
    which not enough men and women are speaking intelligently about.
    I am very happy I stumbled across this in my
    search for something relating to this.

  4. Zum Artikel says:

    Hey it’s a nice post you got here and I enjoyed reading your article.I hope to find some nice news here. And of course I bookmarked it, Thank you for sharing!!

  5. reformas says:

    I like what you guys tend to be up too. This kind of clever work and coverage!

    Keep up the very good works guys I’ve incorporated you
    guys to my blogroll.

  6. thanks for inspiring me ….

  7. Weiter says:

    Hi there, just became aware of your blog through Google, and found that
    it’s truly informative. I’m going to watch out for brussels.
    I’ll be grateful if you continue this in future. Lots of people will be benefited from your writing.

  8. Homepage says:

    Thanks for a marvelous posting! I really enjoyed reading
    it, you could be a great author. I will make sure to bookmark your
    blog and will eventually come back later on. I want to encourage you to definitely
    continue your great writing, have a nice afternoon!

  9. Thank you for some other great article. The place else could
    anybody get that kind of information in such an ideal method of
    writing? I have a presentation next week, and I’m at the search for such information.

  10. I’m not sure where you’re getting your info, but good topic.
    I needs to spend some time learning more or understanding more.
    Thanks for magnificent info I was looking for this info for my mission.

Leave a Reply

Recent Entries

    Recent Comments

      Most Commented