UPLC Analysis of B vitamins

<p> Hi Everyone :</p><p></p><p>I'm looking to see if there are other users who have developed surefire methods or know of such methods that already exist for the analysis of a wide range of B vitamins by UPLC.</p><p>Here's what I need to separate and quantify . . .</p><p></p><p>Vitamin B12</p><p>Calcium Pantothenate [B5]</p><p>Niacinamide [B3]</p><p>Pyroxidine HCL [B6]</p><p>Thiamine HCL [B1]</p><p>Biotin [B7]</p><p>Folic Acid [B9]</p><p>Riboflavin [B2]</p><p></p><p>I know of the Water's UPLC BEH Amide column, but just how good is it for this particular set of B vitamins ?</p><p>Curious to know before I invest.</p><p></p><p>Thankyou all</p><p>p</p>


  • lizh


    With vitamins, the standards can work very well, the trick is always the matrix, so that being said,  where are the samples from and what sensitivity is required!

    Good luck,


  • Hi,

    in our lab we developed an UPLC-MS/MS method for the quantification of folic acid and key folate forms in serum or plasma samples.

    The work was published in Kirsch, SH et al. J Chrom B 2010, 878(1);68-75

    We used an HSS T3 column for the separation.

    I hope this is of interest and can help you.



  • rune

    Hi Bartelby

    You can find most of the compounds on the last page in the attached application notebook for the BEH Amide column.

    best regards


  • Thankyou all very much for your responses !!  Much appreciated.

    I'll be checking these vitamins in cell culture media.  I don't know just yet, but I may have to inject the media straight without dilution in order to detect them.  I'll do all the proper analytical work and see how it goes, but it looks like there are a couple good Water's columns to do the job.

    Thanks again everyone !!

    Have a great weekend.


  • Hello,

    Do you have an idea as to what separation mode you are planning (HILIC vs reversed phase)? If using HILIC, the cell culture media is likely 100% aqueous and that is too strong an injection solvent - you'd need to dilute with some organic modifier to more match the ACN-rich HILIC mobile phase(s).


  • Hey Doug, thanks.

    Yah, I've got a couple methods I'm going to play around with, both that allow aqueous starting conditions.

    Darn, I wish I had a Mass Spec ; would make life a lot easier on this analysis.  Maybe in 2011.