Chromatography Forum

Dear All,
I am trying to find a information about analysis of Folic Acid on foods, I would like to find a good procedure, I can Use UV or fl deteccion, my other question is:
Need I a derivatation step?

Thanks!

Folic acid can be separated on our Cadenza CD-C18 column:

http://www.silvertonesciences.com/modul ... TI060E.pdf

Hamilton application note #428 (I can send you the PDF file if you contact me).

Summary of conditions:
PRP-1 column (79422, 4.1x250mm)
1ml/min
MP: 2.5% ACN, 97.5% 20mmol/L sodium phosphate buffer pH=6.2
T= ambient
UV at 280nm

best regards

e-mail: rhaefele@hamiltoncompany.com

Bryan, the following question is not only directed at you, it´s just that I now felt compelled to ask it: Separated from what?

HW -

(As foot is taken out of mouth) - perhaps "analyzed" as opposed to "separated" would have been more appropriate for this chromatogram.

Good stuff!

How about: "shows retention on" ......? That way you can´t be held accountable for a lousy separation.

Separation OF folates IN plasma was studied ON different columns.

Reversed phase and several polar columns were tested. However, the final choice was HILIC on a poly(2-hydroxyethyl aspartamide) silica column.

Garbis, S. D, Melse-Boonstra, A, West, C. E, van Breemen, R. B,
Anal Chem 2001, 73, 5358-5364

for chromatographic condition, you may refer to USP monograph for folic acid. it uses UV detection.

I recalled a related substance called folinic acid. I think USP requires that folinic acid and folic acid be separated with a resolution of > 2, as a suitability requirement.

Dear All,
I am trying to find a information about analysis of Folic Acid on foods, I would like to find a good procedure, I can Use UV or fl deteccion, my other question is:
Need I a derivatation step?
Thanks!

You have to decide what you want to measure. If you want to measure the total of both naturally-occuring and added supplement, you may have to perform a derivatisation. If you want to measure just one type, then you probably can extract and analyse. The methods in pharmacopoeia are for formulated pharmaceuticals.

As you want to measure in foods, I'd look to publications like the AOAC and Food Chemicals Codex for methods and current perceptions, There were concerns at discrepancies between microbiological data and HPLC data for folate intake, but they may have been resolved now.

I recommend the book below for anyone working on HPLC analysis of vitamins. The articles below might also provide some data.

A validated liquid chromatographic method for determining folates in vegetables, milk powder, liver and flour.
Konings EJM.
J AOAC Int 1999 vol.82 p.119–27

Water-soluble vitamins.
Konings EJ;
J AOAC Int. 2006 Jan-Feb;89(1):285-8.

Modern chromatographic analysis of vitamins. 3rd ed.
de Leenheer AP, Lambert W, and Van Bocxlaer JF, eds.
New York: Marcel Dekker, Inc, 2000 p.301–23.