AOAC SPIFAN ERP Meeting Book (3-16-16)

Contribution of minor folates to the total folate content of infant formula and

adult/pediatric nutritionals / F. Martin (NRC, AS)

03 Feb 2016

CONFIDENTIAL

ͼ This document may not be reproduced or disclosed to third parties without prior authorization

Page 2 / 13

B A C K G R O U N D A N D O B J E C T I V E S

The Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) of the AOAC has identified

vitamin B

as one of the nutritional parameters which needs new analytical methodology for its

determination. The Expert Review Panel (ERP) reviewed and granted First Action status to two

methods: AOAC Official Method 2013.13 (Nestlé’s method) and AOAC Official Method 2011.06

(Silliker’s method). The two methods differ on several points, which are summarized in the following

table:

Parameter

AOAC 2013.13

AOAC 2011.06

Compounds targeted

Folic acid, 5ͲmethylͲ

tetrahydrofolate

Folic acid, 5ͲmethylͲtetrahydrofolate,

5ͲformylͲtetrahydrofolate,

tetrahydrofolate, 5,10 methenylͲ

tetrahydrofolate, 10ͲmethylͲfolic

acid, 10ͲformylͲfolic acid

Extraction procedure

DualͲenzyme (amylase,

protease)

Trienzyme (amylase, protease,

deconjugase)

SPE cleanͲup

Strong anion exchange (SAX)

Weak anion exchange (WAX)

It is worth noting that the SMPR (Standard Method Performance Requirements), issued by the

SPIFAN, target folic acid and 5ͲmethylͲtetrahydrofolate (including the 5ͲmethylͲtetrahydrofolate

polyglutamates).

During method development of AOAC 2013.13, experiments showed that other minor folates

potentially present in infant formula and adult nutritionals represented a minor contribution to the

total vitamin B

content. Moreover, trials with deconjugase showed only a minor liberation of

polyglutamate folates.

The main objective of this note is to assess the content of minor folates

in products belonging to the

SPIFAN II kit with and without inclusion of the polyglutamate forms.

Minor folates targeted are: 5ͲmethylͲtetrahydrofolate (5ͲMe THF), 5,10 methenylͲtetrahydrofolate (5Ͳ

10MylͲTHF), tetrahydrofolate (THF), 5ͲformylͲtetrahydrofolate (5CHOͲTHF), 10ͲformylͲfolic acid (10CHOͲFA)

and 10ͲmethylͲfolic acid (10MeͲFA).

2013.13 (Fol-21) / (February 2016) - NOTES

FOR ERP USE ONLY

DO NOT DISTRIBUTE