I would like to measure the Vitamin B5 content in the raw material. Can anyone tell me how to measure vitamin B5 by using the HPLC-UV and which column should we choose?

Hi Yangbei,

yes, those columns would fit into an Agilent LC instrument. You need to check the fitting of the column against the ones of your connection tubing on the Agilent machine, sometimes another ferrule/fitting needs to be used. There is a webinar about this here.

But have you had a look at our two application notes about multi-vitamin samples?

• Simultaneous Detection and Quantitation of 14 Water-Soluble Vitamins in a Supplement by LC/MS/MS Triple-Quadrupole
• Analysis of water-soluble vitamins from multivitamin tablets for nutrition labeling

Both done on Poroshell columns and very fast, Vitamin B5 eluting between 3.5 and 3.7 minutes.

Regards,

Norbert

Hi Dr. Norbet

Thanks for your reply. But I do have a question about the second paper you refer to me (Analysis of water-soluble vitamins from multivitamin tablets for nutrition labeling). I can't find the sample preparation method. In that paper, it uses the multivitamin tablet so that the vitamin content can fully dissolve into the water. What about the sample with hard texture? Do I need to use acid to break the protein and then fully extract the vitamin B5 in the sample? 

Hi yangbei,

the sample preparation is described on page 3 under the headline "Sample Preparation", the tablets are dissolved in 200 mL water under sonication with an added amount of 0.25M-NaOH and the solution is filtered through a syringe filter with 0.25 µm pore size.

If your sample is harder you have to finely grind it first, put it in the water and sonicate longer.

Regards,

Norbert

Hi yangbei,

the sample preparation is described on page 3 under the headline "Sample Preparation", the tablets are dissolved in 200 mL water under sonication with an added amount of 0.25M-NaOH and the solution is filtered through a syringe filter with 0.25 µm pore size.

If your sample is harder you have to finely grind it first, put it in the water and sonicate longer.

Regards,

Norbert

Hi Dr. Norbet 

Thank you very much