UPLC BEH Amide = USP L60 ?

sazimi
sazimi
in Instrumentation and Mass Spectrometry
<p>Dear UPLC users & friends</p><p></p><p>I need to develop & transfer some methods on UPLC by USP L60 column (alkyl amide groups & endcapped) previously worked with Ascentis RP-Amide & Polaris Amide-C18 on Breeze & Alliance so I know very well about the beauty of alkyl amide columns, I just see Waters recently launched UPLC BEH Amide columns....... I request all Waters & UPLC specialists please confirm BEH Amide is in USP L60 listing or not..?. If anyone have applications, seminar or presentation on UPLC BEH Amide pls attach with your reply </p><p></p><p>Thank you.</p><p></p><p>Salman Azimi </p>

Answers

  • dougm
    dougm

    The USP L60 designation is for a reversed phase column chemistry that has an embedded amide polar group. The closest chemistry that we have to that is our BEH Shield RP18 chemistry. The BEH Shield RP18 chemistry is listed as an L1 column but contains a carbamate embedded polar group.

    The BEH Amide column is just the amide chemistry with no endcapping. It is a column designed for HILIC separations, not reversed phase, and as such is not a USP L60 chemistry. In fact, to our knowledge, there presently is no USP L classification for the BEH Amide chemistry.

    --Doug

  • sazimi
    sazimi

    Dear Doug

    Thanks for your comments, you mean BEH Amide is normal phase column in HILIC mode & bonded with amide group if so it should be good column for hydrophilic compounds like polyalcohols, oligosaccharides, carbohydrates etc. but due to HILIC mode it must require high composition of ACN or MeOH.

    As I experienced HILIC columns are good for highly polar compounds with better sensitivity & early retention in LC-MS compared to reversed phase, but elution of polar compounds in natural products always difficult in HILIC, what I understand in BEH Amide due to Amide linkage in HILIC mode this column might help me to get better separation of polar compounds in natural & dietary products, Isn't it?

    If so please confirm more about the application of BEH Amide, that I am looking to order this column for my LC-MSMS analysis in future. If you have any details, application about BEH Amide pls attach.

    Salman Azimi

  • Benzene
    Benzene

    I have an additional question related to BEH Amide column.

    I would like to know maximum how much I can use aqueous phase on this column. I checked applications on Web, so I know it is unnecessary to use much aqueous phase on this column. I just want to give co-workers' a head-up about usage when I get this column.

    Best Regards,

  • sazimi
    sazimi

    According to Doug, Senior Marketing Manager, Waters; BEH Amide is normal phase column in HILIC mode with covalently bonded amide groups, so it is not USP L60 packing, because in this listing most columns are RP, endcapped, amide linked with highly aqueous phase compatible.

    In HILIC mode, you need high composition organic mobile phase but for highly polar small drug molecules you can minimize organic to 45-50%.

    Due to amide in HILIC mode, this column should be best for polyalcohols like saccharides, OH interaction with amide but in this case, you need 70-80% organic, & off-course analysis also depend on your compounds chemistry.

    You can use slightly high aqueous mobile phase for peptides with BEH Amide.

    Better answer will give you Waters peoples but my view this column is specialized for polyalcohols, sugars (as in ion-exchange columns) or amino acids, in high organic mobile phases. If you are looking for highly aqueous mobile phases dont go for this column better to select BEH Shield or HSS T3

    I'm also waiting for more clarification !!!! that I used last year in 2-3 samples "TSK gel Amide-80 HILIC", same chemistry like BEH Amide only BEH difference, but TSK gel Amide-80 columns are in USP L60 listing. but Mr. Doug saying BEH Amide has no USP listing? Its better to get satisfactory answer before ordering.

    Salman Azimi

    Message was edited by: Salman Azimi

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